New England Biolabs Canada
 

Product Pathways - Antibody Sampler Kits

Human T Cell Co-inhibitory and Co-stimulatory Receptor IHC Antibody Sampler Kit #44689

Item# Description List Price Web Price Qty
44689T Human T Cell Co-inhibitory and Co-stimulatory Receptor IHC Antibody Sampler Kit - 1 Kit $808.00
$727.20
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
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Companion Products

Item# Description List Price Web Price Qty
12630P SignalFire™ Plus ECL Reagent - 20 ml $116.00
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12630S SignalFire™ Plus ECL Reagent - 100 ml $519.00
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12757P SignalFire™ Elite ECL Reagent - 20 ml $123.00
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12757S SignalFire™ Elite ECL Reagent - 100 ml $558.00
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12957S Western Blotting Application Solutions Kit - 1 Kit $599.00
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13953L Prestained Protein Marker, Broad Range (11-190 kDa) - 1750 µl $496.00
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13953P Prestained Protein Marker, Broad Range (11-190 kDa) - 50 µl $60.00
$54.00
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13953S Prestained Protein Marker, Broad Range (11-190 kDa) - 350 µl $122.00
$109.80
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14058S B7-H3 (D9M2L) XP® Rabbit mAb - 100 µl $433.00
$389.70
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14058T B7-H3 (D9M2L) XP® Rabbit mAb - 20 µl $184.00
$165.60
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14746P SignalStain® Citrate Unmasking Solution (10X) - 25 ml $36.00
$32.40
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14746S SignalStain® Citrate Unmasking Solution (10X) - 125 ml $133.00
$119.70
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15019L Animal-Free Blocking Solution (5X) - 250 ml $193.00
$173.70
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15019S Animal-Free Blocking Solution (5X) - 25 ml $57.00
$51.30
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15372S LAG3 (D2G4O™) XP® Rabbit mAb - 100 µl $433.00
$389.70
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15372T LAG3 (D2G4O™) XP® Rabbit mAb - 20 µl $184.00
$165.60
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34594S 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb - 100 µl $385.00
$346.50
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45208S TIM-3 (D5D5R™) XP® Rabbit mAb - 100 µl $609.00
$548.10
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45208T TIM-3 (D5D5R™) XP® Rabbit mAb - 20 µl $207.00
$186.30
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5425S Normal Goat Serum - 10 ml $92.00
$82.80
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54979S VISTA (D5L5T) XP® Rabbit mAb - 100 µl $433.00
$389.70
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54979T VISTA (D5L5T) XP® Rabbit mAb - 20 µl $184.00
$165.60
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61637S OX40 (E9U7O) XP® Rabbit mAb - 100 µl $433.00
$389.70
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61637T OX40 (E9U7O) XP® Rabbit mAb - 20 µl $184.00
$165.60
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68014S GITR (D9I9D) Rabbit mAb (IHC Preferred) - 100 µl $385.00
$346.50
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68014T GITR (D9I9D) Rabbit mAb (IHC Preferred) - 20 µl $164.00
$147.60
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6883P3 SignalFire™ ECL Reagent - 50 ml $43.00
$38.70
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6883S SignalFire™ ECL Reagent - 500 ml $370.00
$333.00
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7074P2 Anti-rabbit IgG, HRP-linked Antibody - 100 µl $109.00
$98.10
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7074S Anti-rabbit IgG, HRP-linked Antibody - 1 ml $203.00
$182.70
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7074V Anti-rabbit IgG, HRP-linked Antibody - 5 ml $646.00
$581.40
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7727L Biotinylated Protein Ladder Detection Pack - 3250 µl $326.00
$293.40
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7727S Biotinylated Protein Ladder Detection Pack - 650 µl $92.00
$82.80
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8059P SignalStain® DAB Substrate Kit - 1 Kit $60.00
$54.00
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8059S SignalStain® DAB Substrate Kit - 1 Kit $200.00
$180.00
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8112L SignalStain® Antibody Diluent - 100 ml $117.00
$105.30
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8112S SignalStain® Antibody Diluent - 25 ml $57.00
$51.30
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8114P SignalStain® Boost IHC Detection Reagent (HRP, Rabbit) - 1 ml $56.00
$50.40
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8114S SignalStain® Boost IHC Detection Reagent (HRP, Rabbit) - 15 ml $525.00
$472.50
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81254S PVR/CD155 (D8A5G) Rabbit mAb - 100 µl $385.00
$346.50
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86163S PD-1 (D4W2J) XP® Rabbit mAb - 100 µl $433.00
$389.70
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86163T PD-1 (D4W2J) XP® Rabbit mAb - 20 µl $184.00
$165.60
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9997S Tris Buffered Saline with Tween® 20 (TBST-10X) - 1000 ml $90.00
$81.00
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9998S BSA - 50 g $313.00
$281.70
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9999S Nonfat Dry Milk - 250 g $86.00
$77.40
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VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
PD-1 (D4W2J) XP® Rabbit mAb #86163 20 µl W, IP, IHC-Bond, IHC-P, F H 52-65 Rabbit
TIM-3 (D5D5R™) XP® Rabbit mAb #45208 20 µl W, IP, IHC-Bond, IHC-P, F H 45-70 Rabbit IgG
LAG3 (D2G4O™) XP® Rabbit mAb #15372 20 µl W, IHC-Bond, IHC-P H 60-80 Rabbit IgG
VISTA (D1L2G™) XP® Rabbit mAb #64953 20 µl W, IHC-Bond, IHC-P, F H, Mk 45-65 Rabbit IgG
B7-H3 (D9M2L) XP® Rabbit mAb #14058 20 µl W, IHC-Bond, IHC-P H 90 Rabbit IgG
4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb #34594 20 µl W, IP, IHC-P, IF-IC, F H 32, 40 Rabbit IgG
OX40 (E9U7O) XP® Rabbit mAb #61637 20 µl W, IP, IHC-Bond, IHC-P H, M, R 35-50 Rabbit IgG
GITR (D9I9D) Rabbit mAb (IHC Preferred) #68014 20 µl W, IHC-Bond, IHC-P H Mk 25 Rabbit IgG
CD40 Ligand (D5J9Y) Rabbit mAb #15094 20 µl W, IP, IHC-P H 25-30 (membrane bound); 17 (soluble) Rabbit IgG

