MA5-15282

antibody from Invitrogen Antibodies

Targeting: PTK2 FADK, FAK, FAK1, PPP1R71

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15282

Provider

Invitrogen Antibodies

Product name

FAK Monoclonal Antibody (4A9D6)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA5-15282 targets FAK in WB applications and shows reactivity with Human samples.

Antibody clone number

4A9D6

Concentration

Conc. Not Determined

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of FAK using a FAK monoclonal antibody (Product # MA5-15282) against a truncated FAK recombinant protein.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of FAK using a FAK monoclonal antibody (Product # MA5-15282) against a truncated FAK recombinant protein.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

CRISPR-Cas9 mediated genome editing ofFAK (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR627416_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of FAK was performed by loading 30 µg of A549 Cas9 (Lane 1) and A549 Cas9 cells transduced with FAK Lentiviral sgRNA (Lane 2) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ Novex™ 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 FAK Monoclonal Antibody (4A9D6) (Product # MA5-15282, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10,000 dilution). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076) using the iBright™ FL 1500 (Product # A44115). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toFAK (Fig (b)). An uncharacterized band was observed at ~35 kDa in both the samples.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-FAK Monoclonal Antibody (4A9D6) (Product # MA5-15282) and a120kDa band corresponding to Focal adhesion kinase 1 was observed across cell lines tested except U-937 which is reported to be negative for FAK expression. Additional uncharacterized bands (*) were observed across the cell lines tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), HCT 116 (Lane 2), A549 (Lane 3), A-431 (Lane 4), SH-SY5Y (Lane 5), K-562 (Lane 6) and U-937 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) 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-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)