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DNA (Cytosine-5-)-Methyltransferase 3 beta (DNMT3B) (AA 4-101) antibody

Details for Product No. ABIN251114
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Antigen
Synonyms ICF, ICF1, M.HsaIIIB, cb91, dnmt3bl, dnmt7, sb:cb91, wu:fb16h07, cb633, dnmt3b, DNMT3B, LOC100218113
Epitope
AA 4-101
(8), (7), (6), (2), (2), (2), (1), (1), (1), (1), (1), (1)
Reactivity
Human, Mouse (Murine)
(71), (26), (20), (1), (1), (1), (1)
Host
Rabbit
(63), (9), (4)
Clonality
Polyclonal
Conjugate
Un-conjugated
(2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Western Blotting (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
(60), (27), (23), (17), (14), (12), (12), (8), (7), (3), (2), (2)
Pubmed 7 references available
Quantity 25 mL
Shipping to United States (Change)
Catalog No. ABIN251114
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Immunogen Amino acids 4-101 of human Dnmt3b
Purification Whole antisera
Alternative Name DNMT3B
Background Inactivation of tumor suppressor genes is a key component of the development of allcommon forms of human cancers. Aberrant methylation may contribute to many of thechanges, both physiological and pathological, associated with aging, including tumordevelopment. Hypermethylation of CpG island-containing promoters is associated withthe inactivation of genes important to tumor angiogenesis and progression. DNAmethytransferase (DNMT3b) is necessary for the maintenance of the methylationprocess. Alternate Names: anti-ICF antibody, anti-M.HsaIIIB antibody.
Gene Symbol: DNMT3B
Gene ID 1789
UniProt Q9UBC3
Application Notes This 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.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:100-1:1000, Immunohistochemistry 1:200, Immunohistochemistry-Paraffin 1:200, immunoprecipitation 1:1000, Western Blot 1:1000
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
Preservative Without preservative
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C
Storage Comment 4 °C short term. Aliquot and store at -20 °C long term.
Supplier Images
anti-DNA (Cytosine-5-)-Methyltransferase 3 beta (DNMT3B) (AA 4-101) antibody Western blot analysis using ABIN151734 to detect overexpressed DNMT-38 in HCT116 cells. Lane 1: 40 ug of nuclear extract from untransfected T24 cells. Lane 2-3: 40 ug of nuclear extracts transiently transfected to overexpress DNMT3B2 and DNMT3B3, respectively.
General Rhee, Bachman, Park et al.: "DNMT1 and DNMT3b cooperate to silence genes in human cancer cells." in: Nature, Vol. 416, Issue 6880, pp. 552-6, 2002 (PubMed).

Beaulieu, Morin, Chute et al.: "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).

Weisenberger, Velicescu, Preciado-Lopez et al.: "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).

Robert, Morin, Beaulieu et al.: "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).

Xiong, Dowdy, Podratz et al.: "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).

Morey, Smiraglia, James et al.: "DNA methylation pathway alterations in an autochthonous murine model of prostate cancer." in: Cancer research, Vol. 66, Issue 24, pp. 11659-67, 2006 (PubMed).

Morey Kinney, Smiraglia, James 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 model." in: Molecular cancer research : MCR, Vol. 6, Issue 8, pp. 1365-74, 2008 (PubMed).

Hosts (63), (9), (4)
Reactivities (71), (26), (20), (1), (1), (1), (1)
Applications (60), (27), (23), (17), (14), (12), (12), (8), (7), (3), (2), (2)
Conjugates (2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Epitopes (8), (7), (6), (2), (2), (2), (1), (1), (1), (1), (1), (1)
Request Want additional data for this product?

The Independent Validation Initiative strives to provide you with high quality data. Find out more

Catalog No. ABIN251114
Contact our Customer Service for availability and price in your country.
Add to Basket

Order hotline:

  • +1 404 474 4654
  • +1 888 205 9894 (TF)
Validation Images
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