†Species predicted to react based on 100% sequence homology.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-Bond=IHC-Leica® Bond™, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key: H=Human, Mk=Monkey, M=Mouse, R=Rat

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using B7-H3 (D9M2L) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines and tissues using CD40 Ligand (D5J9Y) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from OVKATE, DU 145, and MCF7 cells using VISTA ((D1L2G™) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from primary human CD4+ T cells and various cell lines using TIM-3 (D5D5R™) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). CD4+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).

Western Blotting

Western Blotting

Western blot analysis of extracts from HDLM-2, HuT 78, and Jurkat cells using LAG3 (D2G4O™) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from human CD4+ T cells, MOLT-4, and Jurkat cells using PD-1 (D4W2J) XP® Rabbit mAb (upper), and β-Actin (D6A8) Rabbit mAb #8457 (lower). CD4+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of human interleukin-2 (hIL-2) #8907 (6.7 ng/ml).


Western Blotting

Western Blotting

Western blot analysis of extracts from human CD8+ T cells, HuT 102, and Jurkat cells using GITR (D9I9D) Rabbit mAb (IHC Preferred) (upper), and β-Actin (D6A8) Rabbit mAb #8457 (lower). CD8+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of human interleukin-2 (hIL-2) #8907 (6.7 ng/ml).

IHC-Leica® Bond™

IHC-Leica® Bond™

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using GITR (D9I9D) Rabbit mAb (IHC Preferred) performed on the Leica® BOND™ Rx.

IHC-Leica® Bond™

IHC-Leica® Bond™

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using VISTA (D1L2G™) XP® Rabbit mAb performed on the Leica® BOND™ Rx.


IHC-Leica® Bond™

IHC-Leica® Bond™

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using B7-H3 (D9M2L) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

IHC-Leica® Bond™

IHC-Leica® Bond™

Immunohistochemical analysis of paraffin-embedded renal clear cell carcinoma using TIM-3 (D5D5R™) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

IHC-Leica® Bond™

IHC-Leica® Bond™

Immunohistochemical analysis of paraffin-embedded human breast ductal carcinoma using LAG3 (D2G4O™) XP® Rabbit mAb performed on the Leica® Bond™ Rx.


