730044

antibody from Invitrogen Antibodies

Targeting: EP300 KAT3B, p300

Immunoprecipitation (Recommended by provider)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

730044

Provider

Invitrogen Antibodies

Product name

Anti-p300 Monoclonal Antibody, Biotin

Antibody type

Monoclonal

Antigen

Other

Reactivity

Human, Mouse

Host

Mouse

Conjugate

Biotin

Isotype

IgG

Vial size

100 µg

Concentration

0.5 mg/ml

Storage

Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C

References

Submitted references

Assessment of a method to characterize antibody selectivity and specificity for use in immunoprecipitation.

Marcon E, Jain H, Bhattacharya A, Guo H, Phanse S, Pu S, Byram G, Collins BC, Dowdell E, Fenner M, Guo X, Hutchinson A, Kennedy JJ, Krastins B, Larsen B, Lin ZY, Lopez MF, Loppnau P, Miersch S, Nguyen T, Olsen JB, Paduch M, Ravichandran M, Seitova A, Vadali G, Vogelsang MS, Whiteaker JR, Zhong G, Zhong N, Zhao L, Aebersold R, Arrowsmith CH, Emili A, Frappier L, Gingras AC, Gstaiger M, Paulovich AG, Koide S, Kossiakoff AA, Sidhu SS, Wodak SJ, Gräslund S, Greenblatt JF, Edwards AM
Nature methods 2015 Aug;12(8):725-31

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin immunoprecipitation analysis of p300 in mouse kidney was performed using biotinylated p300 antibody (Product # 730044). Kidney preparation: CD1 mice (Charles River Laboratories, Wilmington, MA; 6-10 weeks of age, male; 30-35 gms), received a tail vein injection of either LPS (2 mg/kg; 0111:B4; in 80 µl of PBS) or PBS. Two hours after injection, mice were anesthetized, the abdominal cavity was opened, the kidneys were extracted, and renal cortical samples were cut from the kidneys with a razor blade and rapidly frozen at -70oC. The mice died within 30 sec due to exsanguinations. Chromatin preparation and multiplex Matrix ChIP platform: The multiplex microplate Matrix ChIP method was previously described (see reference for Matrix ChIP protocol: (http://www.ncbi.nlm.nih.gov/pubmed/22098709). Briefly, kidney fragments (10-20 mg) were cross-linked with formaldehyde, and chromatin was sheared. ChIP assays were done using either protein A- or streptavidin-coated 96-well polypropylene microplates. p300-biotin antibody (Product # 730044) was used at 0.5 ug/well. For Pol II antibody (0.5ug/well) testing, wells were pre-incubated with 1ug/well of goat anti-mouse biotinylated antibody (Product # B-2763). 1ul of eluted DNA was used in 2 ul real-time PCR reactions (ABI7900HT) reactions containing primers shown to amplify exon 1 and exon 7 of the ICAM1 gene. PCR calibration curves were generated for each primer pair from a dilution series of total mouse or human genomic D

Conjugate

Biotin