MA5-14989

antibody from Invitrogen Antibodies

Targeting: CBX5 HP1, HP1-ALPHA, HP1Hs-alpha

Antibody data

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Validation data

CBX5 protein structure - MA5-14989 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Chromobox protein homolog 5 was achieved by transfecting HeLa with Chromobox protein homolog 5 specific siRNAs (Silencer® select Product # s23884, s23883). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the Chromobox protein homolog 5 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with HP1 alpha Monoclonal Antibody (F.747.6) (Product # MA5-14989, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant 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 Chromobox protein homolog 5.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-HP1 alpha Monoclonal Antibody (F.747.6) (Product # MA5-14989) and a 25 kDa band corresponding to Chromobox protein homolog 5 was observed across tested cell lines. Nuclear enriched extracts (40 µg lysate) of MOLT-4 (Lane 1), HeLa (Lane 2), SH-SY5Y (Lane 3), PC-3 (Lane 4), Hep G2 (Lane 5), NIH/3T3 (Lane 6) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). 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, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).