Product Pathways - NF-kB Signaling
IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814
PhosphoSitePlus® protein, site, and accession data: IkB-alpha
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IHC-P IF-IC F | H M R Mk B Pg GP | Endogenous | 39 | Mouse IgG1 |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
B=Bovine
Pg=Pig
GP=Guinea Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) detects endogenous levels of total IκBα protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a GST-IκBα fusion protein corresponding the amino-terminus of human IκBα.
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-IκBα (Ser32/36) (5A5) Mouse mAb #9246 (upper) and IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (lower).
Western Blotting
Western blot analysis of extracts from HeLa, NIH/3T3 and PC12 cells, using IkBα (L35A5) Mouse mAb (Amino-terminal Antigen).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human leiomyoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human renal adenocarcinoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen).
Flow Cytometry
Flow cytometric analysis of HeLa cells, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (blue) compared to a nonspecifc negative control antibody (red).
IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left), or TNF-α-treated (#8902, 10 ng/ml for 15 min, right) using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Background
The NF-κB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IκB proteins (1-3). Activation occurs via phosphorylation of IκBα at Ser32 and Ser36 followed by proteasome-mediated degradation that results in the release and nuclear translocation of active NF-κB (3-7). IκBα phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals including inflammatory cytokines, growth factors, and chemokines. Kinases that phosphorylate IκB at these activating sites have been identified (8).
- Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6.
- Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70.
- Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
- Brown, K. et al. (1995) Science 267, 1485-8.
- Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
- Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
- Chen, Z.J. et al. (1996) Cell 84, 853-62.
- Karin, M. and Ben-Neriah, Y. (2000) Annu Rev Immunol 18, 621-63.
Application References
- Cui, W. et al. (2007) Proc Natl Acad Sci U S A 104, 14436-41. Applications: Western Blotting
- Shimada, M. et al. (2008) Development 135, 2001-11. Applications: Western Blotting
- Harrison, L.M. et al. (2008) Infect Immun , . Applications: Western Blotting
- Mabilleau, G. and Sabokbar, A. (2009) PLoS One 4, e4173. Applications: Western Blotting
- Kwong, C. et al. (2011) J Immunol 186, 1781-9. Applications: Western Blotting
- Barisic, S. et al. (2010) Biochem Pharmacol 80, 439-47. Applications: 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!
Companion Products
- 4812 IκBα (44D4) Rabbit mAb
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- 9247 IκBα (112B2) Mouse mAb (Carboxy-terminal Antigen)
- 2859 Phospho-IκBα (Ser32) (14D4) Rabbit mAb
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- 9248 IκBβ Antibody
- 4921 Phospho-IκBβ (Thr19/Ser23) Antibody (Human Specific)
- 9249 IκBε Antibody
- 4924 Phospho-IκBε (Ser18/22) Antibody
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- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 8902 Human Tumor Necrosis Factor-α (hTNF-α)
- 8125 SignalStain® Boost IHC Detection Reagent (HRP, Mouse)
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