IHC-Leica® Bond™

IHC-Leica® Bond™

Immunohistochemical analysis of paraffin-embedded human infiltrating papillary carcinoma of the breast using PD-1 (D4W2J) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

Western Blotting

Western Blotting

Western blot analysis of extracts from primary human CD8+ T cells, treated with anti-CD3 and anti-CD28 in the presence of human interleukin-2, and various cell lines using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various endogenous human and mouse cells using OX40 (E9U7O) XP® Rabbit mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower). CD4+ T cells were purified from human blood or mouse spleens, and stimulated for 9 days using beads coated with human or mouse CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (20 ng/ml) or Mouse Interleukin-2 (mIL-2) #5201 (20 ng/ml).


Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human OX40 (Myc/DDK Human OX40; +) or mouse OX40 (Myc/DDK Mouse OX40; +), using OX40 (E9U7O) XP® Rabbit mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells (blue) and primary CD4+ T cells (green) using TIM-3 (D5D5R™) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. CD4+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded lung carcinoma using B7-H3 (D9M2L) XP® Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from COS-7 cells, mock transfected (-) or transfected with constructs expressing Myc-tagged full-length human PD-L1, PD-L2, B7-H3, or B7-H4 protein (as indicated), using B7-H3 (D9M2L) XP® Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of indicated amounts of recombinant Human CD40 Ligand (hCD40L) #3583 using CD40 Ligand (D5J9Y) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human kidney using VISTA (D1L2G™) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human tonsil using TIM-3 (D5D5R™) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with a construct expressing full-length human Myc/DDK-tagged TIM-3 (hTIM-3-Myc/DDK; +), using TIM-3 (D5D5R™) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HDLM-2 (left) and PC-3 (right) cell pellets on SignalSlide® PD-L1 IHC Controls #13747 using LAG3 (D2G4O™) XP® Rabbit mAb.


IP

IP

Immunoprecipitation of PD-1 protein from Molt-4 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is PD-1 (D4W2J) XP® Rabbit mAb. Western blot analysis was performed using PD-1

(D4W2J) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using PD-1 (D4W2J) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using GITR (D9I9D) Rabbit mAb (IHC Preferred).


IP

IP

Immunoprecipitation of 4-1BB/CD137/TNFRSF9 from HDLM-2 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb. Western blot analysis was performed using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.

IHC-Leica® Bond™

IHC-Leica® Bond™

Immunohistochemical analysis of paraffin-embedded human tonsil using OX40 (E9U7O) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded colon carcinoma using B7-H3 (D9M2L) XP® Rabbit mAb.


IP

IP

Immunoprecipitation of CD40 ligand from a HeLa cell lysate spiked with recombinant Human CD40 Ligand (hCD40L) #3583 (10 ng/ml) using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or CD40 Ligand (D5J9Y) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using CD40 Ligand (D5J9Y) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human tonsil using VISTA (D1L2G™) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using TIM-3 (D5D5R™) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colitis using LAG3 (D2G4O™) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using PD-1 (D4W2J) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human HuT 102 (left) and Jurkat (right) cell pellets using GITR (D9I9D) Rabbit mAb (IHC Preferred).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb.

IP

IP

Immunoprecipitation of OX40 protein from HuT 102 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is OX40 (E9U7O) XP® Rabbit mAb. Western blot analysis was performed using OX40 (E9U7O) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded ovarian carcinoma using B7-H3 (D9M2L) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).


