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MXD3 / MAD3 Recombinant Rabbit Monoclonal Antibody [JE48-14]

Usd: 385

Specification

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Catalog# ET7109-76

MXD3 / MAD3 Recombinant Rabbit Monoclonal Antibody [JE48-14]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

MXD3 / MAD3 Recombinant Rabbit Monoclonal Antibody [JE48-14]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human MXD3 aa 76-206 / 206.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 23 kDa

Positive Control

MG-63 cell lysate, SK-OV-3 cell lysate, Ramos cell lysate, HeLa cell lysate, Neuro-2a cell lysate, C6 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, human testis tissue.

Conjugation

unconjugated

Clone Number

JE48-14

RRID

AB_3070952

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

  • 1:2,000-1:5,000

  • IHC-P

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

MXD3 is a member of the MXD family of basic-helix-loop-helix transcription factors that form heterodimers with the cofactor MAX, and were originally proposed as functional antagonists of MYC proteins in the MYC/MAX/MAD transcriptional network. While some MXD proteins are MYC antagonists by acting as transcriptional repressors to promote cell differentiation, MXD3 is an atypical member that plays a role in proliferation rather than differentiation. MXD3 promotes granule neuron precursor (GNP) proliferation through a Sonic Hedgehog-mediated pathway, it is expressed in a mouse model of medulloblastoma, and its knock-down (KD) reduced proliferation of the human medulloblastoma cell line DAOY.

Background References

1. Barisone GA et al. Loss of MXD3 induces apoptosis of Reh human precursor B acute lymphoblastic leukemia cells. Blood Cells Mol Dis 54(4):329-335 (2015).

Subcellular Location

Nucleus.

UNIPROT

SwissProt: Q9BW11 Human

SwissProt: Q80US8 Mouse

SwissProt: Q62912 Rat

Synonyms

bHLHc13 antibody

Class C basic helix-loop-helix protein 13 antibody

FLJ35523 antibody

Likely ortholog of mouse Max dimerization protein 3 antibody

MAD3_HUMAN antibody

MAX dimerization protein 3 antibody

Max dimerizer 3 antibody

Max-associated protein 3 antibody

Max-interacting transcriptional repressor MAD3 antibody

MGC2383 antibody

Expand

Images

  • Western blot analysis of MXD3 / MAD3 on different lysates with Rabbit anti-MXD3 / MAD3 antibody (<a href="/products/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</a>) at 1/5,000 dilution.<br /><br />Lane 1: MG-63 cell lysate<br />Lane 2: SK-OV-3 cell lysate<br />Lane 3: Ramos cell lysate<br />Lane 4: HeLa cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 23 kDa<br />Observed band size: 27 kDa<br /><br />Exposure time: 8 seconds; ECL: K1801;<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/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</a>) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 MXD3 / MAD3 on different lysates with Rabbit anti-MXD3 / MAD3 antibody (ET7109-76) at 1/5,000 dilution.

    Lane 1: MG-63 cell lysate
    Lane 2: SK-OV-3 cell lysate
    Lane 3: Ramos cell lysate
    Lane 4: HeLa cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 23 kDa
    Observed band size: 27 kDa

    Exposure time: 8 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 (ET7109-76) at 1/5,000 dilution was used in primary antibody dilution (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.

  • Western blot analysis of MXD3 / MAD3 on different lysates with Rabbit anti-MXD3 / MAD3 antibody (<a href="/products/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</a>) at 1/2,000 dilution.<br /><br />Lane 1: Neuro-2a cell lysate<br />Lane 2: C6 cell lysate<br />Lane 3: Mouse testis tissue lysate<br />Lane 4: Rat testis tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 23 kDa<br />Observed band size: 27 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<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/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</a>) at 1/2,000 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 MXD3 / MAD3 on different lysates with Rabbit anti-MXD3 / MAD3 antibody (ET7109-76) at 1/2,000 dilution.

    Lane 1: Neuro-2a cell lysate
    Lane 2: C6 cell lysate
    Lane 3: Mouse testis tissue lysate
    Lane 4: Rat testis tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 23 kDa
    Observed band size: 27 kDa

    Exposure time: 20 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 (ET7109-76) at 1/2,000 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 testis tissue with Rabbit anti-MXD3 / MAD3 antibody (<a href="/products/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</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/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</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 testis tissue with Rabbit anti-MXD3 / MAD3 antibody (ET7109-76) 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 (ET7109-76) 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.

  • MXD3 / MAD3 was immunoprecipitated from 0.2 mg SK-OV-3 cell lysate with <a href="/products/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</a> at 1/2,000 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-OV-3 cell lysate (input)<br />Lane 2: <a href="/products/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</a> IP in SK-OV-3 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET7109-76" style="font-weight: bold;text-decoration: underline;">ET7109-76</a> in SK-OV-3 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute 50 seconds; ECL: K1801

    MXD3 / MAD3 was immunoprecipitated from 0.2 mg SK-OV-3 cell lysate with ET7109-76 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7109-76 at 1/2,000 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-OV-3 cell lysate (input)
    Lane 2: ET7109-76 IP in SK-OV-3 cell lysate
    Lane 3: Rabbit IgG instead of ET7109-76 in SK-OV-3 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute 50 seconds; ECL: K1801

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