Anti–MBD1 polyclonal antibody

Catalog #

25303

$415 *

Size: 50 µg

Qty

Bulk/Custom

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Description

Polyclonal antibody raised in rabbit against human MBD1 (Methyl–CpG–binding domain protein 1), using a KLH–conjugated synthetic peptide containing a sequence from the N–terminal part of the protein.

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Synonyms

CXXC3, PCM1, RFT

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Product Data Gallery

Product Info

Storage and Usage

Citations

Clonality

polyclonal

Host Species/Expression System

Rabbit

Purification

Affinity purified

Format

Aqueous buffer solution

Formulation

PBS containing 0.05% azide and 0.05% ProClin 300

Concentration

50 µg/72 µl

Immunogen

synthetic peptide

Background

MBD1 (UniProt/Swiss-Prot entry Q9UIS9) is a transcriptional repressor that specifically binds to methylated CpG dinucleotides in promoter sequences. MBD1 acts by recruiting a variety of histone deacetylases (HDAC’s) and chromatin remodelling factors. MBD1-dependent transcriptional repression is mediated by ATF7IP through the recruitment of factors such as the histone methyltransferase SETDB1. MBD1 probably forms a complex with SETDB1 and ATF7IP which couples DNA methylation to H3K9 trimethylation and represses transcription.

Storage/Stability

Store at –80°C for up to 2 years. Centrifuge after first thaw to maximize product recovery. Aliquot to avoid repeated freeze/thaw cycles. Aliquots may be stored at –20°C for at least one month.

Assay Conditions

ChIP results obtained with the antibody directed against MBD1
ChIP assays were performed using human osteosarcoma (U2OS) cells, the antibody against MBD1 (Cat. # 25303) and optimized PCR primer sets. Sheared chromatin from 1x10⁶ cells and 1.5 µg of antibody were used per ChIP experiment. Beads only were used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the MLH1 gene (used as a positive control) and CDC6 gene (used as a negative control). Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against human MBD1 (Cat. # 25303), crude serum and flow-through. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the purified antibody was estimated to be 1:20,000.

Western blot analysis using the antibody directed against MBD1
Nuclear extracts of HeLa cells (40 µg) were analyzed by Western blot using the antibody against MBD1 (Cat. # 25303) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right, the marker (in kDa) is shown on the left.

Applications

ChIP (1.5 µg)
ELISA (1:1000)
WB (1:500)

Shipping Temperature

-20°C or-80°C

Species Reactivity

Human

Notes

The optimal antibody amount per IP should be determined by the end–user. We recommend testing 1–5 µg per IP

Warnings

Avoid freeze/thaw cycles

Scientific Category

Antibodies

Regulatory Status

Research Use Only

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Anti–MBD1 polyclonal antibody

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