AB266117

Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line

Advanced Validation
What is this?

Be the first to review this product! Submit a review

(0 Publication)

HSP90AB1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

8 Images

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Lanes 1 - 4 : Merged signal (red and green). Green - ab203085 observed at 85 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

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 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 ab203085 and 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 (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 [EPR16621] (<a href='/en-us/products/primary-antibodies/hsp90-beta-antibody-epr16621-ab203085'>ab203085</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (ab266117)

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

Predicted band size: 83 kDa

Observed band size: 85 kDa

false

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Lanes 1-4 : Merged signal (red and green). Green - ab203085 observed at 90 kDa. Red - loading control ab8245 observed at 36 kDa.

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 ab257190) was used. Wild-type and Hsp90 beta knockout samples were subjected to SDS-PAGE. ab203085 and 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 (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-Hsp90 beta antibody [EPR16621] (<a href='/en-us/products/primary-antibodies/hsp90-beta-antibody-epr16621-ab203085'>ab203085</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-hsp90ab1-hsp90-beta-knockout-hek-293t-cell-lysate-ab257190'>ab257190</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 83 kDa

Observed band size: 90 kDa

false

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Lanes 1-4 : Merged signal (red and green). Green - ab32568 observed at 90 kDa. Red - loading control ab8245 observed at 36 kDa.

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 ab257190) was used. Wild-type and Hsp90 beta knockout samples were subjected to SDS-PAGE. ab32568 and 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 (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-Hsp90 beta antibody [E296] (<a href='/en-us/products/primary-antibodies/hsp90-beta-antibody-e296-ab32568'>ab32568</a>) 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 2:

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (ab266117)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Secondary

All lanes:

Predicted band size: 83 kDa

Observed band size: 90 kDa

false

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Lanes 1 - 4 : Merged signal (red and green). Green - ab32568 observed at 85 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

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 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 ab32568 and 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 (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 [E296] (<a href='/en-us/products/primary-antibodies/hsp90-beta-antibody-e296-ab32568'>ab32568</a>) 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 2:

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (ab266117)

Lane 3:

Saos-2 cell lysate at 20 µg

Lane 4:

HL-60 cell lysate at 20 µg

Predicted band size: 83 kDa

Observed band size: 85 kDa

false

Unknown

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Lanes 1 - 4 : Merged signal (red and green). Green - ab53497 observed at 85 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.

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 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 ab53497 and 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (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] (<a href='/en-us/products/primary-antibodies/hsp90-beta-antibody-h90-10-ab53497'>ab53497</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-hsp90ab1-hsp90-beta-knockout-hek-293t-cell-lysate-ab257190'>ab257190</a>) at 20 µg

Lane 3:

Saos-2 cell lysate at 20 µg

Lane 4:

HL-60 cell lysate at 20 µg

Predicted band size: 83 kDa

Observed band size: 85 kDa

false

Unknown

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Lanes 1 - 4 : Merged signal (red and green). Green - ab58950 observed at 85 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.

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 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 ab58950 and 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (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] (<a href='/en-us/products/primary-antibodies/hsp90-antibody-h90-10-ab58950'>ab58950</a>) 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 2:

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (ab266117)

Lane 3:

Saos-2 cell lysate at 20 µg

Lane 4:

HL-60 cell lysate at 20 µg

Predicted band size: 83 kDa,85 kDa

Observed band size: 85 kDa

false

Sanger Sequencing - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Sanger seq

Unknown

Sanger Sequencing - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Homozygous : 5 bp deletion in exon3

Cell Culture - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Cell Culture

Lab

Cell Culture - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

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.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 3

style

left-column

style

left-column

Reactivity data

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

style

left-column

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.

style

left-column

What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab266117-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266117 Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line", "number":"AB266117-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab255449 Human wild-type HEK-293T cell line", "number":"AB266117-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab266117-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266117 Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line", "number":"AB266117-CMP01", "productcode":"" } ] } } }

style

left-column

Properties and storage information

Gene name

HSP90AB1

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

style

left-column

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.

style

left-column

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.

style

left-column

style

left-column

Quality control

STR analysis

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

style

left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style

left-column

Product protocols

Visit the General protocols
Visit the Troubleshooting

style

left-column

Target data

See full target information HSP90AB1

style

left-column

style

left-column

style

right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Sanger Sequencing - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Cell Culture - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (AB266117)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Complementary Products

Cell Lines & Lysates

Primary Antibodies

Primary Antibodies

Proteins & Peptides

Reactivity data

Product details

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

Subculture guidelines

Culture medium

Cryopreservation medium

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Target data

Product promise