Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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HSP90AA1 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 3.
View Alternative Names
EL52, HS90A_HUMAN, HSP 86, HSP 90, HSP89A, HSP90A, HSP90AA1, HSP90ALPHA, HSP90N, HSPC1, HSPCA, HSPCAL1, HSPCAL4, HSPN, Heat shock 86 kDa, Heat shock 90kD protein 1, alpha like 4, Heat shock 90kDa protein 1 alpha, Heat shock protein 90kDa alpha (cytosolic) class A member 1, Heat shock protein HSP 90-alpha, Hsp89, LAP 2, LPS-associated protein 2, Renal carcinoma antigen NY-REN-38, epididymis luminal secretory protein 52, heat shock 90kD protein 1, alpha, heat shock 90kD protein, alpha-like 4, lipopolysaccharide-associated protein 2
- WB
Lab
Western blot - Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell line (AB266591)
Lanes 1-4 : Merged signal (red and green). Green - ab79849 observed at 90 kDa. Red - loading control ab181602 observed at 36 kDa.
ab79849 Anti-Hsp90 alpha antibody [2G5.G3] was shown to specifically react with Hsp90 alpha in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266591 (knockout cell lysate ab258458) was used. Wild-type and Hsp90 alpha knockout samples were subjected to SDS-PAGE. ab79849 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) 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® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Hsp90 alpha antibody [2G5.G3] (<a href='/en-us/products/primary-antibodies/hsp90-alpha-antibody-2g5g3-ab79849'>ab79849</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
HSP90AA1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell line (ab266591)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
HAP1 whole cell lyate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution
Predicted band size: 85 kDa
Observed band size: 90 kDa
false
- WB
Unknown
Western blot - Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell line (AB266591)
Lanes 1-4 : Merged signal (red and green). Green - ab59459 observed at 90 kDa. Red - loading control ab181602 observed at 36 kDa.
ab59459 Anti-Hsp90 antibody [D7a] was shown to specifically react with Hsp90 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266591 (knockout cell lysate ab258458) was used. Wild-type and Hsp90 knockout samples were subjected to SDS-PAGE. ab59459 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) 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® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Hsp90 antibody [D7a] (<a href='/en-us/products/primary-antibodies/hsp90-antibody-d7a-ab59459'>ab59459</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
HSP90AA1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell line (ab266591)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
HAP1 whole cell lyate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution
Predicted band size: 85 kDa
Observed band size: 90 kDa
false
- Cell Culture
Lab
Cell Culture - Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell line (AB266591)
Representative images HSP90AA1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T 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 M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell line (AB266591)
Homozygous : 1 bp insertion in exon3
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). 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
Hsp90 alpha assists in maintaining protein homeostasis and is part of a larger chaperome complex including co-chaperones and other chaperones such as Hsp70. It plays an important role in stress response by stabilizing proteins and preventing aggregation. Hsp90 alpha is essential for the normal function of several kinases and transcription factors which are important for cell signaling and regulation processes. It functions as a mediator for cellular response to environmental cues.
Pathways
Hsp90 alpha is an integral component in the signal transduction and cell cycle control pathways. It interacts with various signaling proteins and receptors like AKT and steroid hormone receptors to facilitate their proper folding and activation. Furthermore Hsp90 alpha influences the MAPK/ERK pathway playing a role in cell proliferation and differentiation. Its interaction with Hsp90 protein clients creates a regulatory framework across multiple pathways allowing precise control over cellular function.
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
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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