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AB257190

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

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HSP90AB1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 5 bp deletion in exon3.

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

7 Images
Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)
  • WB

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)

Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : HSP90AB1 knockout HEK-293T cell lysate 20 ug
Lane 3 : Saos-2 cell lysate 20 ug
Lane 4 : HL-60 cell lysate 20 ug
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°C 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 (<a href='/en-us/products/cell-lines/human-hsp90ab1-hsp90-beta-knockout-hek-293t-cell-line-ab266117'>ab266117</a>)

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

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)
  • WB

Unknown

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)

Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : HSP90AB1 knockout HEK-293T cell lysate 20 ug
Lane 3 : Saos-2 cell lysate 20 ug
Lane 4 : HL-60 cell lysate 20 ug
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°C 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:

HSP90AB1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hsp90ab1-hsp90-beta-knockout-hek-293t-cell-line-ab266117'>ab266117</a>)

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

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)
  • WB

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)

Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : HSP90AB1 knockout HEK-293T cell lysate 20 ug
Lane 3 : Saos-2 cell lysate 20 ug
Lane 4 : HL-60 cell lysate 20 ug
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°C 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 (<a href='/en-us/products/cell-lines/human-hsp90ab1-hsp90-beta-knockout-hek-293t-cell-line-ab266117'>ab266117</a>)

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

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)
  • WB

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : HSP90AB1 knockout HEK293T cell lysate (20 ug)
Lane 3 : Jurkat cell lysate (20 ug)
Lane 4 : SH-SY5Y cell lysate (20 ug)

ab203085 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 line (<a href='/en-us/products/cell-lines/human-hsp90ab1-hsp90-beta-knockout-hek-293t-cell-line-ab266117'>ab266117</a>)

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

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

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)
  • WB

Lab

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : HSP90AB1 knockout HEK293T cell lysate (20 ug)
Lane 3 : Jurkat cell lysate (20 ug)
Lane 4 : SH-SY5Y cell lysate (20 ug)

ab32568 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 (<a href='/en-us/products/cell-lines/human-hsp90ab1-hsp90-beta-knockout-hek-293t-cell-line-ab266117'>ab266117</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

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)
  • WB

Unknown

Western blot - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)

Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : HSP90AB1 knockout HEK-293T cell lysate 20 ug
Lane 3 : Saos-2 cell lysate 20 ug
Lane 4 : HL-60 cell lysate 20 ug
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°C at 0.5 ug/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 (<a href='/en-us/products/cell-lines/human-hsp90ab1-hsp90-beta-knockout-hek-293t-cell-line-ab266117'>ab266117</a>)

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 lysate (AB257190)
  • Sanger seq

Unknown

Sanger Sequencing - Human HSP90AB1 (Hsp90 beta) knockout HEK-293T cell lysate (AB257190)

Homozygous : 5 bp deletion in exon3

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 5 bp deletion in exon3.

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Reactivity data

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

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257190-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human HSP90AB1 knockout HEK293T cell lysate", "number":"AB257190-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB257190-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
HSP90AB1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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

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Product promise

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