IP

IP

Immunoprecipitation of CD40 ligand from Jurkat cells using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or CD40 Ligand (D5J9Y) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using CD40 Ligand (D5J9Y) Rabbit mAb. A conformation specific secondary antibody was used to avoid reactivity with IgG heavy and light chains.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's B-Cell lymphoma using VISTA (D1L2G™) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using TIM-3 (D5D5R™) XP® Rabbit mAb. Note staining of alveolar macrophages.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human tonsil using LAG3 (D2G4O™) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded 293 cell pellets, control (left) or PD-1 transfected (right), using PD-1 (D4W2J) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using GITR (D9I9D) Rabbit mAb (IHC Preferred).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human papillary carcinoma of the breast using OX40 (E9U7O) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded LNCaP (left) and NCI-H69 (right) cell pellets using B7-H3 (D9M2L) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human T-cell non-Hodgkin's lymphoma using CD40 Ligand (D5J9Y) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using VISTA (D1L2G™) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded cell pellets, primary CD4+ T cells (left) and HT-29 cells (right), using TIM-3 (D5D5R™) XP® Rabbit mAb. CD4+ T cells were purified from human blood and stimulated for 7 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HDLM-2 cell pellet (left, positive) or HT-29 cell pellet (right, negative) using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using OX40 (E9U7O) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Multiplex immunohistochemical analysis of paraffin-embedded human breast carcinoma usng PD-1 (D4W2J) XP® rabbit mAb (green), PD-L1 (E1L3N®) XP® rabbit mAb #13684 (red), LAG3 (D2G4O™) XP® rabbit mAb #15372 (magenta), TIM-3 (D5D5R™) XP® rabbit mAb #45208 (yellow), CD8α (C8/144B) mouse mAb #70306 (orange), and Pan-keratin (C11) mouse mAb #4545 (cyan).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded Jurkat (left) and Raji (right) cell pellets using CD40 Ligand (D5J9Y) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using VISTA (D1L2G™) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human thymus using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Multiplex immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using PD-1 (D4W2J) XP® rabbit mAb (green), B7-H3 (D9M2L) XP® rabbit mAb #14058 (red), B7-H4 (D1M8I) XP® rabbit mAb (cyan), LAG3 (D2G4O™) XP® rabbit mAb #15372 (magenta), TIM-3 (D5D5R™) XP® rabbit mAb #45208 (yellow), and VISTA (D1L2G™) XP® rabbit mAb #64953 (orange).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using OX40 (E9U7O) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded OVKATE (left) and MCF7 (right) cell pellets using VISTA (D1L2G™) XP® Rabbit mAb.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of human peripheral blood mononuclear cells (green), treated with anti-CD3 (10 μg/ml, 72h) and anti-CD28 (5 μg/ml, 72h), using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed lines) co-stained with a CD8 antibody. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Analysis was performed on CD8+ cells.

IHC-P (paraffin)

IHC-P (paraffin)

Multiplex immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using TIM-3 (D5D5R™) XP® rabbit mAb, PD-1 (EH33) mouse mAb #43248 (green), CD8α (C8/144B) mouse mAb #70306 (magenta), CD68 (D4B9C) XP® rabbit mAb #76437 (red), Pan-keratin (C11) mouse mAb #4545 (cyan),and LAG3 (D2G4O™) XP® rabbit mAb #15372 (orange).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using OX40 (E9U7O) XP® Rabbit mAb.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (left column) or CD3/CD28-treated (72 hr; right column), using PD-1 (D4W2J) XP® Rabbit mAb #86163 (top row) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (bottom row), and co-stained with a CD3 antibody.  Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells (blue) and HDLM-2 cells (green), using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HuT 78 cell pellet (left, positive) or Jurkat cell pellet (right, negative) using OX40 (E9U7O) XP® Rabbit mAb.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of human whole blood cells stained with VISTA (D1L2G™) XP® Rabbit mAb and co-stained with CD19-PE and CD14-APC. VISTA is present on CD14+ monocytes (right; upper right quadrant) and shows a significant increase in intensity when compared to its isotype control (middle; upper left quadrant). CD19+ B cells are negative for VISTA (left; upper left quadrant). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of primary human CD8+ T cells treated with anti-CD3 and anti-CD28 in the presence of human interleukin-2 (left, positive) and HeLa cells (right, negative) using 4-1BB/CD137/TNFRSF9 (D2Z4Y) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse spleen using OX40 (E9U7O) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded rat thymus using OX40 (E9U7O) XP® Rabbit mAb.

