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

Details for Product anti-DNMT3B Antibody No. ABIN251114, Supplier: Log in to see
Antigen
  • cb91
  • cb633
  • DNMT3B
  • dnmt3b
  • dnmt3bl
  • dnmt7
  • ICF
  • ICF1
  • LOC100218113
  • M.HsaIIIB
  • MmuIIIB
  • sb:cb91
  • wu:fb16h07
Reactivity
Human, Mouse (Murine)
108
48
18
2
1
1
1
1
Host
Rabbit
79
30
2
2
1
Clonality
Polyclonal
Conjugate
This DNMT3B antibody is un-conjugated
4
4
3
3
2
2
2
2
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)
97
55
36
33
30
29
26
23
11
5
3
2
2
1
1
1
1
Supplier
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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 1:200
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 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...
Product cited in: 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