ZRB2328

Anti-MBD3 Antibody, clone 1K19 ZooMAb^® Rabbit Monoclonal

recombinant, expressed in HEK 293 cells

• Synonym(s): Methyl-CpG-binding domain protein 3, Methyl-CpG-binding protein MBD3
• UNSPSC Code: 12352203
• NACRES: NA.41

Properties

• biological source: rabbit
• Quality Level: 200
• recombinant: expressed in HEK 293 cells
• conjugate: unconjugated
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 1K19, recombinant monoclonal
• description: 1K19 Clone
• product line: ZooMAb^® learn more
• form: lyophilized
• mol wt: calculated mol wt 32.84 kDa; observed mol wt ~32 kDa
• purified by: using Protein A
• species reactivity: human, mouse, rat
• species reactivity (predicted by homology): monkey
• packaging: antibody small pack of 25 μL
• greener alternative product characteristics: Waste Prevention; Designing Safer Chemicals; Design for Energy Efficiency; Learn more about the Principles of Green Chemistry.
• enhanced validation: recombinant expression; Learn more about Antibody Enhanced Validation
• technique(s): affinity binding assay: suitable; immunocytochemistry: suitable; immunohistochemistry: suitable; western blot: suitable
• isotype: IgG
• epitope sequence: N-terminal half
• Protein ID accession no.: NP_001268382
• UniProt accession no.: O95983
• greener alternative category: Aligned
• shipped in: ambient
• storage temp.: 2-8°C
• Gene Information: human ... MBD3(53615)

Description

General description

We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry. This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.

ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

Specificity

Clone 1K19 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects Methyl-CpG-binding protein MBD3. It targets an epitope within 18 amino acids from the N-terminal half.

Immunogen

KLH-conjugated linear peptide corresponding to 18 amino acids from the N-terminal half of human Methyl-CpG-binding protein MBD3.

Application

Quality Control Testing

Evaluated by Western Blotting in K562 cell lysate.

Western Blotting Analysis: A 1:10,000 dilution of this antibody detected MBD3 in K562 cell lysate.

Tested Applications

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected MBD3 in lysates from Mouse brain tissue and C6 cells and 1:10,000 dilution in HeLa cell lysate.

Affinity Binding Assay: A representative lot of this antibody bound MBD3 peptide with a KD of 3.9 x 10-8 in an affinity binding assay.

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected MBD3 in Human kidney tissue sections.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected MBD3 in HeLa cells.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Target description

Methyl-CpG-binding domain protein 3 (UniProt: O95983; also known as Methyl-CpG-binding protein MBD3, MBD3) is encoded by the MBD3 gene (Gene ID: 53615) in human. MBD3 is transcriptional repressor that plays a role in gene silencing. It can form heterodimer with MBD2 via an interaction between the coiled-coil domains of the two proteins. It is localized in discrete foci in the nucleus and is detected on chromatin, at promoter regions of active genes. MBD3 is a component of the nuRD/Mi2  complex that consists of Rpd3-like histone deacetylase, RbAp46, MTA2, CHD4/Mi2beta, and MBD3. This complex possesses DNA-dependent ATPase, chromatin remodeling activity, and histone deacetylase activity. The MBD3 displays high degree of homology with MBD2 and its MBD domain is localized in amino acids 1-72. Apart from their MBD and coiled coil domain, MBD2 contains an additional N-terminal glycine-arginine rich domain and a transcriptional repressor domain, whereas MBD3 has a C-terminal poly-glutamate region. In spite of having a MBD domain, MBD3 does not bind to mCpG specifically. It has been reported that histidine 30 and phenylalanine 34 are responsible for the inability of MBD3 to bind to mCpG. Mutant forms containing H30K/F34Y are shown to bind to mCpG in vitro. The MBD2-MBD3 heterodimer recognizes hemi-methylated DNA concurrent with DNA replication and recruits histone deacetylase complexes and DNA-methyltransferase 1 (DNMT1) to establish and/or maintain the transcriptionally repressed chromatin. MBD3 also plays a key role in lineage commitment as MBD3 depleted embryonic stem cells fail to differentiate and aberrantly self-renew independent of leukemia inhibitory factor (LIF). This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Menafra, R., and Stunnenberg, HG. (2014). Front. Genet. 5; 428; Saito, M., and Ishikawa, F. (2002). J. Biol. Chem. 277(38):35434-35439; Tatematsu, KI., et al. (2000). Genes Cells. 5(8); 677-688).

Physical form

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

Reconstitution

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety Information

• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable