PA5-28691

antibody from Invitrogen Antibodies

Targeting: CSK

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-28691

Provider

Invitrogen Antibodies

Product name

CSK Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: 293T, A431, H1299, HeLaS3, HepG2, Molt-4, Raji.

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using CSK Polyclonal Antibody (Product # PA5-28691). Sample (30 µg whole cell lysate). A: 293T. B: H1299. C: MOLT4. 7.5% SDS PAGE. CSK Polyclonal Antibody (Product # PA5-28691) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of CSK was achieved by transfecting SH-SY5Y cells with CSK specific siRNAs (Silencer® select Product # s3612). Western blot analysis (Fig. a) was performed using whole cell extracts from the SH-SY5Y knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with CSK Polyclonal Antibody (Product # PA5-28691, 1:1500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP (Product # A27036, 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 CSK. An uncharacterized band (*) was observed at 100 kDa.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CSK Polyclonal Antibody (Product # PA5-28691) and a 51 kDa band corresponding to CSK was observed across all the cell lines tested. Whole cell extracts (30 µg lysate) of HCT 116 (Lane 1), SW480 (Lane 2), MCF7 (Lane 3) and MDA-MB-231 (Lane 4) 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 (0.75 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of c-Src in paraformaldehyde-fixed HeLa cells using a c-Src polyclonal antibody (Product # PA5-28691) at a 1:200 dilution.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded HT29 xenograft , using c-Src (Product # PA5-28691) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.