MA5-16413

antibody from Invitrogen Antibodies

Targeting: CA9 CAIX, MN

Flow cytometry

Antibody data

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Validation data

Product number: MA5-16413 - Provider product page
Provider: Invitrogen Antibodies
Product name: Carbonic Anhydrase IX Monoclonal Antibody (SP106)
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 10 min at room temperature. A suggested positive control is stomach tissue.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Antibody clone number: SP106
Vial size: 500 µL
Concentration: 0.11 mg/mL
Storage: 4° C, do not freeze

CA9 protein structure - MA5-16413 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of CA9 was achieved by transfecting U-87 MG with CA9 specific siRNAs (Silencer® select Product # s2269, s2270). Western blot analysis (Fig. a) was performed using whole cell extracts from the CA9 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Carbonic Anhydrase IX Monoclonal Antibody (SP106) (Product # MA5-16413, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25µg/ml, 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 CA9.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Carbonic Anhydrase IX Monoclonal Antibody (Product # MA5-16413) and ~ 50-55 kDa bands corresponding to CA9 was observed across cell lines along with an uncharacterized band (*) at ~80 kDa in the cell lines tested. Whole cell extracts (60 µg lysate) of U-87 MG (Lane 1) and HeLa (Lane 2) were electrophoresed using 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).