Analysis Note

ControlJurkat whole cell lysate, HeLa nuclear extract.

Included Positive Antigen Control:
Catalog # 12-309, HeLa nuclear extract. Add an equal volume of Laemmli reducing sample buffer to 5-10 µL of extract and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Research Sub CategoryTranscription Factors

Immunoprecipitation:
4 µg of a previous lot immunoprecipitated NFκB p52 and its p100 precursor from 500 µg of HeLa nuclear extract cell lysate.

Gel Shift Assay:
A previous lot of this antibody used 2 µg per gel shift assay (Bours, V., 1993).

Research CategoryEpigenetics & Nuclear Function

Anti-NFκB p52 Antibody is a Mouse Monoclonal Antibody for detection of NFκB p52 also known as DNA-binding factor KBF2, Lymphocyte translocation chromosome 10, Oncogene Lyt-10 & has been validated in EMSA, IP & WB.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

The NFκB transcription factor was originally identified as a protein complex consisting of a DNA binding subunit and an associated protein. The DNA binding subunit is functionally related to c-Rel p75 and Rel B p68. The p50 subunit was initially believed to be a functionally unique protein derived from the amino terminus of a precursor designated p105. A cDNA has been isolated that encodes an alternative DNA binding subunit of NFκB. It is synthesized as a protein that is expressed in a variety of cell types and, like p105, undergoes cleavage to generate its NFκB subunit, in this case a protein designated p52 (previously referred to as p49). In contrast to p50 derived from p105, p52 acts in synergy with p65 to stimulate the HIV enhancer in transiently transfected Jurkat cells.

Immunogen

GST-fusion protein corresponding to amino acids 1-444 of human NFκB p52 subunit (Schmidt, R., 1991;Bours, V., 1992).

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein G Chromatography

Purified mouse monoclonal IgG2a in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide.

Format: Purified

Quality

Routinely evaluated by immunoblotting in HeLa nuclear extract or in human Raji cell, but not in mouse 3T3/A31 cell and rat PC 12 cell lysates

Western Blot Analysis:
0.5-2 µg/mL of this lot detected NFκB p52 and its p100 precursor in HeLa nuclear extract. A previous lot detected NFκB p52 and its p100 precursor in human Raji cell, but not in mouse 3T3/A31 cell and rat PC 12 cell lysates. Highly recommended for western blotting.

Specificity

Does not cross-react with mouse or rat; other species cross-reactivity is unknown.

This antibody recognizes the p52 subunit of NFκB and its p100 precursor.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

52 kDa; precursor at 100 kDa