Human HSPA8 (Hsc70) knockout HeLa cell line
- Advanced Validation
- What is this?
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(2 Publications)
HSPA8 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 2 and Insertion of the selection cassette in exon 2.
View Alternative Names
2410008N15Rik, Constitutive heat shock protein 70, Epididymis luminal protein 33, Epididymis secretory sperm binding protein Li 72p, HEL 33, HEL S 72p, HSC54, HSC71, HSP71, HSP73, HSP7C_HUMAN, HSPA10, HSPA8, Heat shock 70 kDa protein 8, Heat shock 70kD protein 10, Heat shock 70kD protein 8, Heat shock cognate 71 kDa protein, Heat shock cognate protein 54, Heat shock cognate protein 71 kDa, Heat shock protein 8, Heat shock protein A8, Heat shock protein family A (Hsp70) member 8, Heat-shock70-KD protein 10, formerly, Hsc73, LAP 1, LPS associated protein, LPS associated protein 1, Lipopolysaccharide associated protein 1, MGC102007, MGC106514, MGC114311, MGC118485, MGC131511, MGC29929, N-myristoyltransferase inhibitor protein 71, NIP71
- WB
Lab
Western blot - Human HSPA8 (Hsc70) knockout HeLa cell line (AB265664)
Lanes 1- 4 : Merged signal (red and green). Green - ab51052 observed at 71 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab51052 was shown to react with Hsc70 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265664 (knockout cell lysate ab256944) was used. Wild-type HeLa and HSPA8 knockout HeLa 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. ab51052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 500 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-Hsc70 antibody [EP1531Y] (<a href='/en-us/products/primary-antibodies/hsc70-antibody-ep1531y-ab51052'>ab51052</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
HSPA8 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human HSPA8 (Hsc70) knockout HeLa cell line (ab265664)
Lane 3:
A431 cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
Predicted band size: 70 kDa
Observed band size: 71 kDa
false
- WB
Lab
Western blot - Human HSPA8 (Hsc70) knockout HeLa cell line (AB265664)
Lanes 1- 4 : Merged signal (red and green). Green - ab112549 observed at 70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab112549 was shown to react with Hsc70 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265664 (knockout cell lysate ab256944) was used. Wild-type HeLa and HSPA8 knockout HeLa 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. ab112549 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 μg/ml 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-Hsc70 antibody (<a href='/en-us/products/primary-antibodies/hsc70-antibody-ab112549'>ab112549</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
HSPA8 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human HSPA8 (Hsc70) knockout HeLa cell line (ab265664)
Lane 3:
A431 cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
Predicted band size: 70 kDa
Observed band size: 70 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human HSPA8 (Hsc70) knockout HeLa cell line (AB265664)
Allele-1 : 1 bp insertion in exon 2.
- Cell Culture
Unknown
Cell Culture - Human HSPA8 (Hsc70) knockout HeLa cell line (AB265664)
Representative images of HSPA8 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human HSPA8 (Hsc70) knockout HeLa cell line (AB265664)
Allele-2 : Insertion of the selection cassette in exon 2.
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
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
Hsc70 participates in several processes beyond protein folding. This includes its involvement in the transport of proteins across cellular membranes and the degradation of misfolded proteins. Hsc70 often works in conjunction with co-chaperones and forms part of large protein complexes such as the chaperone machinery involved in clathrin-mediated endocytosis. It also plays a part in cellular stress responses supporting cell survival under various stress conditions.
Pathways
Hsc70 integrates into fundamental cellular pathways such as the ubiquitin-proteasome system and the ER-associated degradation pathway. These pathways are important for protein quality control where Hsc70 collaborates with other heat shock proteins like Hsp40 to facilitate the correct folding and clearance of proteins. In the context of autophagy Hsc70 mediates the recognition and trafficking of specific substrates to lysosomes for degradation demonstrating its importance in maintaining cellular health through these pathways.
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
Publications (2)
Recent publications for all applications. Explore the full list and refine your search
American journal of physiology. Gastrointestinal and liver physiology 327:G267-G283 PubMed38860860
2024
Applications
Unspecified application
Species
Unspecified reactive species
Acta biochimica et biophysica Sinica 56:356-365 PubMed38419499
2024
Applications
Unspecified application
Species
Unspecified reactive species
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Product promise
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