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GM2A Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# HA500560

GM2A Rabbit Polyclonal Antibody

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

GM2A Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Recombinant protein within human GM2A aa 1-193.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 21 kDa

Positive Control

SK-Br-3 cell lysate, HeLa cell lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, human kidney tissue, rat kidney tissue.

Conjugation

unconjugated

Reactivity Data

WB IHC-P IP
Human
Tested Tested
Tested Tested
Tested Tested
Mouse
Tested Tested
Not Recommended
Rat
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

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:200-1:1,000

  • IP

  • 1-2μg/sample

Target

Function

GM2 ganglioside activator also known as GM2A is a protein which in humans is encoded by the GM2A gene. The protein encoded by this gene is a small glycolipid transport protein which acts as a substrate specific co-factor for the lysosomal enzyme β-hexosaminidase A. β-hexosaminidase A, together with GM2 ganglioside activator, catalyzes the degradation of the ganglioside GM2, and other molecules containing terminal N-acetyl hexosamines. GM2A is a lipid transfer protein that stimulates the enzymatic processing of gangliosides, and also T-cell activation through lipid presentation. This protein binds molecules of ganglioside GM2, extracts them from membranes, and presents them to beta-hexosaminidase A for cleavage of N-acetyl-D-galactosamine and conversion to GM3. It was identified as a member of ML domain family of proteins involved in innate immunity and lipid metabolism in the SMART database.

Background References

1. Hsieh YC et al. Elevated ganglioside GM2 activator (GM2A) in human brain tissue reduces neurite integrity and spontaneous neuronal activity. Mol Neurodegener. 2022 Sep

2. Yuan J et al. Label-free quantitative proteomics reveals the Steap3-Gm2a axis inhibiting the phagosomal escape of Listeria monocytogenes. Microbes Infect. 2022 Nov-Dec

Subcellular Location

Lysosome.

UNIPROT

SwissProt: P17900 Human

SwissProt: Q60648 Mouse

Entrez Gene: 282838 Rat

Synonyms

Cerebroside sulfate activator protein antibody

ganglioside GM2 activator antibody

Ganglioside GM2 activator isoform short antibody

Ganglioside GM2 activator precursor antibody

GM2 activator antibody

GM2 AP antibody

GM2 ganglioside activator antibody

GM2 ganglioside activator protein antibody

GM2-AP antibody

GM2A antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of GM2A on different lysates with Rabbit anti-GM2A antibody (<a href="/products/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</a>) at 1/5,000 dilution.<br /><br />Lane 1: SK-Br-3 cell lysate (20 µg/Lane)<br />Lane 2: MDA-MB-231 cell lysate (low expression) (20 µg/Lane)<br />Lane 3: HeLa cell lysate (20 µg/Lane)<br />Lane 4: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 5: Mouse skeletal muscle tissue lysate (negative) (40 µg/Lane)<br />Lane 6: Rat kidney tissue lysate (40 µg/Lane)<br />Lane 7: Rat skeletal muscle tissue lysate (negative) (40 µg/Lane)<br /><br />Predicted band size: 21 kDa<br />Observed band size: 18 kDa<br /><br />Exposure time: 1 minute 30 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/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of GM2A on different lysates with Rabbit anti-GM2A antibody (HA500560) at 1/5,000 dilution.

    Lane 1: SK-Br-3 cell lysate (20 µg/Lane)
    Lane 2: MDA-MB-231 cell lysate (low expression) (20 µg/Lane)
    Lane 3: HeLa cell lysate (20 µg/Lane)
    Lane 4: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 5: Mouse skeletal muscle tissue lysate (negative) (40 µg/Lane)
    Lane 6: Rat kidney tissue lysate (40 µg/Lane)
    Lane 7: Rat skeletal muscle tissue lysate (negative) (40 µg/Lane)

    Predicted band size: 21 kDa
    Observed band size: 18 kDa

    Exposure time: 1 minute 30 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 (HA500560) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-GM2A antibody (<a href="/products/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</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/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</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 kidney tissue with Rabbit anti-GM2A antibody (HA500560) 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 (HA500560) 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 skeletal muscle tissue (negative) with Rabbit anti-GM2A antibody (<a href="/products/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</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/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</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 skeletal muscle tissue (negative) with Rabbit anti-GM2A antibody (HA500560) 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 (HA500560) 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 rat kidney tissue with Rabbit anti-GM2A antibody (<a href="/products/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</a>) at 1/400 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/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</a>) at 1/400 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 kidney tissue with Rabbit anti-GM2A antibody (HA500560) at 1/400 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 (HA500560) at 1/400 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 rat skeletal muscle tissue (negative) with Rabbit anti-GM2A antibody (<a href="/products/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</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/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</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 rat skeletal muscle tissue (negative) with Rabbit anti-GM2A antibody (HA500560) 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 (HA500560) 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.

  • GM2A was immunoprecipitated from 0.2 mg SK-Br-3 cell lysate with <a href="/products/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</a> at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: SK-Br-3 cell lysate (input)<br />Lane 2: <a href="/products/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</a> IP in SK-Br-3 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA500560" style="font-weight: bold;text-decoration: underline;">HA500560</a> in SK-Br-3 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 4 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    GM2A was immunoprecipitated from 0.2 mg SK-Br-3 cell lysate with HA500560 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA500560 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: SK-Br-3 cell lysate (input)
    Lane 2: HA500560 IP in SK-Br-3 cell lysate
    Lane 3: Rabbit IgG instead of HA500560 in SK-Br-3 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 4 seconds; ECL: K1801

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