HSP90AB1 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: 5 bp deletion in exon 3.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 3
90 kda heat shock protein beta HSP90 beta, D6S182, FLJ26984, HS90B_HUMAN, HSP 84, HSP 90, HSP 90 b, HSP90 BETA, HSP90AB1, HSPC2, HSPCB, Heat shock 84 kDa, Heat shock 90kD protein 1, beta, Heat shock 90kDa protein 1 beta, Heat shock protein 90 alpha family class B member 1, Heat shock protein 90 kDa, Heat shock protein 90kDa alpha (cytosolic) class B member 1, Heat shock protein 90kDa alpha family class B member 1, Heat shock protein HSP 90-beta, Heat shock protein beta
HSP90AB1 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: 5 bp deletion in exon 3.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 3
HSP90AB1
Knockout
CRISPR technology
Sanger Sequencing, Western blot
Homozygous
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
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.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T 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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
This supplementary information is collated from multiple sources and compiled automatically.
Hsp90 beta also known as Hsp90AB1 or Hsp90 protein is a heat shock protein of approximately 90 kDa. It is a molecular chaperone found in most eukaryotic cells. Hsp90 beta helps in the proper folding stabilization and degradation of many proteins. Unlike its isoform Hsp90 alpha Hsp90 beta has a more stable expression and is not typically induced by stress. This protein is primarily localized in the cytosol but can also be present in other cellular compartments depending on the cellular state.
Hsp90 beta functions to maintain protein homeostasis and cellular integrity. It forms part of a multi-protein chaperone complex which includes cochaperones such as Hop Hsp70 and p23 necessary for its full functionality. Hsp90 beta supports the maturation of steroid hormone receptors kinases and other client proteins. It plays an important role in the cell cycle regulation through its interaction with various proteins ensuring proper cell division and growth.
Hsp90 beta is deeply involved in signal transduction and cellular stress response pathways. Its interaction with the Akt pathway is significant for cell survival signals. Hsp90 beta also participates in the MAP kinase pathway affecting cell growth and differentiation. The protein associates with multiple kinases including RAF and Src which are important for downstream signaling.
The dysregulation of Hsp90 beta is associated with cancer and neurodegenerative diseases. In cancer Hsp90 beta stabilizes many oncoproteins making it a potential therapeutic target for inhibiting tumor growth. It interacts with client proteins like the proto-oncogene c-Src promoting tumorigenesis. In neurodegenerative disorders such as Alzheimer's disease improper interaction between Hsp90 beta and tau proteins can contribute to disease progression impacting neuronal function.
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Lanes 1 - 4: Merged signal (red and green). Green - Anti-Hsp90 beta antibody [EPR16621] ab203085 observed at 85 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-Hsp90 beta antibody [EPR16621] ab203085 was shown to react with Hsp90 beta in wild-type HEK-293T cells in western blot with loss of signal observed in HSP90ab1 knockout cell line ab266117 (HSP90ab1 knockout cell lysate Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate ab257190). Wild-type and HSP90ab1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-Hsp90 beta antibody [EPR16621] ab203085 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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-Hsp90 beta antibody [EPR16621] (Anti-Hsp90 beta antibody [EPR16621] ab203085) at 1/5000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HSP90AB1 knockout HEK-293T cell lysate at 20 µg
Lane 3: Saos-2 cell lysate at 20 µg
Lane 4: HL-60 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 85 kDa
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Hsp90 antibody [H90-10] ab58950 observed at 85 kDa. Red - loading control Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
Anti-Hsp90 antibody [H90-10] ab58950 was shown to react with Hsp90 in wild-type HEK-293T cells in western blot with loss of signal observed in HSP90ab1 knockout cell line ab266117 (HSP90ab1 knockout cell lysate Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate ab257190). Wild-type and HSP90ab1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-Hsp90 antibody [H90-10] ab58950 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4° at 0.5 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Hsp90 antibody [H90-10] (Anti-Hsp90 antibody [H90-10] ab58950) at 0.5 µg/mL
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HSP90AB1 knockout HEK-293T cell lysate at 20 µg
Lane 3: Saos-2 cell lysate at 20 µg
Lane 4: HL-60 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa, 85 kDa
Observed band size: 85 kDa
