Human DNMT3B Antibody
R&D Systems, part of Bio-Techne | Catalog # AF7646
Key Product Details
Validated by
Knockout/Knockdown
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Immunocytochemistry, Knockout Validated, Simple Western, Western Blot
Cited:
Immunoprecipitation, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
Product Specifications
Immunogen
E. coli-derived recombinant human DNMT3B
Ser145-Pro355
Accession # Q9UBC3
Ser145-Pro355
Accession # Q9UBC3
Specificity
Detects human DNMT3B in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human DNMT3A is observed.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
Scientific Data Images for Human DNMT3B Antibody
Detection of Human DNMT3B by Western Blot.
Western blot shows lysates of K562 human chronic myelogenous leukemia cell line, BG01V human embryonic stem cells, and human thymus tissue. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human DNMT3B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7646) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for DNMT3B at approximately 100-110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
DNMT3B in BG01V Human Embryonic Stem Cells.
DNMT3B and SSEA-4 were detected in immersion fixed BG01V human embryonic stem cells. DNMT3B was detected using Sheep Anti-Human DNMT3B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7646) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Sheep IgG Secondary Antibody (green; Catalog # NL012). SSEA-4 was detected using Mouse Anti-Human/Mouse SSEA-4 Monoclonal Antibody (Catalog # MAB1435) and stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007). Specific staining of DNMT3B was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human DNMT3B by Simple WesternTM.
Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for DNMT3B at approximately 102 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human DNMT3B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7646) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.Applications for Human DNMT3B Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed BG01V human embryonic stem cells
Sample: Immersion fixed BG01V human embryonic stem cells
Knockout Validated
DNMT3B
is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in
DNMT3B knockout HeLa cell line.
Simple Western
10 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line
Sample: K562 human chronic myelogenous leukemia cell line
Western Blot
0.2 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line, BG01V human embryonic stem cells, and human thymus tissue
Sample: K562 human chronic myelogenous leukemia cell line, BG01V human embryonic stem cells, and human thymus tissue
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: DNMT3B
DNMT3B (DNA methyltransferase 3B; also DNA MTase HsaIIIB) is a 95-110 kDa member of the C5-methyltransferase family of enzymes. It is ubiquitously expressed in the embryo, and found associated with nuclear pericentric heterochromatin. Pericentric heterochromatin is chromatin composed of satellite repeats that lie adjacent to the centromere. DNA (cytosine-guanine dinucleotides) associated with this area undergos methylation, contributing to gene repression and proper chromosome segregation. Human DNMT3B is 853 amino acids (aa) in length. It contains a PWWP (Pro-Trp-Trp-Pro) domain that binds DNA (aa 225-283), a ADD (ATRX, DNMT3, DNMT3L) domain that contains a Zn-finger and binds to an unmethylated histone H3 tail (aa 423-555), and a class I AdoMet-MTase catalytic region that uses S-adenosyl-L-homocysteine as a substrate for methyl transfer (aa 577-847). There are at least five isoform variants. All show a deletion of aa 365-375. In addition, a second contains an alternative start site 12 aa upstream of the standard site, a third shows a deletion of aa 745-807, a fourth contains a Ser substitution for aa 744-853, while a fifth possesses a 45 aa substitution for aa 768-853. Over aa 1-355, human DNMT3B shares 73% aa sequence identity with mouse DNMT3B.
Long Name
DNA [Cytosine-5-]-Methyltransferase 3B
Alternate Names
ICF1, M.HsaIIIB
Gene Symbol
DNMT3B
UniProt
Additional DNMT3B Products
Product Documents for Human DNMT3B Antibody
Product Specific Notices for Human DNMT3B Antibody
For research use only
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