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BAG2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.

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Images

Western blot - Human BAG2 knockout HeLa cell line (AB265907), expandable thumbnail
  • Sanger Sequencing - Human BAG2 knockout HeLa cell line (AB265907), expandable thumbnail
  • Sanger Sequencing - Human BAG2 knockout HeLa cell line (AB265907), expandable thumbnail

Key facts

Cell type
HeLa
Species or organism
Human
Tissue
Cervix
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, 2 bp insertion in exon 1 and Insertion of the selection cassette in exon 1

Alternative names

BAG-family molecular chaperone regulator-2, BAG2_HUMAN, Bcl-2-associated athanogene 2, KIAA0576, MGC149462, OTTHUMP00000016668, dJ417I1.2, dJ417I1.2 (BAG family molecular chaperone regulator 2)

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BAG2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.

Key facts

Cell type
HeLa
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, 2 bp insertion in exon 1 and Insertion of the selection cassette in exon 1
Disease
Adenocarcinoma
Concentration
Loading...

Properties

Gene name
BAG2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot

Quality control

STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female

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.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The BAG2 protein also known as Bcl-2-associated athanogene 2 is a co-chaperone that modulates the activity of heat shock proteins especially Hsp70. It has a molecular mass of approximately 27 kDa. BAG2 regulates the degradation of misfolded proteins by binding to the ATPase domain of Hsp70. It is expressed widely in human tissues including the brain heart and skeletal muscle suggesting its broad physiological importance.

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.

Associated diseases and disorders

BAG2 has been linked to neurodegenerative diseases such as Alzheimer's disease. The dysregulation of BAG2 expression and function could lead to the accumulation of misfolded proteins exacerbating neurodegeneration. BAG2 also associates with various cancers where its expression may impact tumor survival and resistance to stress. Its interaction with Hsp70 and Bcl-2 proteins plays a role in these pathologies influencing disease progression and therapeutic outcomes.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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