Product Pathways - NF-kappaB Signaling
PhosphoPlus® IκBα (Ser32/36) Antibody Kit #9240
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|---|---|
| Phospho-IκBα (Ser32/36) (5A5) Mouse mAb # 9246 | 100 microliters | W IP IHC-P E-P | H M R Mk Dg (Pg) (B) | 40 | Mouse |
| IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) # 4814 | 100 microliters | W IP IHC-P IF-IC F | H M R Mk Pg B | 39 | Mouse |
| NF-κB Control Cell Extracts # 9243 | 200 microliters | ||||
| Anti-mouse IgG, HRP-linked Antibody # 7076 | 50 microliters | Horse | |||
| Anti-biotin, HRP-linked Antibody # 7075 | 100 microliters | Goat | |||
| 20X LumiGLO® Reagent and 20X Peroxide # 7003 | 5 milliliters each | ||||
| Biotinylated Protein Ladder Detection Pack # 7727 | 100 microliters |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
E-P=ELISA (Peptide)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Pg=Pig
B=Bovine
Dg=Dog
Specificity / Sensitivity
IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 detects endogenous levels of total IκBα protein. Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 detects endogenous levels of IκBα only when phosphorylated at Ser32/36. Neither antibody cross-reacts with other IκB family members at physiological levels.
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) #4814 (lower).
Western Blotting
Western blot analysis of extracts from HeLa, NIH/3T3 and PC12 cells, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814.
Western Blotting
Western blot analysis of extracts from NIH/3T3 cells, untreated orTNF-α-treated (#2169, 20 ng/ml) for 5 minutes, using Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 (upper) or IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 (lower).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human leiomyoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human renal adenocarcinoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast tumor, using Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246.
IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left), or TNF-α-treated (right) using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
Source / Purification
IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 is produced by immunizing mice with a GST-IκBα fusion protein corresponding the amino-terminus of human IκBα. Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 is produced by immunizing mice with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser32/36 of human IκBα. Total cell extracts from HeLa cells prepared without treatment serve as a negative control. Supplied in SDS Sample Buffer. Total cell extracts from HeLa cells prepared with TNF-α treatment (#2169, 20 ng/ml for 5 minutes) serve as a positive control. Supplied in SDS Sample Buffer.
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
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
- 9246 Phospho-IκBα (Ser32/36) (5A5) Mouse mAb
- 2859 Phospho-IκBα (Ser32) (14D4) Rabbit mAb
- 4812 IκBα (44D4) Rabbit mAb
- 4814 IκBα (L35A5) Mouse mAb (Amino-terminal Antigen)
- 9242 IκBα Antibody
- 9247 IκBα (112B2) Mouse mAb (Carboxy-terminal Antigen)
- 2682 IKKα Antibody
- 2684 IKKβ Antibody
- 2678 IKKβ (L570) Antibody (IP Preferred)
- 2697 Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb
- 3034 NF-κB p65 Antibody
- 4764 NF-κB p65 (C22B4) Rabbit mAb
- 3033 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb
- 3037 Phospho-NF-κB p65 (Ser276) Antibody
- 3039 Phospho-NF-κB p65 (Ser468) Antibody
- 9243 NF-κB Control Cell Extracts
- 2169 Tumor Necrosis Factor-α
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
