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MBD1 Recombinant Rabbit Monoclonal Antibody [PSH01-79] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750753

MBD1 Recombinant Rabbit Monoclonal Antibody [PSH01-79] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

MBD1 Recombinant Rabbit Monoclonal Antibody [PSH01-79] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse MBD1 aa 1-200 / 636.

Species Reactivity

Mouse, Rat

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 70 kDa

Positive Control

NIH/3T3 cell lysate, RAW264.7 cell lysate, C2C12 cell lysate, mouse kidney tissue lysate, mouse brain tissue lysate, mouse lung tissue lysate, mouse spleen tissue lysate, rat brain tissue lysate, NIH/3T3.

Conjugation

unconjugated

Clone Number

PSH01-79

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

MBD1 is a member of a family of nuclear proteins related by the presence of a methyl-CpG binding domain (MBD). These proteins are capable of binding specifically to methylated DNA, and some members can also repress transcription from methylated gene promoters. This protein contains multiple domains: MBD at the N-terminus that functions both in binding to methylated DNA and in protein interactions; several CXXC-type zinc finger domains that mediate binding to non-methylated CpG dinucleotides; transcriptional repression domain (TRD) at the C-terminus that is involved in transcription repression and in protein interactions. Numerous alternatively spliced transcript variants encoding different isoforms have been noted for this gene.

Background References

1. Wensheng L, Bo Z, Qiangsheng H, et al. MBD1 promotes the malignant behavior of gallbladder cancer cells and induces chemotherapeutic resistance to gemcitabine. Cancer Cell Int. 2019 Sep 9; 19:232.

2. Lyst MJ, Nan X, Stancheva I. Retraction: Regulation of MBD1-mediated transcriptional repression by SUMO and PIAS proteins. EMBO J. 2019 Oct 15;38(20): e103220.

Subcellular Location

Nucleus, Nucleus matrix, Nucleus speckle, Chromosome

UNIPROT

SwissProt: Q9Z2E2 Mouse

SwissProt: Q66HB8 Rat

Synonyms

CXXC 3 antibody

CXXC-type zinc finger protein 3 antibody

CXXC3 antibody

MBD 1 antibody

Mbd1 antibody

MBD1_HUMAN antibody

MECP1 COMPLEX antibody

Methyl CpG binding domain protein 1 antibody

Methyl CpG binding domain protein 1 isoform PCM1 antibody

Methyl CpG binding protein MBD1 antibody

Expand

Images

  • Western blot analysis of MBD1 on different lysates with Rabbit anti-MBD1 antibody (<a href="/products/HA750753" style="font-weight: bold;text-decoration: underline;">HA750753</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate<br />Lane 2: RAW264.7 cell lysate<br />Lane 3: C2C12 cell lysate<br />Lane 4: Mouse kidney tissue lysate<br />Lane 5: Mouse brain tissue lysate<br />Lane 6: Mouse lung tissue lysate<br />Lane 7: Mouse spleen tissue lysate<br />Lane 8: Rat brain tissue lysate<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 70 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 5 minutes;<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/HA750753" style="font-weight: bold;text-decoration: underline;">HA750753</a>) at 1/1,000 dilution was used in 5% NFDM/TBST 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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of MBD1 on different lysates with Rabbit anti-MBD1 antibody (HA750753) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate
    Lane 2: RAW264.7 cell lysate
    Lane 3: C2C12 cell lysate
    Lane 4: Mouse kidney tissue lysate
    Lane 5: Mouse brain tissue lysate
    Lane 6: Mouse lung tissue lysate
    Lane 7: Mouse spleen tissue lysate
    Lane 8: Rat brain tissue lysate

    Lysates/proteins at 30 µg/Lane.

    Predicted band size: 70 kDa
    Observed band size: 70 kDa

    Exposure time: 5 minutes;

    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 (HA750753) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling MBD1 with Rabbit anti-MBD1 antibody (<a href="/products/HA750753" style="font-weight: bold;text-decoration: underline;">HA750753</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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 Rabbit anti-MBD1 antibody (<a href="/products/HA750753" style="font-weight: bold;text-decoration: underline;">HA750753</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. 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. 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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NIH/3T3 cells labeling MBD1 with Rabbit anti-MBD1 antibody (HA750753) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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 Rabbit anti-MBD1 antibody (HA750753) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of NIH/3T3 cells labeling MBD1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA750753" style="font-weight: bold;text-decoration: underline;">HA750753</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of NIH/3T3 cells labeling MBD1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA750753, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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