PA1-16831

NUMB antibody from Invitrogen Antibodies

C14orf41

Western blot

Supportive data in Antibodypedia

Immunocytochemistry

Recommended by provider

Provider product page for PA1-16831

Antibody Data

Product number

PA1-16831

Provider

Invitrogen Antibodies

Product name

Anti-NUMB Polyclonal Antibody

Provider product page

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

The target sequence has 90% sequence homology with rat.

Reactivity

Human, Mouse, Chicken/Avian

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

3.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Detection of Numb isoforms 1 and 2 in A431 whole cell lysate (20 µg) using 0.5 µg/mL (Product # PA1-16831). ECL detection: 30 seconds.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of NUMB was achieved by transfecting U-87 MG cells with NUMB specific siRNAs (Silencer® select Product # s16466, s16465). Western blot analysis (Fig. a) was performed using whole cell extracts from the NUMB knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed using NUMB Polyclonal Antibody (Product # PA1-16831) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 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 NUMB isoforms.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-NUMB Polyclonal Antibody (Product # PA1-16831) on whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), PC-3 (Lane 2), DU 145 (Lane 3), A549 (Lane 4), SH-SY5Y (Lane 5), HeLa (Lane 6) and Mouse lung (Lane 7) and numb major isoform 65, 72 kDa was observed across cell lines except A549. 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 (1:2000 dilution) and detected by Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).