Analysis Note

ControlTNFα treated HeLa cells, PMA, A431 cells or calcium ionophore-treated Jurkat cells
Included Positive Antigen Control: Catalog # 12-301, non-stimulated A431 lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate 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 p65 from 500 µg of a human A431 carcinoma cell lysate.

Research CategoryEpigenetics & Nuclear Function

Anti-NFκB p65 Antibody, CT is a Rabbit Polyclonal Antibody for detection of NFκB p65 also known as Nuclear factor NF-kappa-B p65 subunit & 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 transcription factor NFκB (Nuclear Factor κB) is involved in the expression and regulation of a number of important cellular and physiological processes such as growth, development, apoptosis, immune and inflammatory response, and activation of various viral promoters including human immunodeficiency virus long terminal repeats. NFκB represents a group of structurally related and evolutionarily conserved proteins related to the proto-oncogene c-Rel with five members in mammals that include Rel (cRel), RelA (p65), RelB, NFκB1 (p50 and its precursor p105), and NFκB2 (p52 and its precursor p100). NFκB/Rel proteins exist as homo- or heterodimers to form transcriptionally competent or repressive complexes. Although most NFκB dimers are activators of transcription, the p50/50 and p52/52 homodimers can repress the transcription of their target genes. The p50/p65 heterodimer of NFκB is the most abundant in cells.

Immunogen

Epitope: C-terminus

KLH-conjugated, synthetic peptide corresponding to the 13 C-terminal amino acids (C-DMDFSALLSQISS) of the p65 subunit of human NFκB containing an N-terminal cysteine for conjugation. The immunizing sequence is identical in mouse.

Legal Information

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

Linkage

Replaces: 04-1008; 04-235

Other Notes

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

Physical form

Purified rabbit polyclonal IgG in 0.014 M phosphate buffer, pH 7.6, 0.175 M NaCl, 0.07% sodium azide and 30% glycerol. Liquid at -20°C.

Protein A chromatography

Format: Purified

Quality

Routinely evaluated by Western Blot on RIPA lysates from A431 cells.

Western Blot Analysis:
0.5-2 µg/mL of this lot detected NFkB p65 in RIPA lysates from A431 cells.

Specificity

NFκB p65, Mr 65 kDa

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. Note: Variability in freezer temperatures below -20°C may cause glycerolcontaining solutions to become frozen during storage.

Target description

65 kDa