DNMT3B antibody (DNA (Cytosine-5-)-Methyltransferase 3 beta)

Details for Product anti-DNMT3B Antibody No. ABIN251114, Supplier: Log in to see
Antigen
  • cb633
  • dnmt3b
  • DNMT3B
  • LOC100218113
  • ICF
  • ICF1
  • M.HsaIIIB
  • cb91
  • dnmt3bl
  • dnmt7
  • sb:cb91
  • wu:fb16h07
  • MmuIIIB
  • DNA (cytosine-5-)-methyltransferase 4
  • DNA (cytosine-5-)-methyltransferase 3 beta
  • DNA methyltransferase 3B
  • dnmt4
  • DNMT3B
  • LOC100218113
  • dnmt3b
  • Dnmt3b
Reactivity
Human, Mouse (Murine)
123
63
27
4
2
2
2
2
2
2
2
1
1
1
Host
Rabbit
87
38
2
2
2
Clonality
Polyclonal
Conjugate
This DNMT3B antibody is un-conjugated
4
4
4
3
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Application
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Immunohistochemistry (IHC), Western Blotting (WB)
111
46
39
38
34
32
24
16
15
13
10
3
3
3
2
2
2
2
1
Options
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Immunogen Amino acids 4-101 of human Dnmt3b [UniProt# Q9UBC3]
Purification Unpurified
Alternative Name DNMT3B (DNMT3B Antibody Abstract)
Background Gene Symbol: DNMT3B
Molecular Weight Theoretical MW: 100 kDa
Gene ID 1789
UniProt Q9UBC3
Application Notes Western Blot 1:1000, Immunohistochemistry 1:200, Immunocytochemistry/Immunofluorescence 1:100-1:1000, Immunoprecipitation 1:1000, Immunohistochemistry-Paraffin 1:200This Dnmt3b antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry on paraffin-embedded sections, Immunoprecipitation and Western Blot, where a band is seen at ~100 kDa. This antibody does not neutralize the enzymatic activity of its corresponding methyltransferase. In ICC/IF, cytoplasmic staining was observed in NIH-3T3 cells. In IHC, predominant nuclear staining was observed, as well as some weaker cytoplasmic signal, in mouse testes. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer ( pH 6.0) is recommended. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol Protocol Specific for NB100-266Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
. Rinse the blot.
. Block the membrane using standard blocking buffer for at least 1 hour.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions.Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.Immunohistochemistry-Paraffin Embedded SectionsAntigen Unmasking:Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.
0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. Staining:
. Wash sections in deionized water three times for 5 minutes each.
. Wash sections in wash buffer for 5 minutes.
. Block each section with 100-400 µL blocking solution for 1 hour at room temperature.
. Remove blocking solution and add 100-400 µL diluted primary antibody. Incubate overnight at 4 degrees C.
. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
. Add 100-400 µL biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
. Add 100-400 µL Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
. Wash sections three times in wash buffer for 5 minutes each.
. Add 100-400 µL DAB substrate to each section and monitor staining closely.
. As soon as the sections develop, immerse slides in deionized water.
. Counterstain sections in hematoxylin.
. Wash sections in deionized water two times for 5 minutes each.
. Dehydrate sections.
. Mount coverslips.Immunocytochemistry ProtocolCulture cells to appropriate density in 35 mm culture dishes or 6-well plates.
. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
Restrictions For Research Use only
Format Liquid
Buffer Whole antisera
Buffer contains: No Preservative
Preservative Without preservative
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C,-20 °C
Storage Comment Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.
Supplier Images
Western Blotting (WB) image for anti-DNMT3B antibody (DNA (Cytosine-5-)-Methyltransferase 3 beta) (ABIN251114) Western blot analysis using ABIN151734 to detect overexpressed DNMT-38 in HCT116 cell...
Immunohistochemistry (IHC) image for anti-DNMT3B antibody (DNA (Cytosine-5-)-Methyltransferase 3 beta) (ABIN251114) Immunohistochemistry: Dnmt3b Antibody [ABIN151734] - Dnmt3b antibody was tested in mo...
Immunofluorescence (IF) image for anti-DNMT3B antibody (DNA (Cytosine-5-)-Methyltransferase 3 beta) (ABIN251114) Immunocytochemistry/Immunofluorescence: Dnmt3b Antibody [ABIN151734] - Dnmt3b antibod...
Product cited in: Liu, Sun, Fong, Yang, Zhang, Yin, Jiang, Liu, Ju, Huang, Bai, Gong, Yan, Zhang, Shao: "An association between overexpression of DNA methyltransferase 3B4 and clear cell renal cell carcinoma." in: Oncotarget, Vol. 8, Issue 12, pp. 19712-19722, 2017 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Human).

