Human POLB (DNA Polymerase beta) knockout HeLa cell pellet (ab279011)
Overview
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Product name
Human POLB (DNA Polymerase beta) knockout HeLa cell pellet
See all DNA Polymerase beta 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, 5 bp deletion in exon 1 and Insertion of the selection cassette in exon 1. -
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 POLB 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
Repair polymerase. Conducts "gap-filling" DNA synthesis in a stepwise distributive fashion rather than in a processive fashion as for other DNA polymerases. Has a 5'-deoxyribose-5-phosphate lyase (dRP lyase) activity. -
Sequence similarities
Belongs to the DNA polymerase type-X family. -
Domain
Residues 239-252 form a flexible loop which appears to affect the polymerase fidelity. -
Post-translational
modificationsMethylation by PRMT6 stimulates the polymerase activity by enhancing DNA binding and processivity. -
Cellular localization
Nucleus, Cytoplasm - Information by UniProt
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Alternative names
- DNA directed DNA polymerase beta
- DNA pol beta
- DNA polymerase beta
see all
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit Human POLB 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 POLB (DNA Polymerase beta) knockout HeLa cell pellet (ab279011)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: POLB knockout HeLa cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lanes 1-3: Merged signal (red and green). Green - ab175197 observed at 38 kDa. Red - loading control, ab7291 observed at 50 kDa.
ab175197 Anti-DNA Polymerase beta antibody [EPR12722(B)] was shown to specifically react with DNA Polymerase beta in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261748 (knockout cell lysate ab257096) was used. Wild-type and DNA Polymerase beta knockout samples were subjected to SDS-PAGE. ab175197 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 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 POLB (DNA Polymerase beta) knockout HeLa cell pellet (ab279011)Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: POLB knockout HeLa cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lanes 1-3: Merged signal (red and green). Green - ab26343 observed at 38 kDa. Red - loading control, ab7291 observed at 50 kDa.
ab26343 Anti-DNA Polymerase beta antibody was shown to specifically react with DNA Polymerase beta in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261748 (knockout cell lysate ab257096) was used. Wild-type and DNA Polymerase beta knockout samples were subjected to SDS-PAGE. ab26343 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 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 POLB (DNA Polymerase beta) knockout HeLa cell pellet (ab279011)Allele-1: 5 bp deletion in exon 1; Allele-2: Insertion of the selection cassette in exon 1
