711990

antibody from Invitrogen Antibodies

Targeting: CAT

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

711990

Provider

Invitrogen Antibodies

Product name

Catalase Recombinant Polyclonal Antibody (3HCLC)

Antibody type

Polyclonal

Antigen

Other

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

3HCLC

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Catalase was achieved by transfecting MCF7 cells with Catalase specific validated siRNA (Silencer® select Cat # s2443). Western blot analysis was performed using membrane 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 Polyclonal Antibody (Product # 711990, 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). 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 Hep G2 (Lane 1), K-562 (Lane 2) and tissue extracts (30 µg lysate) of Mouse Liver (Lane 3) and Rat Liver (Lane 4). The blots were probed with Anti-Catalase Recombinant Rabbit Polyclonal Antibody (Product # 711990, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 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 Hep G2 (Lane 1), K-562 (Lane 2) and tissue extracts (30 µg lysate) of Mouse Liver (Lane 3) and Rat Liver (Lane 4). The blots were probed with Anti-Catalase Recombinant Rabbit Polyclonal Antibody (Product # 711990, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 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).