Human MTAP knockout HeLa cell pellet (ab279112)
Overview
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Product name
Human MTAP knockout HeLa cell pellet
See all MTAP 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 insertion in exon1 and Insertion of the selection cassette in exon1. -
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 MTAP 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
Plays a major role in polyamine metabolism and is important for the salvage of both adenine and methionine. -
Tissue specificity
Ubiquitously expressed. -
Sequence similarities
Belongs to the PNP/MTAP phosphorylase family. -
Cellular localization
Cytoplasm. - Information by UniProt
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Alternative names
- 5' methylthioadenosine phosphorylase
- 5''-methylthioadenosine phosphorylase
- BDMF
see all
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit Human MTAP 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 MTAP knockout HeLa cell pellet (ab279112)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: MTAP knockout HeLa cell lysate (20 µg)
Lane 3: HT-29 cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab254265 observed at 32 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab254265 Anti-MTAP antibody [EPR22570-76] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265272 (knockout cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab254265 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 10000 dilution for 1 hour at room temperature before imaging.
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Western blot - Human MTAP knockout HeLa cell pellet (ab279112)Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: MTAP knockout HeLa cell lysate (20 µg)
Lane 3: HT-29 cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
anes 1- 4: Merged signal (red and green). Green - ab126770 observed at 32 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab126770 Anti-MTAP antibody [EPR6893] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265272 (knockout cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab126770 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 10000 dilution for 1 hour at room temperature before imaging.
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Sanger Sequencing - Human MTAP knockout HeLa cell pellet (ab279112)Allele-1: 1 bp insertion in exon1
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Sanger Sequencing - Human MTAP knockout HeLa cell pellet (ab279112)Allele-2: Insertion of the selection cassette in exon1
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Sanger Sequencing - Human MTAP knockout HeLa cell pellet (ab279112)Allele-3: Insertion of the selection cassette in exon1