Human DNMT3A knockout HeLa cell pellet (ab278817)
Overview
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Product name
Human DNMT3A knockout HeLa cell pellet
See all Dnmt3a kits -
Product overview
Abcam’s knockout cell pellets give you access to native proteins, without the need to culture cells. Our knockout cell pellets are prepared from our single-gene knockout cell lines and provide an additional offering to our cell lysates.
Cells are snap-frozen to provide high quality pellets that are suitable for extraction with alternative lysis buffers or for preparation of lysates from subcellular fractions. Our knockout cell pellets are suitable for a variety of applications, including PCR, gene expression profiling and DNA library preparation. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon4 and 1 bp insertion in exon4. -
Passage number
Knockout validation
Sanger Sequencing, Western Blot (WB)Notes
Pellet size: 5 million cells/vial.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit Human DNMT3A knockout HeLa cell pellet 1 vial Human wild-type HeLa cell pellet 1 vial -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
Target
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Function
Required for genome wide de novo methylation and is essential for the establishment of DNA methylation patterns during development. DNA methylation is coordinated with methylation of histones. It modifies DNA in a non-processive manner and also methylates non-CpG sites. May preferentially methylate DNA linker between 2 nucleosomal cores and is inhibited by histone H1. Plays a role in paternal and maternal imprinting. Required for methylation of most imprinted loci in germ cells. Acts as a transcriptional corepressor for ZNF238. Can actively repress transcription through the recruitment of HDAC activity. -
Tissue specificity
Highly expressed in fetal tissues, skeletal muscle, heart, peripheral blood mononuclear cells, kidney, and at lower levels in placenta, brain, liver, colon, spleen, small intestine and lung. -
Sequence similarities
Belongs to the C5-methyltransferase family.
Contains 1 ADD domain.
Contains 1 GATA-type zinc finger.
Contains 1 PHD-type zinc finger.
Contains 1 PWWP domain. -
Domain
The PWWP domain is essential for targeting to pericentric heterochromatin. -
Post-translational
modificationsSumoylated; sumoylation disrupts the ability to interact with histone deacetylases (HDAC1 and HDAC2) and repress transcription. -
Cellular localization
Nucleus. Cytoplasm. Accumulates in the major satellite repeats at pericentric heterochromatin. - Information by UniProt
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Alternative names
- DNA (cytosine 5) methyltransferase 3 alpha
- DNA (cytosine 5) methyltransferase 3A
- DNA (cytosine-5)-methyltransferase 3A
see all
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit Human DNMT3A knockout HeLa cell pellet 1 vial Human wild-type HeLa cell pellet 1 vial -
Research areas
-
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
Images
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Western blot - Human DNMT3A knockout HeLa cell pellet (ab278817)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: DNMT3A knockout HeLa cell lysate (20 µg)
Lane 3: HEK-293 cell lysate (20 µg)
Lane 4: Daudi cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab227823 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.
ab227823 Anti-Dnmt3a antibody [EPR18455-94] was shown to specifically react with Dnmt3a in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261793 (knockout cell lysate ab257128) was used. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab227823 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Human DNMT3A knockout HeLa cell pellet (ab278817)Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: DNMT3A knockout HeLa cell lysate (20 µg)
Lane 3: HEK-293 cell lysate (20 µg)
Lane 4: Daudi cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab188470 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.
ab188470 Anti-Dnmt3a antibody [EPR18455] was shown to specifically react with Dnmt3a in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261793 (knockout cell lysate ab257128) was used. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab188470 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Sanger Sequencing - Human DNMT3A knockout HeLa cell pellet (ab278817)Allele-1: 1 bp deletion in exon4
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Sanger Sequencing - Human DNMT3A knockout HeLa cell pellet (ab278817)Allele-2: 1 bp insertion in exon4