New England Biolabs Canada
 

Product Pathways - NF-kB Signaling

NF-κB p65 (C22B4) Rabbit mAb #4764

Item# Description List Price Web Price Qty
4764S NF-KappaB p65 (C22B4) Rabbit mAb - 100 µl $372.00
$334.80
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4764T NF-κB p65 (C22B4) Rabbit mAb - 20 µl $159.00
$143.10
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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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1061S NF-κB p65 Blocking Peptide - 100 µg $458.00
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12629S Acetyl-NF-κB p65 (Lys310) (D2S3J) Rabbit mAb - 100 µl $433.00
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12630P SignalFire™ Plus ECL Reagent - 20 ml $116.00
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12757S SignalFire™ Elite ECL Reagent - 100 ml $558.00
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13188S Methyl-NF-κB p65 (Lys310) Antibody - 100 µl $433.00
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2859L Phospho-IKappaBAlpha (Ser32) (14D4) Rabbit mAb - 300 µl $1,025.00
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2859S Phospho-IKappaBAlpha (Ser32) (14D4) Rabbit mAb - 100 µl $433.00
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2859T Phospho-IκBα (Ser32) (14D4) Rabbit mAb - 20 µl $174.00
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3031L Phospho-NF-KappaB p65 (Ser536) Antibody - 300 µl $1,025.00
$922.50
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3031S Phospho-NF-κB p65 (Ser536) Antibody - 100 µl $433.00
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3033L Phospho-NF-KappaB p65 (Ser536) (93H1) Rabbit mAb - 300 µl $1,025.00
$922.50
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3033S Phospho-NF-KappaB p65 (Ser536) (93H1) Rabbit mAb - 100 µl $433.00
$389.70
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3033T Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb - 20 µl $174.00
$156.60
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3035S NF-κB1 p105/p50 Antibody - 100 µl $372.00
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3039S Phospho-NF-κB p65 (Ser468) Antibody - 100 µl $433.00
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3045S Acetyl-NF-κB p65 (Lys310) Antibody - 100 µl $433.00
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4812S IκBα (44D4) Rabbit mAb - 100 µl $372.00
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4814S IKappaBAlpha (L35A5) Mouse mAb (Amino-terminal Antigen) - 100 µl $372.00
$334.80
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4814T IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) - 20 µl $159.00
$143.10
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4882S NF-KappaB2 p100/p52 Antibody - 100 µl $372.00
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4882T NF-κB2 p100/p52 Antibody - 20 µl $159.00
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4954S RelB Antibody - 100 µl $372.00
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5425S Normal Goat Serum - 10 ml $92.00
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6883P3 SignalFire™ ECL Reagent - 50 ml $43.00
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6883S SignalFire™ ECL Reagent - 500 ml $370.00
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7074P2 Anti-rabbit IgG, HRP-linked Antibody - 100 µl $109.00
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7074S Anti-rabbit IgG, HRP-linked Antibody - 1 ml $203.00
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7074V Anti-rabbit IgG, HRP-linked Antibody - 5 ml $646.00
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7727L Biotinylated Protein Ladder Detection Pack - 3250 µl $326.00
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7727S Biotinylated Protein Ladder Detection Pack - 650 µl $92.00
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8112L SignalStain® Antibody Diluent - 100 ml $117.00
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8112S SignalStain® Antibody Diluent - 25 ml $57.00
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8114P SignalStain® Boost IHC Detection Reagent (HRP, Rabbit) - 1 ml $56.00
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8114S SignalStain® Boost IHC Detection Reagent (HRP, Rabbit) - 15 ml $525.00
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8242S NF-KappaB p65 (D14E12) XP® Rabbit mAb - 100 µl $419.00
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8242T NF-κB p65 (D14E12) XP® Rabbit mAb - 20 µl $174.00
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9243S NF-κB Control Cell Extracts - 200 µl $169.00
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9936T NF-κB Pathway Sampler Kit - 1 Kit $694.00
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9997S Tris Buffered Saline with Tween® 20 (TBST-10X) - 1000 ml $90.00
$81.00
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VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W Human, Mouse, Rat, Monkey, Bovine Endogenous 65 Rabbit IgG
IF-IC
F

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry

Homology

Species predicted to react based on 100% sequence homology: Dog.

Protocols

Specificity / Sensitivity

NF-κB p65 (C22B4) Rabbit mAb detects endogenous levels of total NF-κB p65 protein. This antibody may also cross-react with the p50 subunit.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human NF-κB p65.

Western Blotting

Western Blotting

Western blot analysis of extracts from THP-1 cells, differentiated with TPA (#9905, 80 nM for 24h) and treated with 1 μg/ml LPS for the indicated times, using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033 (upper) and NF-κB p65 (C22B4) Rabbit mAb (lower).

Western Blotting

Western Blotting

Western blot analysis of HeLa cell extracts, untreated (-) or NF-κB p65 knock-out (+), using NF-κB p65 (C22B4) Rabbit mAb #4764 (upper) or β-actin (13E5) Rabbit mAb #4970 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa (human), PC12 (rat) and Neuro2A (mouse) cell lines using NF-κB p65 (C22B4) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence® NF-κB p65 siRNA II (+), or SignalSilence® NF-κB p65 siRNA I #6261 (+), using NF-κB p65 (C22B4) Rabbit mAb #4764 and α-Tubulin (11H10) Rabbit mAb #2125. NF-κB p65 (C22B4) Rabbit mAb confirms silencing of NF-κB p65 and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of NF-κB p65 siRNA.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or TNF-α-treated (#8902, 20ng/ml for 20 min, right), using NF-κB p65 (C22B4) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of OVCAR8 cells using NF-κB p65 (C22B4) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).


Background

Transcription factors of the nuclear factor κB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11).

  1. Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.
  2. Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.
  3. Haskill, S. et al. (1991) Cell 65, 1281-9.
  4. Thompson, J.E. et al. (1995) Cell 80, 573-82.
  5. Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  7. Scherer, D.C. et al. (1995) Proc Natl Acad Sci USA 92, 11259-63.
  8. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  9. Senftleben, U. et al. (2001) Science 293, 1495-9.
  10. Coope, H.J. et al. (2002) EMBO J 21, 5375-85.
  11. Xiao, G. et al. (2001) Mol Cell 7, 401-9.

Application References

  • Hasler, U. et al. (2008) J Biol Chem 283, 28095-105. Applications: Western Blotting.
  • Hideshima, T. et al. (2009) Blood 113, 5228-36. Applications: IHC-P (paraffin), Western Blotting.
  • Xie, S. et al. (2010) J Immunol 184, 2289-96. Applications: Western Blotting.
  • Ikner, A. and Ashkenazi, A. (2011) J Biol Chem 286, 21546-54. Applications: Western Blotting.
  • Stairs, D.B. et al. (2011) Cancer Cell 19, 470-83. Applications: IC-IF, Western Blotting.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

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