MA5-49145

antibody from Invitrogen Antibodies

Targeting: CA2 CA-II, CAII, Car2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-49145

Provider

Invitrogen Antibodies

Product name

Carbonic Anhydrase II Monoclonal Antibody (10E11)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

Adding 0.2 mL of distilled water will yield a concentration of 500 µg/mL. Human CA2 shares 81.1% and 80.7% amino acid (aa) sequence identity with mouse and rat CA2, respectively. Positive Control - WB: rat heart tissue, rat kidney tissue, rat spleen tissue, rat liver tissue, mouse heart tissue, mouse kidney tissue, mouse spleen tissue, mouse liver tissue, human placenta tissue, human 293T whole cell. IHC: human gastric cancer tissue, human colon cancer tissue, mouse intestine tissue, rat intestine tissue. ICC/IF: HEPG2 cell. Flow: 293T cell.|Store at -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freeze-thaw cycles.

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

10E11

Vial size

100 µg

Concentration

500 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of CA2 using CA2 antibody (Product # MA5-49145). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 µg of sample under reducing conditions. Lane 1: rat heart tissue lysates; Lane 2: rat kidney tissue lysates; Lane 3: rat spleen tissue lysates; Lane 4: rat liver tissue lysates; Lane 5: mouse heart tissue lysates; Lane 6: mouse kidney tissue lysates; Lane 7: mouse spleen tissue lysates; Lane 8: mouse liver tissue lysates; Lane 9: human placenta tissue lysates; Lane 10: human 293T whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-CA2 antigen affinity purified monoclonal antibody (Product # MA5-49145) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10,000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CA2 at approximately 28KD. The expected band size for CA2 is at 28KD.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of Carbonic Anhydrase II in HEPG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. Samples were then incubated in Carbonic Anhydrase II Monoclonal antibody (Product # MA5-49145) using a dilution of 5 μg/mL. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Carbonic Anhydrase II in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with Carbonic Anhydrase II Monoclonal antibody (Product # MA5-49145) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Carbonic Anhydrase II in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with Carbonic Anhydrase II Monoclonal antibody (Product # MA5-49145) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Carbonic Anhydrase II in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with Carbonic Anhydrase II Monoclonal antibody (Product # MA5-49145) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Carbonic Anhydrase II in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with Carbonic Anhydrase II Monoclonal antibody (Product # MA5-49145) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Carbonic Anhydrase II in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with Carbonic Anhydrase II Monoclonal antibody (Product # MA5-49145) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of Carbonic Anhydrase II in 293T cells using Carbonic Anhydrase II Monoclonal Antibody (10E11) (Product # MA5-49145), shown in overlay histogram (blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum, and incubated with the primary antibody (1 μg/1x10^6 cells) for 30 min at 20°C. DyLight 488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.