PA5-17520

antibody from Invitrogen Antibodies

Targeting: BIRC5 API4, EPR-1, survivin

Antibody data

Submit

Validation data

BIRC5 protein structure - PA5-17520 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: CRISPR-Cas9 mediated genome editing ofSurvivin (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR711998_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of Survivin was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) and HeLa Cas9 cells transduced with Survivin Lentiviral sgRNA (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 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 Anti-Survivin Polyclonal Antibody (Product # PA5-17520) using 1:1,000 dilution and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036 1:5,000 dilution).Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toSurvivin (Fig (b)).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Survivin in extracts from HT29 and Jurkat cells using Survivin polyclonal antibody (Product # PA5-17520).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Survivin Polyclonal Antibody (Product # PA5-17520) and a 18 kDa band corresponding to Survivin was observed across the cell lines and tissues tested except in heart tissue. Whole cell extracts and tissue lysates (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), A-431 (Lane 3), SK-BR-3 (Lane 4), SK-BR-3 treated with LY294002 (10 uM for 24 hrs) (Lane 5), Mouse Lung (Lane 6), Rat Lung (Lane 7), Mouse Heart (Lane 8) and Rat Heart (Lane 9) were electrophoresed using Novex™ 16% Tricine 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 the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25 µg/ml, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).