MA5-11940

antibody from Invitrogen Antibodies

Targeting: CHEK2 bA444G7, CDS1, CHK2, HuCds1, PP1425, RAD53

Immunoprecipitation

Antibody data

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Validation data

Product number: MA5-11940 - Provider product page
Provider: Invitrogen Antibodies
Product name: CHK2 Monoclonal Antibody (2CHK01 (DCS-293))
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-11940 targets Chk2 in immunofluorescence, immunoprecipiation, and Western blot applications and shows reactivity with Human samples. The antibody does not react with Chk1. The MA5-11940 immunogen is recombinant Chk2 protein.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 2CHK01 (DCS-293)
Vial size: 500 µL
Concentration: 0.2 mg/mL
Storage: 4° C

CHEK2 protein structure - MA5-11940 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Chk2 using Chk2 Monoclonal Antibody (Product # MA5-11940) on HeLa Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Serine/threonine-protein kinase Chk2 was achieved by transfecting HeLa with Serine/threonine-protein kinase Chk2 specific siRNAs (Silencer® select Product # S22121, S22119). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Serine/threonine-protein kinase Chk2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with CHK2 Monoclonal Antibody (2CHK01 (DCS-293)) (Product # MA5-11940, 1 µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 Serine/threonine-protein kinase Chk2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-CHK2 Monoclonal Antibody (2CHK01 (DCS-293)) (Product # MA5-11940) and a 60 kDa band corresponding to Serine/threonine-protein kinase Chk2 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HEK-293 (Lane 2), HT-29 (Lane 3), SH-SY5Y (Lane 4) and HCT 116 (Lane 5) 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 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of CHK2 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1020054_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of CHK2 was performed by loading 50µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2), and HeLa CHK2 KO (Lane 3) 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 Anti-CHK2 Monoclonal Antibody (2CHK01 (DCS-293)) (Product # MA5-11940, 1:100 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:5000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to CHK2. Uncharacterized bands were observed in HeLa Cas9 and KO samples at ~70kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEL 92.1.7 (Lane 1), and HEK-293 (Lane 2). The blots were probed with Anti-Chk2 Mouse Monoclonal Antibody (Product # MA5-11940, 1-3 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 62-6520, 1:4000 dilution). A 60 kDa band corresponding to Chk2 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).