Human BAG2 knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human BAG2 knockout HeLa cell line (AB265907)
Lanes 1-3 : Merged signal (red and green). Green - ab79406 observed at 25 kDa. Red - loading control ab8245 observed at 36 kDa.
ab79406 Anti-BAG2 antibody [EPR3567] was shown to specifically react with BAG2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265907 (knockout cell lysate ab257369) was used. Wild-type and BAG2 knockout samples were subjected to SDS-PAGE. ab79406 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-BAG2 antibody [EPR3567] (<a href='/en-us/products/primary-antibodies/bag2-antibody-epr3567-ab79406'>ab79406</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
BAG2 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human BAG2 knockout HeLa cell line (ab265907)
Lane 3:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
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 1/10000 dilution
Predicted band size: 23 kDa
Observed band size: 25 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human BAG2 knockout HeLa cell line (AB265907)
Allele-1 : 2 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human BAG2 knockout HeLa cell line (AB265907)
Allele-2 : Insertion of the selection cassette in exon 1.
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BAG2 antibody [EPR3567] (AB79406)](https://content.abcam.com/products/images/bag2-antibody-epr3567-ab79406--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img107859.jpg)
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
The role of BAG2 involves maintaining protein homeostasis a critical aspect of cellular functions. It forms a complex with Hsp70 assisting in the proper folding of newly synthesized proteins and preventing their aggregation. BAG2 also impacts the ubiquitin-proteasome system by blocking the association of Hsp70 with CHIP an E3 ubiquitin ligase. This regulation prevents the proteolytic degradation of client proteins.
Pathways
BAG2 functions within the cellular stress response and protein folding pathways. It has a significant involvement in the Hsp70-mediated protein folding pathway where it influences the fate of various substrate proteins. The interaction of BAG2 with Hsp70 and the ubiquitin-proteasome pathway highlights its contribution to cellular homeostasis. Additionally BAG2's modulation of these pathways suggests a link with apoptosis regulation where it associates indirectly with Bcl-2 family members.
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'.
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