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.
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
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.
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.
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.
Hsp90 alpha also known as Hsp90AA1 or 5G5 is a molecular chaperone with an approximate molecular weight of 90 kDa. It plays an important role in the folding stability and function of many client proteins. This protein is expressed ubiquitously in eukaryotic cells with high levels in the cytoplasm and the endoplasmic reticulum. Hsp90 alpha operates through a unique ATPase-driven chaperone cycle that modulates protein conformations helping in the assembly and disassembly of protein complexes.
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.
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.
Hsp90 alpha is linked to cancer and neurodegenerative diseases. Its enhanced expression in tumors correlates with the stabilization of oncogenic client proteins such as HER2 contributing to cancer progression. Additionally Hsp90 alpha relates to disorders like Alzheimer's disease due to its role in handling misfolded proteins. The protein interacts with co-chaperones like H1L1 which may influence its impact on disease outcomes highlighting the therapeutic potential of targeting Hsp90 alpha in these conditions.
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Anti-Hsp90 antibody [D7a] 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 Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell lysate ab258458) was used. Wild-type and Hsp90 knockout samples were subjected to SDS-PAGE. Anti-Hsp90 antibody [D7a] ab59459 and Anti-GAPDH antibody[EPR16891] - Loading Control (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 (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (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] (Anti-Hsp90 antibody [D7a] ab59459) 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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Predicted band size: 85 kDa
Observed band size: 90 kDa
Anti-Hsp90 alpha antibody [2G5.G3] 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 Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell lysate ab258458) was used. Wild-type and Hsp90 alpha knockout samples were subjected to SDS-PAGE. Anti-Hsp90 alpha antibody [2G5.G3] ab79849 and Anti-GAPDH antibody[EPR16891] - Loading Control (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 (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (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] (Anti-Hsp90 alpha antibody [2G5.G3] ab79849) 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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Predicted band size: 85 kDa
Observed band size: 90 kDa
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.
Homozygous: 1 bp insertion in exon3
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