PA5-17782

antibody from Invitrogen Antibodies

Targeting: RELA NFKB3, p65

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-17782

Provider

Invitrogen Antibodies

Product name

Phospho-NFkB p65 (Ser536) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Concentration

88 µg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Phospho-NFkB p65 (Ser536) Polyclonal Antibody (Product # PA5-17782) and a 65 kDa band corresponding to Transcription factor p65 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with TNF-alpha (20 ng/mL for 10 mins) and Calyculin A (100 nM for 10 min) (Lane 2), HeLa treated with LPS (1 µg/mL for 15 mins) and Calyculin A (100 nM for 10 min) (Lane 3), Jurkat (Lane 4), Jurkat treated with TNF-alpha (20 ng/mL for 10 mins) and Calyculin A (100 nM for 10 min) (Lane 5), THP-1 (Lane 6), THP-1 treated with LPS (1 µg/mL for 15 mins) and Calyculin A (100 nM for 10 min) (Lane 7), RAW 264.7 (Lane 8), RAW 264.7 treated with LPS (1 µg/mL for 15 mins) and Calyculin A (100 nM for 10 min) (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).