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PSMA Mouse Monoclonal Antibody [A10A2]

Usd: 350

Specification

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Catalog# HA601169

PSMA Mouse Monoclonal Antibody [A10A2]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

PSMA Mouse Monoclonal Antibody [A10A2]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within human PSMA aa 20-750 / 750.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 84 kDa

Positive Control

LNCaP cell lysate, 22RV1 cell lysate, human prostate tissue, human prostate carcinoma tissue, LNCaP.

Conjugation

unconjugated

Clone Number

A10A2

RRID

AB_3071888

Reactivity Data

WB IHC-P IF-Cell
Human
Tested Tested
Tested Tested
Tested Tested

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

Glutamate carboxypeptidase II (GCPII), also known as N-acetyl-L-aspartyl-L-glutamate peptidase I (NAALADase I), NAAG peptidase, or prostate-specific membrane antigen (PSMA) is an enzyme that in humans is encoded by the FOLH1 (folate hydrolase 1) gene. Human GCPII contains 750 amino acids and weighs approximately 84 kDa. GCPII is a zinc metalloenzyme that resides in membranes. Most of the enzyme resides in the extracellular space. GCPII is a class II membrane glycoprotein. It catalyzes the hydrolysis of N-acetylaspartylglutamate (NAAG) to glutamate and N-acetylaspartate (NAA) according to the reaction scheme to the right. Neuroscientists primarily use the term NAALADase in their studies, while those studying folate metabolism use folate hydrolase, and those studying prostate cancer or oncology, PSMA. All refer to the same protein glutamate carboxypeptidase II.

Background References

1. Wang F et al. Advances in PSMA-targeted therapy for prostate cancer. Prostate Cancer Prostatic Dis. 2022 Mar

2. Seifert R et al. Prostate Cancer Theranostics: PSMA Targeted Therapy. PET Clin. 2021 Jul

Subcellular Location

Cell membrane, Cytoplasm.

UNIPROT

SwissProt: Q04609 Human

Synonyms

Cell growth inhibiting protein 27 antibody

Cell growth-inhibiting gene 27 protein antibody

FGCP antibody

Folate hydrolase (prostate-specific membrane antigen) 1 antibody

Folate hydrolase 1 antibody

Folate hydrolase antibody

Folate hydrolase prostate specific membrane antigen 1 antibody

FOLH 1 antibody

FOLH antibody

Folh1 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of PSMA on different lysates with Mouse anti-PSMA antibody (<a href="/products/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/1,000 dilution.<br /><br />Lane 1: LNCaP cell lysate<br />Lane 2: 22RV1 cell lysate<br />Lane 3: PC-3M cell lysate (negative)<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 84 kDa<br />Observed band size: 100/200 kDa<br /><br />Exposure time: 43 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/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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.

    ☑ Relative expression (RE)

    Western blot analysis of PSMA on different lysates with Mouse anti-PSMA antibody (HA601169) at 1/1,000 dilution.

    Lane 1: LNCaP cell lysate
    Lane 2: 22RV1 cell lysate
    Lane 3: PC-3M cell lysate (negative)

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 84 kDa
    Observed band size: 100/200 kDa

    Exposure time: 43 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 (HA601169) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of PSMA on different lysates with Mouse anti-PSMA antibody (<a href="/products/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/5,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-PSMA KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 84 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 180 seconds; ECL: K1801;<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/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/5,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> 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.

    ☑ Knockdown (KD)

    Western blot analysis of PSMA on different lysates with Mouse anti-PSMA antibody (HA601169) at 1/5,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-PSMA KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 84 kDa
    Observed band size: 110 kDa

    Exposure time: 180 seconds; ECL: K1801;

    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 (HA601169) at 1/5,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of LNCaP (positive) and PC-3M (negative) cells labeling PSMA with Mouse anti-PSMA antibody (<a href="/products/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/100 dilution.<br /><br />Cells were fixed in 100% 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-PSMA antibody (<a href="/products/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</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.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of LNCaP (positive) and PC-3M (negative) cells labeling PSMA with Mouse anti-PSMA antibody (HA601169) at 1/100 dilution.

    Cells were fixed in 100% 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-PSMA antibody (HA601169) 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 prostate tissue with Mouse anti-PSMA antibody (<a href="/products/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/1,000 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/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/1,000 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 prostate tissue with Mouse anti-PSMA antibody (HA601169) at 1/1,000 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 (HA601169) at 1/1,000 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 prostate carcinoma tissue with Mouse anti-PSMA antibody (<a href="/products/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/1,000 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/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/1,000 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 prostate carcinoma tissue with Mouse anti-PSMA antibody (HA601169) at 1/1,000 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 (HA601169) at 1/1,000 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunohistochemical analysis of paraffin-embedded human colon tissue (negative control) with Mouse anti-PSMA antibody (<a href="/products/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/1,000 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/HA601169" style="font-weight: bold;text-decoration: underline;">HA601169</a>) at 1/1,000 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.

    ☑ Relative expression (RE)

    Immunohistochemical analysis of paraffin-embedded human colon tissue (negative control) with Mouse anti-PSMA antibody (HA601169) at 1/1,000 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 (HA601169) at 1/1,000 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.

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