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AB269491

Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line

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HSP90AB1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 8 bp deletion, 22 bp deletion; Frameshift: 100%. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

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

4 Images
Western blot - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (AB269491)
  • WB

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (AB269491)

Lanes 1 - 4 : Merged signal (red and green). Green - ab82522 observed at 85 kDa. Red - loading control, ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.

ab82522 was shown to react with Hsp90 beta in wild-type A-431 cells in western blot Loss of signal was observed when HSP90AB1 knockout cell line ab269491 (knockout cell lysate ab269654) was used. Wild-type A-431 and HSP90AB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab82522 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Hsp90 beta antibody [K3705] (<a href='/en-us/products/primary-antibodies/hsp90-beta-antibody-k3705-ab82522'>ab82522</a>) at 1/1000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

HSP90AB1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (ab269491)

Lane 3:

Saos-2 (Human osteosarcoma cell line) whole cell lysate at 20 µg

Lane 4:

HL-60 (Human promyelocytic leukemia cell line) whole cell lysate at 20 µg

Predicted band size: 83 kDa

Observed band size: 85 kDa

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Western blot - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (AB269491)
  • WB

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (AB269491)

Lanes 1 - 4 : Merged signal (red and green). Green - ab82584 observed at 85 kDa. Red - loading control, ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.

ab82584 was shown to react with Hsp90 beta in wild-type A-431 cells in western blot Loss of signal was observed when HSP90AB1 knockout cell line ab269491 (knockout cell lysate ab269654) was used. Wild-type A-431 and HSP90AB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab82584 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Hsp90 beta antibody [K3701] (<a href='/en-us/products/primary-antibodies/hsp90-beta-antibody-k3701-ab82584'>ab82584</a>) at 1/1000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

HSP90AB1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (ab269491)

Lane 3:

Saos-2 (Human osteosarcoma cell line) whole cell lysate at 20 µg

Lane 4:

HL-60 (Human promyelocytic leukemia cell line) whole cell lysate at 20 µg

Predicted band size: 83 kDa

Observed band size: 85 kDa

false

Next Generation Sequencing - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (AB269491)
  • NGS

Supplier Data

Next Generation Sequencing - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (AB269491)

8 bp deletion after Glu172 of the WT protein

Next Generation Sequencing - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (AB269491)
  • NGS

Supplier Data

Next Generation Sequencing - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell line (AB269491)

Knockout achieved by CRISPR/Cas9; X = 8 bp deletion, 22 bp deletion; Frameshift : 100%

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 8 bp deletion, 22 bp deletion; Frameshift: 100%

Disease

Epidermoid Carcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

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.

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What's included?

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Properties and storage information

Gene name
HSP90AB1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Supplementary information

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.
Biological function summary

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.

Pathways

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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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Target data

See full target information HSP90AB1
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Complementary Products

Cell Lines & Lysates

AB269654

Human HSP90AB1 (Hsp90 beta) knockout A-431 cell lysate

Western blot - Human HSP90AB1 (Hsp90 beta) knockout A-431 cell lysate (AB269654)

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Primary Antibodies

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Primary Antibodies

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Proteins & Peptides

AB318253

Recombinant Human HSP90 alpha Protein (His-tag)

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Product promise

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