Catalog # HK6520

Histone H2B Phospho-Regulation Antibody Kit

Antibody Sampler Kit

DATASHEET  




$395

The nucleosome is a protein complex consisting of four core histones (H2A, H2B, H3, and H4). Two molecules of each histone forms an octamer that makes up the nucleosome. DNA wraps around repeating nucleosome units to generate chromatin structures. The structure of chromatin determines the accessiblity to transcription factors. Post-translational modification of the amino-terminal tail of histones in nucleosomes alters chromatin structure to promote or inhibit transcription. Complex alterations in acetylation, methylation, ubiquination, and/or phosphorylation determine the chromatin structural changes that occur during specific phases of the cell cycle or in response to cell stimuli. One mode of regulating histone H2B activity is through phosphorylation in the amino terminal region. Important sites of phosphorylation include Ser-14, Ser-32, and Ser-36. AMPK phosphorylates Ser-36 on histone H2B during cell stress leading to increased transcription and cell survival, while ectopic expression of an unphosphorylatable histone H2B during cell stress reduces transcription of AMPK-dependent genes and lowers cell survival.

References
Lau, A.T. et al. (2011) J Biol Chem. 286(30):26628.
Ajiro, K. et al. (2010) Cell Death Differ. 17(6):984.
Bungard, D. et al. (2010) Science. 329(5996):1201.
Workman, J.L. & Kingston, R.E. (1998) Annu Rev Biochem. 67:545.

Western blot analysis of Jurkat cells treated with calyculin A (100 nM) for 30 min. (lanes 1, 3, 5). The blots were treated with lambda phosphatase (lanes 2, 4, 6), then probed with anti-Histone H2B (C-terminus) (lanes 1 & 2), anti-Histone H2B (Ser-36) (lanes 3 & 4), and anti-Histone H2B (a.a. 33-47) (lanes 5 & 6).

Immunocytochemical labeling of Histone H2B in methanol and acetone fixed rat A7r5 cells. The cells were labeled with rabbit polyclonal Histone H2B (C-terminus) antibody (HP4291), then the antibody was detected using appropriate secondary antibody conjugated to DyLight® 594.



Rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.



*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

HP4331 Bertoldo, M. et al. (2016) Mol Cell Endocrin. 7207(16):30001 WB: mouse Sertoli cells
HP4331 Lee, J.H. et al. (2015) Nucleic acids res 43(9): 4505. WB: MCF10A
HP4291 Seo, Y. et al. (2013) PLoS ONE 8(8):e75005. WB: MDCK, A549, and HEK293
HP4291 Soe, K.C. et al. (2013) BBA 1829(11):1184. WB: Sf9 cells
HP4331 Soe, K.C. et al. (2013) BBA 1829(11):1184. WB: Sf9 cells
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
Histone H2B (C-terminus) Rabbit pAb50 μlWB, E, ICCHu, Rt, Ms, F15
Histone H2B (a.a. 33-47) Rabbit pAb50 μlWB, EHu, Rt, Ms15
Histone H2B (Ser-36), phospho-specific Rabbit pAb 50 μlWB, EHu, Rt, Ms15
Anti-Rabbit Ig Light Chain Specific:HRP Mouse mAb100 μlWB, E, ICC, IHCRb
KIT SUMMARY

The histone H2B phospho-regulation kit can be used to examine phosphorylation of Ser-36 compared to the total expression of Histone H2B. The kit includes two different rabbit polyconal antibodies to detect total Histone H2B, and secondary reagent for rabbit polyclonal antibody detection.