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Carbonic anhydrase 9 Recombinant Rabbit Monoclonal Antibody [JJ088-9]

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Specification

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Catalog# ET1701-51

Carbonic anhydrase 9 Recombinant Rabbit Monoclonal Antibody [JJ088-9]

Application

  • WB

  • IHC-P

  • IF-Tissue

  • mIHC

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Carbonic anhydrase 9 Recombinant Rabbit Monoclonal Antibody [JJ088-9]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Carbonic anhydrase 9 aa 184-276 / 459.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Tissue, mIHC

Molecular Weight

Predicted band size: 50 kDa

Positive Control

Human lung tissue lysates, human liver tissue, human stomach tissue, human renal clear cell carcinoma tissue.

Conjugation

unconjugated

Clone Number

JJ088-9

RRID

AB_3070219

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*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:200-1:500

  • IF-Tissue

  • 1:100

  • mIHC

  • 1:100

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. CA IX is specifically expressed in clear-cell renal carcinomas, whereas CA XII is highly expressed in normal tissues, such as kidney, colon and pancreas. Human CA XIV is also expressed in normal tissues, such as brain, but differs from CA XII in its expression pattern.

Background References

1. Selfridge, A.C. et al. 2016. Hypercapnia Suppresses the HIF-dependent Adaptive Response to Hypoxia. The Journal of biological chemistry. 291: 11800-8.

2. Xue, G. et al. 2015. c-Myc-mediated repression of miR-15-16 in hypoxia is induced by increased HIF-2α and promotes tumor angiogenesis and metastasis by upregulating FGF2. Oncogene. 34: 1393-406.

Sequence Similarity

Belongs to the alpha-carbonic anhydrase family.

Tissue Specificity

Expressed primarily in carcinoma cells lines. Expression is restricted to very few normal tissues and the most abundant expression is found in the epithelial cells of gastric mucosa.

Post-translational Modification

Asn-346 bears high-mannose type glycan structures.

Subcellular Location

Nucleus, Cell membrane, Cell projection.

UNIPROT

SwissProt: Q16790 Human

Synonyms

CA-IX antibody

CA9 antibody

CAH9_HUMAN antibody

CAIX antibody

Carbonate dehydratase IX antibody

Carbonic anhydrase 9 antibody

Carbonic anhydrase IX antibody

Carbonic dehydratase antibody

G250 antibody

Membrane antigen MN antibody

Expand

Images

  • Fluorescence multiplex immunohistochemical analysis of human gastric cancer (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-LC3B (<a href="/products/ET1701-65" style="font-weight: bold;text-decoration: underline;">ET1701-65</a>, Green), anti-CD31 (<a href="/products/M1511-8" style="font-weight: bold;text-decoration: underline;">M1511-8</a>, Red) and anti-CA-IX (<a href="/products/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>, Yellow) on human gastric cancer. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKitCmTSA Kit <a href="/products/900802" style="font-weight: bold;text-decoration: underline;">900802</a>). The section was incubated in three rounds of staining: in the order of <a href="/products/ET1701-65" style="font-weight: bold;text-decoration: underline;">ET1701-65</a> (1/100 dilution), <a href="/products/M1511-8" style="font-weight: bold;text-decoration: underline;">M1511-8</a> (1/2,000 dilution) and <a href="/products/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a> (1/100 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

    Fluorescence multiplex immunohistochemical analysis of human gastric cancer (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-LC3B (ET1701-65, Green), anti-CD31 (M1511-8, Red) and anti-CA-IX (ET1701-51, Yellow) on human gastric cancer. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKitCmTSA Kit 900802). The section was incubated in three rounds of staining: in the order of ET1701-65 (1/100 dilution), M1511-8 (1/2,000 dilution) and ET1701-51 (1/100 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

  • Western blot analysis of Carbonic anhydrase 9 on human lung tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:40,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Carbonic anhydrase 9 on human lung tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-51, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Carbonic anhydrase 9 antibody (<a href="/products/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>) at 1/200 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 liver tissue with Rabbit anti-Carbonic anhydrase 9 antibody (ET1701-51) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1701-51) at 1/200 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 stomach tissue with Rabbit anti-Carbonic anhydrase 9 antibody (<a href="/products/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>) at 1/500 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 stomach tissue with Rabbit anti-Carbonic anhydrase 9 antibody (ET1701-51) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1701-51) at 1/500 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 renal clear cell carcinoma tissue with Rabbit anti-Carbonic anhydrase 9 antibody (<a href="/products/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>) at 1/200 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 renal clear cell carcinoma tissue with Rabbit anti-Carbonic anhydrase 9 antibody (ET1701-51) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1701-51) at 1/200 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.

  • Immunofluorescence analysis of paraffin-embedded human stomach tissue labeling Carbonic anhydrase 9 with Rabbit anti-Carbonic anhydrase 9 antibody (<a href="/products/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/ET1701-51" style="font-weight: bold;text-decoration: underline;">ET1701-51</a>, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of paraffin-embedded human stomach tissue labeling Carbonic anhydrase 9 with Rabbit anti-Carbonic anhydrase 9 antibody (ET1701-51) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1701-51, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

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Citation