Human PGAM5 knockout HeLa cell line
- Advanced Validation
- What is this?
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PGAM5 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
View Alternative Names
BXLBv68, Bcl-XL-binding protein v68, MGC5352, PGAM5_HUMAN, Phosphoglycerate mutase family member 5, Serine/threonine protein phosphatase PGAM5 mitochondrial, Serine/threonine-protein phosphatase PGAM5, mitochondrial
- WB
Lab
Western blot - Human PGAM5 knockout HeLa cell line (AB265141)
ab244218 Anti-PGAM5 antibody [CL0624] was shown to specifically react with PGAM5 in HeLa wild-type cells (ab255928). Loss of signal was observed when knockout cell line ab265141 (knockout cell lysate ab257581) was used. Wild-type and PGAM5 knockout samples were subjected to SDS-PAGE. ab244218 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4° at 1 μg/ml 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-PGAM5 antibody [CL0624] (<a href='/en-us/products/primary-antibodies/pgam5-antibody-cl0624-ab244218'>ab244218</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa lysate at 20 µg
Lane 2:
PGAM5 knockout HeLa lysate at 20 µg
Lane 2:
Western blot - Human PGAM5 knockout HeLa cell line (ab265141)
Lane 3:
HepG2 lysate at 20 µg
Lane 4:
Daudi lysate at 20 µg
Predicted band size: 32 kDa
Observed band size: 32 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PGAM5 knockout HeLa cell line (AB265141)
Homozygous : 1 bp insertion in exon 1.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human PGAM5 knockout HeLa cell line (AB265141)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PGAM5 KO HeLa (PGAM5 knockout human cervical adenocarcinoma epithelial cell) (ab265141) cells labelling PGAM5 with ab322207 at 1/100 (4.85 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ 1000 2ug/ml dilution (Green).
Confocal image showing mitochondria staining in parental HeLa cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ 1000 2ug/ml dilution.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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
Zygosity
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
PGAM5 impacts pathways related to cellular stress responses and energy metabolism. It participates in programmed cell death and modulates mitochondrial dynamics. It can serve as part of a complex with other proteins adjusting the balance between mitophagy and apoptosis. Through interaction with key signaling molecules PGAM5 bridges various biochemical processes relevant to cell survival and energy utilization.
Pathways
Its function integrates into mitophagy and necroptosis. PGAM5 collaborates with proteins like PINK1 and PARKIN contributing to the clearance of damaged mitochondria via the mitophagy pathway. In necroptosis PGAM5 interacts with RIPK1 and RIPK3 regulating necrotic cell death. These interactions position PGAM5 at the crossroads of pathways critical to maintaining cellular homeostasis and response to stress.
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
AB308448
Anti-PGAM5 (Long isoform) antibody [EPR27145-21] - BSA and Azide free
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] - BSA and Azide free (AB308448)](https://content.abcam.com/products/images/pgam5-long-isoform-antibody-epr27145-21-bsa-and-azide-free-ab308448--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img409825.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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