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Carbonic anhydrase 2 Mouse Monoclonal Antibody [11A2]

Usd: 350

Specification

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Catalog# EM1801-14

Carbonic anhydrase 2 Mouse Monoclonal Antibody [11A2]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Carbonic anhydrase 2 Mouse Monoclonal Antibody [11A2]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within Human Carbonic anhydrase 2 aa 63-240 / 260.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 29 kDa

Positive Control

HL-60 cell lysate, HEK-293 cell lysate, A431 cell lysate, THP-1, human kidney tissue, rat liver tissue, human colon tissue, human stomach cancer tissue, mouse brain tissue.

Conjugation

unconjugated

Clone Number

11A2

RRID

AB_3068837

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG2a

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:100-1:2,000

Target

Function

Carbonic anhydrases (CAs) are members of a large family of zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide. CAs are involved in a variety of biological processes including respiration, calcification, acid-base balance and bone resorption, as well as the formation of aqueous humor, cerebrospinal fluid, saliva and gastric juice. They show extensive diversity in distribution and in their subcellular localization. The human CA2 gene, which maps to chromosome 8q21, encodes CA II, a cytoplasmic protein that has the highest turnover rate and widest tissue distribution of any known human CA isozyme. The human CA4 gene, which maps to chromosome 17q23, encodes CA IV, a membrane-anchored isozyme that is expressed on the luminal surfaces of pulmonary capillaries and proximal renal tubules. The human CA9, CA12 and CA14 genes, which map to chromosomes 9p13, 15q22 and 1q21, respectively, encode transmembrane proteins that have unique patterns of tissue-specific expression.

Background References

1. Tachibana H et al. Carbonic anhydrase 2 is a novel invasion-associated factor in urinary bladder cancers. Cancer Sci 108:331-337 (2017).

2. Gorrieri G et al. Goblet Cell Hyperplasia Requires High Bicarbonate Transport To Support Mucin Release. Sci Rep 6:36016 (2016).

Sequence Similarity

Belongs to the alpha-carbonic anhydrase family.

Subcellular Location

Cell membrane, Cytoplasm, Membrane.

UNIPROT

SwissProt: P00918 Human

SwissProt: P00920 Mouse

SwissProt: P27139 Rat

Synonyms

can antibody

CAN_ECOLI antibody

Carbonate dehydratase 2 antibody

Carbonic anhydrase 2 antibody

Images

  • Western blot analysis of Carbonic anhydrase 2 on different lysates with Mouse anti-Carbonic anhydrase 2 antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/1,000 dilution.<br /><br />Lane 1: HL-60 cell lysate<br />Lane 2: HEK-293 cell lysate<br />Lane 3: A431 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 29 kDa<br />Observed band size: 29 kDa<br /><br />Exposure time: 40 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Carbonic anhydrase 2 on different lysates with Mouse anti-Carbonic anhydrase 2 antibody (EM1801-14) at 1/1,000 dilution.

    Lane 1: HL-60 cell lysate
    Lane 2: HEK-293 cell lysate
    Lane 3: A431 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 29 kDa
    Observed band size: 29 kDa

    Exposure time: 40 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1801-14) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of THP-1 cells labeling Carbonic anhydrase 2 with Mouse anti-Carbonic anhydrase 2 antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/100 dilution.<br /><br />Cells were fixed in 80% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Carbonic anhydrase 2 antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of THP-1 cells labeling Carbonic anhydrase 2 with Mouse anti-Carbonic anhydrase 2 antibody (EM1801-14) at 1/100 dilution.

    Cells were fixed in 80% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Carbonic anhydrase 2 antibody (EM1801-14) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-Carbonic anhydrase 2 antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-Carbonic anhydrase 2 antibody (EM1801-14) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1801-14) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue using anti-Carbonic anhydrase 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat liver tissue using anti-Carbonic anhydrase 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (EM1801-14) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-Carbonic anhydrase 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-Carbonic anhydrase 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (EM1801-14) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue using anti-Carbonic anhydrase 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue using anti-Carbonic anhydrase 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (EM1801-14) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Carbonic anhydrase 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the antibody (<a href="/products/EM1801-14" style="font-weight: bold;text-decoration: underline;">EM1801-14</a>) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Carbonic anhydrase 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (EM1801-14) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

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