#32578 DNMT3A (D2H4B) Rabbit mAb
|内在性||130, 95, 85||Rabbit IgG||-20℃|
|DNMT3A (D2H4B) Rabbit mAb recognizes endogenous levels of total DNMT3A protein. This antibody detects multiple isoforms of DNMT3A, including isoform 1 and isoform 2. The antibody does not cross-react with DNMT3B or other DNMT proteins.|
|Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro233 of human DNMT3A protein.|
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Western blot analysis of extracts from NCCIT, NTERA-2 cl.D1, and HCT 116 cells using DNMT3A (D2H4B) Rabbit mAb. This antibody detect multiple isoforms of DNMT3A, including isoforms 1 and 2.
Immunoprecipitation of DNMT3A from NCCIT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is DNMT3A (D2H4B) Rabbit mAb. Western blot analysis was performed using DNMT3A (E9P2F) Rabbit mAb #49768.
Confocal immunofluorescent analysis of NCCIT (left, high-expressing) and 293T (right, low-expressing) cells using DNMT3A (D2H4B) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). According to WB analysis, 293T cells preferentially express the larger DNMT3A isoform 1, while NCCIT cells also express high levels of two smaller isoforms, including isoform 2.
Flow cytometric analysis of PC-3 cells (blue) and NCCIT cells (green) using DNMT3A (D2H4B) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either DNMT3A (E9P2F) Rabbit mAb #49768 or DNMT3A (D2H4B) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figures show binding across HUWE1, a known target gene of DNMT3A (see additional figures containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either DNMT3A (D2H4B) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HUWE1 Intron 26 Control Primers #33770, human ARHGEF2 intron 13 primers, human SDHAP1 intron 10 primers, and SimpleChIP® Human GAPDH Exon 1 Primers #5516. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Methylation of DNA at cytosine residues in mammalian cells is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting and development (1,2). Three families of mammalian DNA methyltransferases have been identified: DNMT1, DNMT2 and DNMT3 (1,2). DNMT1 is constitutively expressed in proliferating cells and functions as a maintenance methyltransferase, transferring proper methylation patterns to newly synthesized DNA during replication. DNMT3A and DNMT3B are strongly expressed in embryonic stem cells with reduced expression in adult somatic tissues. DNMT3A and DNMT3B function as de novo methyltransferases that methylate previously unmethylated regions of DNA. DNMT2 is expressed at low levels in adult somatic tissues and its inactivation affects neither de novo nor maintenance DNA methylation. DNMT1, DNMT3A and DNMT3B together form a protein complex that interacts with histone deacetylases (HDAC1, HDAC2, Sin3A), transcriptional repressor proteins (RB, TAZ-1) and heterochromatin proteins (HP1, SUV39H1), to maintain proper levels of DNA methylation and facilitate gene silencing (3-8). Improper DNA methylation contributes to diseased states such as cancer (1,2). Hypermethylation of promoter CpG islands within tumor suppressor genes correlates with gene silencing and the development of cancer. In addition, hypomethylation of bulk genomic DNA correlates with and may contribute to the onset of cancer. DNMT1, DNMT3A and DNMT3B are over-expressed in many cancers, including acute and chronic myelogenous leukemias, in addition to colon, breast and stomach carcinomas (9-12).
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Illumina is a registered trademark of Illumina, Inc.
Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.