MA5-16327

antibody from Invitrogen Antibodies

Targeting: CAST

Flow cytometry

Antibody data

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Validation data

Product number: MA5-16327 - Provider product page
Provider: Invitrogen Antibodies
Product name: Calpastatin Monoclonal Antibody (SP82)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is breast ductal carcinoma.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Antibody clone number: SP82
Vial size: 500 µL
Storage: 4° C, do not freeze

CAST protein structure - MA5-16327 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Calpastatin was achieved by transfecting HeLa cells with Calpastatin specific siRNAs (Silencer® select Product # s2398, s2399). Western blot analysis (Fig. a) was performed using whole cell extracts from Calpastatin knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Calpastatin Monoclonal Antibody (SP82) (Product # MA5-16327, 1:500 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 Calpastatin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Calpastatin Monoclonal Antibody (SP82) (Product # MA5-16327) and bands at 80-130 kDa corresponding to Calpastatin was observed in all the cell lines tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1) and HeLa (Lane 2) were electrophoresed using Novex® NuPAGE® 4-12% % Bis-Tris 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 the primary antibody (1:500 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)..