MA5-15207

antibody from Invitrogen Antibodies

Targeting: BIRC5 API4, EPR-1, survivin

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15207

Provider

Invitrogen Antibodies

Product name

Survivin Monoclonal Antibody (H.987.7)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody. This antibody is not cross-reactive with other types of apoptotic inhibitor proteins.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

H.987.7

Vial size

100 µL

Storage

-20°C

Western blot

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 Monoclonal Antibody (H.987.7) (Product # MA5-15207) using 1:1,000 dilution and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177 1:5,000 dilution).Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). 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 HeLa cells 48 hours following mock transfection, transfection with nonspecific (control) siRNA or transfection with Survivin siRNA, using Survivin monoclonal antibody (Product # MA5-15207) and a p42 MAP Kinase (Erk2) polyclonal antibody. The Survivin antibody confirms silencing of Survivin expression, and the p42 MAPK antibody is used to control for protein loading and siRNA specificity.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Survivin Monoclonal Antibody (H.987.7) (Product # MA5-15207) and a 17 kDa band corresponding to Survivin was observed across cell lines tested and was lost upon LY294002 treatment in SK-BR-3. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), U-2 OS (Lane 2), A549 (Lane 3), K-562 (Lane 4), A-431 (Lane 5), SK-BR-3 (Lane 6) and SK-BR-3 treated with LY294002 (10uM for 24 hours) (Lane 7) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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-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).