702732

antibody from Invitrogen Antibodies

Targeting: CAT

Antibody data

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Validation data

Product number: 702732 - Provider product page
Provider: Invitrogen Antibodies
Product name: Catalase Recombinant Rabbit Monoclonal Antibody (3H3L29)
Antibody type: Monoclonal
Antigen: Other
Description: This antibody predicted to react with Monkey, Bovine, Pig, Sheep
Antibody clone number: 3H3L29
Concentration: 0.5 mg/mL

CAT protein structure - 702732 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Catalase was achieved by transfecting MCF-7 cells with Catalase specific validated siRNA (Silencer® select Product # s2443). Western blot analysis (Fig a) was performed using Membrane enriched extracts from the Catalase knock down cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blots were probed with Anti-Catalase Recombinant Rabbit Monoclonal Antibody (Product # 702732, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Densitometric analysis of this Western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to Catalase.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on Membrane enriched extracts (30 µg lysate) of MCF-7 (Lane 1), HeLa (Lane 2), Jurkat (Lane 3), K-562 (Lane 4), A549 (Lane 5), HL-60 (Lane 6), HEK-293 (Lane 7) and tissue extracts of Mouse Liver (Lane 8). The blots were probed with Anti-Catalase Recombinant Rabbit Monoclonal Antibody (Product # 702732, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 59 kDa band corresponding to Catalase was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on Membrane enriched extracts (30 µg lysate) of MCF-7 (Lane 1), HeLa (Lane 2), Jurkat (Lane 3), K-562 (Lane 4), A549 (Lane 5), HL-60 (Lane 6), HEK-293 (Lane 7) and tissue extracts of Mouse Liver (Lane 8). The blots were probed with Anti-Catalase Recombinant Rabbit Monoclonal Antibody (Product # 702732, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 59 kDa band corresponding to Catalase was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).