PA5-77880

antibody from Invitrogen Antibodies

Targeting: BLM BS, RECQ2, RECQL3

Antibody data

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Validation data

BLM protein structure - PA5-77880 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of BLM in A) 293T and B) A431 cells using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with BLM polyclonal antibody (Product # PA5-77880) using a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Blooms Syndrome Polyclonal Antibody (Product # PA5-77880). Whole cell extract (30 µg) was separated by 5% SDS-PAGE, and the membrane was blotted with Blooms Syndrome Polyclonal Antibody (Product # PA5-77880) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of Blooms Syndrome was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR815029_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Blooms Syndrome was performed by loading 30 µg of HeLa Cas9 (Lane 1) andHeLa Blooms Syndrome KO (Lane 2) modified whole cell extracts. The samples were electrophoresed using NuPAGE™ 3 to 8%, Tris-Acetate, 1.0 mm, Mini Protein Gel (Product # EC6695BOX). 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-Blooms Syndrome Polyclonal Antibody (Product # PA5-77880, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:5000 dilution) using the iBright™ FL 1500 (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Blooms Syndrome. Uncharacterized band was observed in all the samples at ~37 kDa.