Human RRM2B (p53R2) knockout HCT116 cell line
- Advanced Validation
- What is this?
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RRM2B KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.
View Alternative Names
DKFZp686M05248, MGC102856, MGC42116, MTDPS8A, MTDPS8B, P53 inducible ribonucleotide reductase small subunit 2 short form beta, RIR2B_HUMAN, RRM 2B, Ribonucleoside diphosphate reductase M2 subunit B, Ribonucleoside-diphosphate reductase subunit M2 B, Ribonucleotide reductase M2 B, Ribonucleotide reductase M2 B (TP53 inducible), Ribonucleotide reductase small subunit like 2 p53 inducible, TP53-inducible ribonucleotide reductase M2 B, p53 R2, p53 inducible ribonucleotide reductase small subunit 2 homolog, p53-inducible ribonucleotide reductase small subunit 2-like protein
- WB
Lab
Western blot - Human RRM2B (p53R2) knockout HCT116 cell line (AB266897)
Lanes 1- 2 : Merged signal (red and green). Green - ab154194 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab154194 was shown to react with p53R2 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266897 (knockout cell lysate ab257216) was used. Wild-Type HCT116 and RRM2B knockout HCT116 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. ab154194 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° 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-p53R2 antibody [EPR8816] (<a href='/en-us/products/primary-antibodies/p53r2-antibody-epr8816-ab154194'>ab154194</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT116 cell lysate at 20 µg
Lane 2:
RRM2B knockout HCT116 cell lysate at 20 µg
Lane 2:
Western blot - Human RRM2B (p53R2) knockout HCT116 cell line (ab266897)
Predicted band size: 40 kDa
Observed band size: 40 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RRM2B (p53R2) knockout HCT116 cell line (AB266897)
Allele-1 : 1 bp insertion in exon1
- Sanger seq
Unknown
Sanger Sequencing - Human RRM2B (p53R2) knockout HCT116 cell line (AB266897)
Allele-2 : Insertion of the selection cassette in exon 1.
Reactivity data
Product details
Recommended control: Human wild-type HCT116 cell line (ab255451). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
McCoY5a + 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
P53R2 plays a role in the synthesis of deoxyribonucleotide triphosphates (dNTPs) for DNA repair. It forms part of a ribonucleotide reductase complex working with other subunits to ensure cells have sufficient dNTP pools for DNA replication and repair. The presence of p53R2 is integral for DNA damage response pathways where it gets activated to supply dNTPs during periods when the p53 tumor suppressor pathway senses DNA damage and activates DNA repair processes.
Pathways
P53R2 is involved in DNA damage response and cell cycle regulation. It interacts with the tumor suppressor protein p53 especially during instances of genotoxic stress facilitating DNA repair and cell cycle arrest to prevent propagation of damaged DNA. Additionally p53R2 relates to proteins such as p21 which are components of the p53 pathway influencing cell cycle arrest and allowing time for DNA repair before cell division resumes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
Primary Antibodies
AB154194
BOND RX™ Validated|RabMAb|Recombinant|KO Validated|Trial Size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] (AB154194)](https://content.abcam.com/products/images/p53r2-antibody-epr8816-ab154194--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img102274.jpg)
- 0
- 0
![Western blot - Anti-Thymidylate Synthase antibody [EPR4545] (AB108995)](https://content.abcam.com/products/images/thymidylate-synthase-antibody-epr4545-ab108995--western-blot-img116051.jpg)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RRM2 antibody [EPR11820] (AB172476)](https://content.abcam.com/products/images/rrm2-antibody-epr11820-ab172476--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img422250.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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