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Hsp90 beta antibody [E296] ab32568 observed at 85 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-Hsp90 beta antibody [E296] ab32568 was shown to react with Hsp90 beta in wild-type HEK-293T cells in western blot with loss of signal observed in HSP90ab1 knockout cell line ab266117 (HSP90ab1 knockout cell lysate Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate ab257190). Wild-type and HSP90ab1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-Hsp90 beta antibody [E296] ab32568 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 200000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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-Hsp90 beta antibody [E296] (Anti-Hsp90 beta antibody [E296] ab32568) at 1/200000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HSP90AB1 knockout HEK-293T cell lysate at 20 µg
Lane 3: Saos-2 cell lysate at 20 µg
Lane 4: HL-60 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 85 kDa
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Hsp90 beta antibody [H90-10] ab53497 observed at 85 kDa. Red - loading control Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
Anti-Hsp90 beta antibody [H90-10] ab53497 was shown to react with Hsp90 beta in wild-type HEK-293T cells in western blot with loss of signal observed in HSP90ab1 knockout cell line ab266117 (HSP90ab1 knockout cell lysate Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate ab257190). Wild-type and HSP90ab1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-Hsp90 beta antibody [H90-10] ab53497 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4° at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Hsp90 beta antibody [H90-10] (Anti-Hsp90 beta antibody [H90-10] ab53497) at 1/5000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HSP90AB1 knockout HEK-293T cell lysate at 20 µg
Lane 3: Saos-2 cell lysate at 20 µg
Lane 4: HL-60 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 85 kDa
Lanes 1-4: Merged signal (red and green). Green - Anti-Hsp90 beta antibody [E296] ab32568 observed at 90 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-Hsp90 beta antibody [E296] ab32568 Anti-Hsp90 beta antibody [E296] was shown to specifically react with Hsp90 beta in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266117 (knockout cell lysate Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate ab257190) was used. Wild-type and Hsp90 beta knockout samples were subjected to SDS-PAGE. Anti-Hsp90 beta antibody [E296] ab32568 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-Hsp90 beta antibody [E296] (Anti-Hsp90 beta antibody [E296] ab32568) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: HSP90AB1 knockout HEK293T cell lysate at 20 µg
Lane 3: Jurkat cell lysate at 20 µg
Lane 4: SH-SY5Y cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 83 kDa
Observed band size: 90 kDa
This data was developed using the same antibody clone in a different buffer formulation (Anti-Hsp90 beta antibody [E296] ab32568).
Lanes 1-4: Merged signal (red and green). Green - Anti-Hsp90 beta antibody [E296] ab32568 observed at 90 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-Hsp90 beta antibody [E296] ab32568 Anti-Hsp90 beta antibody [E296] was shown to specifically react with Hsp90 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266117 (knockout cell lysate Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate ab257190) was used. Wild-type and Hsp90 knockout samples were subjected to SDS-PAGE. Anti-Hsp90 beta antibody [E296] ab32568 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.
Lanes 1-4: Merged signal (red and green). Green - Anti-Hsp90 beta antibody [EPR16621] ab203085 observed at 90 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-Hsp90 beta antibody [EPR16621] ab203085 Anti-Hsp90 beta antibody [EPR16621] was shown to specifically react with Hsp90 beta in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266117 (knockout cell lysate Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate ab257190) was used. Wild-type and Hsp90 beta knockout samples were subjected to SDS-PAGE. Anti-Hsp90 beta antibody [EPR16621] ab203085 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-Hsp90 beta antibody [EPR16621] (Anti-Hsp90 beta antibody [EPR16621] ab203085) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: HSP90AB1 knockout HEK293T cell lysate at 20 µg
Lane 3: Jurkat cell lysate at 20 µg
Lane 4: SH-SY5Y cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 83 kDa
Observed band size: 90 kDa
This data was developed using the same antibody clone in a different buffer formulation (Anti-Hsp90 beta antibody [EPR16621] ab203085).
Lanes 1-4: Merged signal (red and green). Green - Anti-Hsp90 beta antibody [EPR16621] ab203085 observed at 90 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-Hsp90 beta antibody [EPR16621] ab203085 Anti-Hsp90 beta antibody [EPR16621] was shown to specifically react with Hsp90 beta in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266117 (knockout cell lysate Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate ab257190) was used. Wild-type and Hsp90 beta knockout samples were subjected to SDS-PAGE. Anti-Hsp90 beta antibody [EPR16621] ab203085 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.
Representative images HSP90AB1 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: 5 bp deletion in exon3
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