Hoang, Horowitz, Chaffee, Qi, Flake, Bruney, Rasor, Rosalez, Wagner, Robinson: "Lens development requires DNMT1 but takes place normally in the absence of both DNMT3A and DNMT3B activity." in: Epigenetics, Vol. 12, Issue 1, pp. 27-40, 2016 (PubMed). Method employed by authors: Western Blotting (WB)

Quist, Jin, Zhu, Smith-Fry, Capecchi, Jones: "The impact of osteoblastic differentiation on osteosarcomagenesis in the mouse." in: Oncogene, Vol. 34, Issue 32, pp. 4278-84, 2015 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Chen, Shabashvili, Nawab, Yang, Dyer, Brown, Hollingshead, Hunter, Kaye, Hochwald, Marquez, Steeg, Zajac-Kaye: "DNA methyltransferase inhibitor, zebularine, delays tumor growth and induces apoptosis in a genetically engineered mouse model of breast cancer." in: Molecular cancer therapeutics, Vol. 11, Issue 2, pp. 370-82, 2012 (PubMed).

Kim, Jung, Lee, Lee, Kang, Lee, Park, Han, Lee, Ramkumar, Sung, Kim: "SIRT1 activation by resveratrol ameliorates cisplatin-induced renal injury through deacetylation of p53." in: American journal of physiology. Renal physiology, Vol. 301, Issue 2, pp. F427-35, 2011 (PubMed).

Kinney, Chin, Vaisvila, Bitinaite, Zheng, Estève, Feng, Stroud, Jacobsen, Pradhan: "Tissue-specific distribution and dynamic changes of 5-hydroxymethylcytosine in mammalian genomes." in: The Journal of biological chemistry, Vol. 286, Issue 28, pp. 24685-93, 2011 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting

Al-Salihi, Yu, Burnett, Alexander, Samlowski, Fitzpatrick: "The depletion of DNA methyltransferase-1 and the epigenetic effects of 5-aza-2'deoxycytidine (decitabine) are differentially regulated by cell cycle progression." in: Epigenetics, Vol. 6, Issue 8, pp. 1021-8, 2011 (PubMed). (Sample species: Human).

Kaminskyy, Surova, Vaculova, Zhivotovsky: "Combined inhibition of DNA methyltransferase and histone deacetylase restores caspase-8 expression and sensitizes SCLC cells to TRAIL." in: Carcinogenesis, Vol. 32, Issue 10, pp. 1450-8, 2011 (PubMed).

Morey Kinney, Smiraglia, James, Moser, Foster, Karpf et al.: "Stage-specific alterations of DNA methyltransferase expression, DNA hypermethylation, and DNA hypomethylation during prostate cancer progression in the transgenic adenocarcinoma of mouse prostate ..." in: Molecular cancer research : MCR, Vol. 6, Issue 8, pp. 1365-74, 2008 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting

Morey, Smiraglia, James, Yu, Moser, Foster, Karpf: "DNA methylation pathway alterations in an autochthonous murine model of prostate cancer." in: Cancer research, Vol. 66, Issue 24, pp. 11659-67, 2006 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting

Xiong, Dowdy, Podratz, Jin, Attewell, Eberhardt, Jiang: "Histone deacetylase inhibitors decrease DNA methyltransferase-3B messenger RNA stability and down-regulate de novo DNA methyltransferase activity in human endometrial cells." in: Cancer research, Vol. 65, Issue 7, pp. 2684-9, 2005 (PubMed). (Sample species: Human). Further details: Western Blotting

Robert, Morin, Beaulieu, Gauthier, Chute, Barsalou, MacLeod: "DNMT1 is required to maintain CpG methylation and aberrant gene silencing in human cancer cells." in: Nature genetics, Vol. 33, Issue 1, pp. 61-5, 2003 (PubMed). (Sample species: Human). Further details: Western Blotting

Weisenberger, Velicescu, Preciado-Lopez, Gonzales, Tsai, Liang, Jones: "Identification and characterization of alternatively spliced variants of DNA methyltransferase 3a in mammalian cells." in: Gene, Vol. 298, Issue 1, pp. 91-9, 2002 (PubMed).

Rhee, Bachman, Park, Jair, Yen, Schuebel, Cui, Feinberg, Lengauer, Kinzler, Baylin, Vogelstein: "DNMT1 and DNMT3b cooperate to silence genes in human cancer cells." in: Nature, Vol. 416, Issue 6880, pp. 552-6, 2002 (PubMed). (Sample species: Human). Further details: Western Blotting

Beaulieu, Morin, Chute, Robert, Nguyen, MacLeod: "An essential role for DNA methyltransferase DNMT3B in cancer cell survival." in: The Journal of biological chemistry, Vol. 277, Issue 31, pp. 28176-81, 2002 (PubMed).

Qiu, Mirkin, Dwivedi: "Differential expression of DNA-methyltransferases in drug resistant murine neuroblastoma cells." in: Cancer detection and prevention, Vol. 26, Issue 6, pp. 444-53, 2002 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunocytochemistry,Western Blotting,Immunofluorescence