MA5-24108

antibody from Invitrogen Antibodies

Targeting: NAMPT PBEF, PBEF1

Western blot (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-24108

Provider

Invitrogen Antibodies

Product name

NAMPT Monoclonal Antibody (362616)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

In direct ELISAs and Western blots, 100% cross-reactivity with recombinant human PBEF is observed.

Antibody clone number

362616

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-NAMPT Monoclonal Antibody (362616)(Product # MA5-24108) and a 55 kDa band corresponding to NAMPT was observed across cell lines tested. Increased expression of NAMPT was observed in HUVEC upon IL1-beta and TNF-alpha treatment. Whole cell extracts (30 µg lysate) of HUVEC (Lane 1), HUVEC treated with IL1-beta (10 ng/mL for 18hr) (Lane 2), HUVEC treated with TNF-alpha (10 ng/mL for 18hr) (Lane 3) and U-87 MG (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 µg/mL) and detected by chemiluminescence with F(ab2-Rabbit anti-Rat IgG (H+L Secondary Antibody, HRP (Product # PA1-29927,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of NAMPT in 3T3‚L1 mouse embryonic fibroblast adipose‚like cell line. Samples were incubated in NAMPT monoclonal antibody (Product # MA5-24108) or isotype control antibody followed by Allophycocyanin-conjugated Anti-Rat IgG F(ab)2 Secondary Antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.