PA5-18358

antibody from Invitrogen Antibodies

Targeting: NPM1 B23, NPM

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Antibody Data

Product number

PA5-18358

Provider

Invitrogen Antibodies

Product name

NPM1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with bovine and canine based on sequence homology.

Reactivity

Human, Mouse

Host

Goat

Isotype

IgG

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot staining of HeLa cell lysate using Product # PA5-18358 at a concentration of 0.1 µg/mL, the primary antibody incubation was 1 hour and the detection method was chemiluminescence.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of NPM1 was achieved by transfecting HeLa with NPM1 specific siRNAs (Silencer® select Product # s9677, s9676). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from NPM1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with NPM1 Polyclonal Antibody (Product # PA5-18358, 1ug/mL) and Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to NPM1.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-NPM1 Polyclonal Antibody (Product # PA5-18358) and a 32 kDa band corresponding to NPM1 was observed across all the cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), COS-7 (Lane 2), Hep G2 (Lane 3), HEK-293 (Lane 4), MCF7 (Lane 5), and A-431 (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (1ug/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L) Superclonal™ Secondary Antibody, HRP (Product # A27014, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).