Human RIPK2 (RIP2) knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human RIPK2 (RIP2) knockout HeLa cell line (AB264688)
Lanes 1-4 : Merged signal (red and green). Green - ab75257 observed at 65 kDa. Red - loading control ab181602 observed at 37 kDa.
ab75257 Anti-RIP2 antibody [AF28D3] was shown to specifically react with RIP2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264688 (knockout cell lysate ab258636) was used. Wild-type and RIP2 knockout samples were subjected to SDS-PAGE. ab75257 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4° at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-RIP2 antibody [AF28D3] (<a href='/en-us/products/primary-antibodies/rip2-antibody-af28d3-ab75257'>ab75257</a>) at 1/2000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
RIPK2 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human RIPK2 (RIP2) knockout HeLa cell line (ab264688)
Lane 3:
Ramos cell lysate at 20 µg
Lane 4:
Jurkat cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution
Predicted band size: 61 kDa
Observed band size: 65 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RIPK2 (RIP2) knockout HeLa cell line (AB264688)
Allele-1 : 2 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human RIPK2 (RIP2) knockout HeLa cell line (AB264688)
Allele-2 : 1 bp insertion in exon 1.
Complementary Products
Cell Lines & Lysates
AB261782
Human IFNAR1 (Interferon alpha/beta receptor 1) knockout HeLa cell line
- 0
- 0
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
RIP2 interacts with key components of the immune signaling network. It forms part of a complex with other proteins like nucleotide-binding oligomerization domain (NOD) proteins influencing immune responses. By binding to NOD1 and NOD2 RIP2 modulates inflammatory signaling pathways leading to the activation of NF-kB and the production of inflammatory cytokines. This protein helps regulate the body's defense against bacterial infections and influences inflammation processes.
Pathways
RIP2 plays an integral role in the NOD-like receptor signaling pathway and is also involved in toll-like receptor signaling. In its pathways RIP2 interacts closely with proteins like IKK complexes and the Tak1 complex orchestrating signaling cascades leading to the activation of NF-kB and MAPK pathways. These pathways are essential for instigating the immune response and managing inflammatory signals highlighting how RIP2 acts as a pivotal mediator in these biological routes.
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
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