Human PMS2 knockout HeLa cell line
- Advanced Validation
- What is this?
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PMS2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 2 and 2 bp deletion in exon 2 and Insertion of the selection cassette in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
DNA mismatch repair gene homologue, DNA mismatch repair protein PMS2, HNPCC4, H_DJ0042M02.9, MLH4, Mismatch repair endonuclease PMS2, Mismatch repair gene PMSL2, PMS1 homolog 2 mismatch repair system, PMS1 protein homolog 2, PMS2 postmeiotic segregation increased 2, PMS2 postmeiotic segregation increased 2 (S. cerevisiae), PMS2CL, PMS2_HUMAN, PMSL2, Postmeiotic segregation increased, S. cerevisiae, 2
- WB
Lab
Western blot - Human PMS2 knockout HeLa cell line (AB261776)
Lanes 1- 2 : Merged signal (red and green). Green - ab110638 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab110638 was shown to react with PMS2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261776 (knockout cell lysate ab257142) was used. Wild-type HeLa and PMS2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab110638 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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.
All lanes:
Western blot - Anti-PMS2 antibody [EPR3947] (<a href='/en-us/products/primary-antibodies/pms2-antibody-epr3947-ab110638'>ab110638</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human PMS2 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-pms2-knockout-hela-cell-lysate-ab257142'>ab257142</a>) at 20 µg
Secondary
Lanes 1 - 2:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Lanes 1 - 2:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 96 kDa
Observed band size: 120 kDa,37 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human PMS2 knockout HeLa cell line (AB261776)
Allele-3 : Insertion of the selection cassette in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human PMS2 knockout HeLa cell line (AB261776)
Allele-1 : 16 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human PMS2 knockout HeLa cell line (AB261776)
Allele-2 : 2 bp deletion in exon 2.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
PMS2 operates as part of the DNA mismatch repair (MMR) complex. It collaborates with other MutL homologs including MLH1 to form a heterodimer which is essential for repairing DNA replication errors. It safeguards genomic integrity and prevents mutations from accumulating in dividing cells serving important functions in cellular viability and genetic stability.
Pathways
PMS2 is involved in the DNA damage response and cell cycle regulation. The protein plays a vital role in the mismatch repair (MMR) pathway. PMS2 partners primarily with MLH1 within this pathway and both proteins work in conjunction to recognize and initiate repair on erroneous DNA sequences that emerge during replication preventing illegitimate recombination and chromosomal rearrangements.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB110638
BOND RX™ Validated|RabMAb|Recombinant|KO Validated|20ul selling size|Trial Size|Trial Size|Trial Size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMS2 antibody [EPR3947] (AB110638)](https://content.abcam.com/products/images/pms2-antibody-epr3947-ab110638--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img186217.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com