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MGEA5 Recombinant Rabbit Monoclonal Antibody [JG40-05]

Usd: 385

Specification

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Catalog# ET7109-01

MGEA5 Recombinant Rabbit Monoclonal Antibody [JG40-05]

Application

  • WB

  • IHC-P

  • IP

  • IF-Tissue

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

MGEA5 Recombinant Rabbit Monoclonal Antibody [JG40-05]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human MGEA5 aa 2-103 / 916.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP, IF-Tissue

Molecular Weight

Predicted band size: 103 kDa

Positive Control

PC-3M cell lysate, SH-SY5Y cell lysate, human tonsil tissue, human pancreas tissue, mouse testis tissue.

Conjugation

unconjugated

Clone Number

JG40-05

RRID

AB_3070891

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*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

  • 1:50-1:200

  • IP

  • 1-2μg/sample

  • IF-Tissue

  • 1:200

Target

Function

Cleaves GlcNAc but not GalNAc from O-glycosylated proteins. Can use p-nitrophenyl-beta-GlcNAc and 4-methylumbelliferone-GlcNAc as substrates but not p-nitrophenyl-beta-GalNAc or p-nitrophenyl-alpha-GlcNAc (in vitro). Does not bind acetyl-CoA and does not have histone acetyltransferase activity. Cleaves GlcNAc but not GalNAc from O-glycosylated proteins. Can use p-nitrophenyl-beta-GlcNAc as substrate but not p-nitrophenyl-beta-GalNAc or p-nitrophenyl-alpha-GlcNAc (in vitro), but has about six times lower specific activity than isoform 1.

Background References

1. Gao Y et al. Dynamic O-glycosylation of nuclear and cytosolic proteins: cloning and characterization of a neutral, cytosolic beta-N-acetylglucosaminidase from human brain. J Biol Chem 276:9838-9845 (2001).

2. Wells L et al. Dynamic O-glycosylation of nuclear and cytosolic proteins: further characterization of the nucleocytoplasmic beta-N-acetylglucosaminidase, O-GlcNAcase. J Biol Chem 277:1755-1761 (2002).

Sequence Similarity

Belongs to the glycosyl hydrolase 84 family.

Tissue Specificity

Ubiquitous. Shows highest expression in the brain, placenta and pancreas.

Post-translational Modification

Proteolytically cleaved by caspase-3 during apoptosis. The fragments interact with each other; cleavage does not decrease enzyme activity.

Subcellular Location

Cytoplasm. Nucleus.

UNIPROT

SwissProt: O60502 Human

SwissProt: Q9EQQ9 Mouse

Synonyms

Beta-hexosaminidase antibody

Beta-N-acetylglucosaminidase antibody

Beta-N-acetylhexosaminidase antibody

Bifunctional protein NCOAT antibody

HEXC antibody

Hyaluronidase in meningioma antibody

KIAA0679 antibody

MEA5 antibody

Meningioma expressed antigen 5 antibody

Meningioma-expressed antigen 5 antibody

Expand

Images

  • Western blot analysis of MGEA5 on different lysates with Rabbit anti-MGEA5 antibody (<a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: JAR cell lysate<br />Lane 3: HEK-293 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 103 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 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/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a>) at 1/1,000 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.

    Western blot analysis of MGEA5 on different lysates with Rabbit anti-MGEA5 antibody (ET7109-01) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: JAR cell lysate
    Lane 3: HEK-293 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 103 kDa
    Observed band size: 130 kDa

    Exposure time: 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 (ET7109-01) at 1/1,000 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of MGEA5 on different lysates with Rabbit anti-MGEA5 antibody (<a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-MGEA5 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 103 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 14 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/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a>) at 1/1,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> 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.

    ☑ Knockdown (KD)

    Western blot analysis of MGEA5 on different lysates with Rabbit anti-MGEA5 antibody (ET7109-01) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-MGEA5 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 103 kDa
    Observed band size: 130 kDa

    Exposure time: 14 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 (ET7109-01) at 1/1,000 dilution was used in K1803 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 colon tissue with Rabbit anti-MGEA5 antibody (<a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</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/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</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 colon tissue with Rabbit anti-MGEA5 antibody (ET7109-01) 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 (ET7109-01) 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 mouse testis tissue using anti-MGEA5 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 primary antibody (<a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a>, 1/50) for 30 minutes 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 mouse testis tissue using anti-MGEA5 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 primary antibody (ET7109-01, 1/50) for 30 minutes 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 mouse colon tissue with Rabbit anti-MGEA5 antibody (<a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a>) at 1/200 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/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</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 mouse colon tissue with Rabbit anti-MGEA5 antibody (ET7109-01) at 1/200 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 (ET7109-01) 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 rat colon tissue with Rabbit anti-MGEA5 antibody (<a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a>) at 1/200 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/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</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 rat colon tissue with Rabbit anti-MGEA5 antibody (ET7109-01) at 1/200 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 (ET7109-01) 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.

  • MGEA5 was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a> at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET7109-01" style="font-weight: bold;text-decoration: underline;">ET7109-01</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 30 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    MGEA5 was immunoprecipitated from 0.2 mg HeLa cell lysate with ET7109-01 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7109-01 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: ET7109-01 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of ET7109-01 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 30 seconds; ECL: K1801

  • Application: IF-Tissue<br /><br />Species: Mouse<br /><br />Site: colon<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/200

    Application: IF-Tissue

    Species: Mouse

    Site: colon

    Sample: Paraffin-embedded section

    Antibody concentration: 1/200

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