Description

The Human T Cell Co-inhibitory and Co-stimulatory Receptor IHC Antibody Sampler Kit provides an economical means of detecting expression of receptors that modulate T cell activity in formalin-fixed, paraffin-embedded tissue samples.

Specificity / Sensitivity

Each antibody included in the Human T Cell Co-inhibitory and Co-stimulatory Receptor IHC Antibody Sampler Kit recognizes endogenous levels of its target protein. CD40 Ligand (D5J9Y) Rabbit mAb recognizes endogenous levels of total membrane bound and soluble CD40 ligand protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala274 of human PD-1 protein, Gln264 of human OX40 protein, Val142 of human GITR protein, Ala94 of human B7-H3 protein, or near the carboxy terminus of human VISTA protein. Monoclonal antibodies are produced by immunizing animals with a recombinant protein specific to the extracellular domain of human TIM-3 protein, human 4-1BB/CD137/TNFRSF9 protein, human CD40 ligand protein, or the amino terminus of human LAG3 protein.

Background

PD-1 (PDCD1, CD279), TIM-3 (HAVCR2), LAG3 (CD223), VISTA (PD-H1), and B7-H3 (CD276) are immune cell co-inhibitory receptors (also known as immune checkpoints) that negatively regulate T cell function, and dampen the immune response to pathogens and cancer. In addition to activated T cells, PD-1 is expressed by activated B-cells and monocytes. TIM-3 is expressed by exhausted T cells in the settings of chronic infection and cancer. Tumor-infiltrating macrophages and dendritic cells also express TIM-3. LAG3 is primarily expressed by activated CD4+ T cells, CD8+ T cells, FoxP3+ T regulatory cells (Tregs) and natural killer (NK) cells. Although primarily expressed by myeloid cells, VISTA is also expressed by CD4+, CD8+, and Treg cells. Research examining the biological function of B7-H3 suggested that B7-H3 can be both a positive and negative regulator of T cell response. B7-H3 is expressed by antigen presenting cells, activated T cells, and a few normal tissues, including placenta and prostate. Expression of B7-H3 is seen in several cancer types, including prostate, breast, colon, lung, and gastric cancers, and in endothelial cells from tumor associated vasculature. Therapeutic blockade of these immune checkpoint receptors is a promising strategy for neoplastic intervention by enabling anti-tumor immune responses (1-3).

4-1BB (TNFRSF9, CD137), GITR (TNFRSF18), OX40 (TNFRSF4, CD134), and CD40 ligand (CD40L, CD154, TRAP, gp39) are immune cell co-stimulatory receptors that promote effector T cell survival and activation, and enable optimal immune responses to pathogens. 4-1BB is expressed in activated CD4+ and CD8+ T cells, natural killer cells and dendritic cells. GITR is expressed constitutively at high levels on Tregs, at low levels on naive and memory T cells, and is induced upon T cell activation. Studies show GITR can also be induced on NK cells, macrophages, and DCs. GITR ligation has been shown to induce CD8+ T cell activation, cytoxicity, and memory T cell survival, and conversely inhibit Treg suppressive function while promoting effector T cell resistance to Treg suppression. OX40 is primarily expressed on activated CD4+ and CD8+ T cells, while CD40L is primarily expressed on the surface of T cells, but has also been reported in blood platelets, mast cells, basophils, NK cells, and B cells. Research studies show that agonists of these co-stimulatory receptors augment anti-tumor immunity in several cancer types. Due to the combined effects on both Treg suppression and effector cell activation, GITR represents a unique opportunity for immunotherapeutic intervention in cancer. These pathways are an important area of interest in the study of cancer, vascular diseases, and inflammatory disorders (4-7).

  1. Schildberg, F.A. et al. (2016) Immunity 44, 955-72.
  2. Anderson, A.C. et al. (2016) Immunity 44, 989-1004.
  3. Callahan, M.K. et al. (2016) Immunity 44, 1069-78.
  4. Ward-Kavanagh, L.K. et al. (2016) Immunity 44, 1005-19.
  5. Ara, A. et al. (2018) Immunotargets Ther 7, 55-61.
  6. Knee, D.A. et al. (2016) Eur J Cancer 67, 1-10.
  7. Chester, C. et al. (2018) Blood 131, 49-57.

Application References

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