Revision 5
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, ChIP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

50 Active form. 120 Precursor

Source/Isotype:

Rabbit IgG

UniProt ID:

#P19838

Entrez-Gene Id:

4790

Product Information

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:400 - 1:1600
Chromatin IP 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #78869.

Specificity / Sensitivity

NF-κB1 p105/p50 (D7H5M) Rabbit mAb recognizes endogenous levels of total NF-κB1 p105/p50 protein.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly415 of human NF-κB p105/p50 protein.

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).
Following IKK-mediated phosphorylation of p105 NF-κB at multiple sites (Ser921, 923, 927, and 932) on its carboxy-terminus, SCF/β-TrCP mediated processing produces the 50 kDa active form p50 (12,13).

  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.
  12. Heissmeyer, V. et al. (2001) Mol Cell Biol 21, 1024-35.
  13. Orian, A. et al. (2000) EMBO J 19, 2580-91.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 5
#12540

NF-κB1 p105/p50 (D7H5M) Rabbit mAb

Western Blotting Image 1: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
Western Blotting Image 2: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (10 ng/ml) for the indicated times, using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
No image available
Immunohistochemistry Image 1: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
Immunohistochemistry Image 2: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
Immunohistochemistry Image 3: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
Immunohistochemistry Image 4: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
Immunohistochemistry Image 5: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human thymus using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
Immunohistochemistry Image 6: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse spleen using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
Immunohistochemistry Image 7: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse thymus using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
Immunohistochemistry Image 8: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma (left), normal human spleen (middle), or 4T1 syngeneic mammary tumor (right) using NF-κB1 p105/p50 (D7H5M) Rabbit mAb (top) or NF-κB1 p105/p50 Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on NF-κB1 p105/p50 protein. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 9: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human appendix using NF-κB1 p105/p50 (D7H5M) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 10: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded MEF cell pellet, wild-type (left, positive) or NF-κB1 knockout (right, negative), using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.
Chromatin Immunoprecipitation Image 1: NF-κB1 p105/p50 (D7H5M) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30 ng/ml) for 1 hour and either NF-κB1 p105/p50 (D7H5M) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using SimpleChIP® Human IκBα Promoter Primers #5552, human IAP2 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.