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AB257254

Human HSP90B1 (GRP94) knockout HEK-293T cell lysate

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HSP90B1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 1.
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Western blot - Human HSP90B1 (GRP94) knockout HEK-293T cell lysate (AB257254)
  • WB

Lab

Western blot - Human HSP90B1 (GRP94) knockout HEK-293T cell lysate (AB257254)

Lane 1 : Wild-type HEK293T cell lysate (20 μg)

Lane 2 : HSP90B1 knockout HEK293T cell lysate (20 μg)

Lane 3 : HeLa cell lysate (20 μg)

Lanes 1-3 : Merged signal (red and green). Green - ab238126 observed at 94 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab238126 Anti-GRP94 antibody [EPR22847-50] was shown to specifically react with GRP94 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266313 (knockout cell lysate ab257254) was used. Wild-type and GRP94 knockout samples were subjected to SDS-PAGE. ab238126 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GRP94 antibody [EPR22847-50] (<a href='/en-us/products/primary-antibodies/grp94-antibody-epr22847-50-ab238126'>ab238126</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

HSP90B1 knockout HEK293T cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Predicted band size: 92 kDa

Observed band size: 94 kDa

false

Western blot - Human HSP90B1 (GRP94) knockout HEK-293T cell lysate (AB257254)
  • WB

Lab

Western blot - Human HSP90B1 (GRP94) knockout HEK-293T cell lysate (AB257254)

Lane 1 : Wild-type HEK293T cell lysate (20 μg)

Lane 2 : HSP90B1 knockout HEK293T cell lysate (20 μg)

Lane 3 : HeLa cell lysate (20 μg)

Lanes 1-3 : Merged signal (red and green). Green - ab108606 observed at 94 kDa. Red - loading control ab8245 observed at 36 kDa.

ab108606 Anti-GRP94 antibody [EPR3988] was shown to specifically react with GRP94 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266313 (knockout cell lysate ab257254) was used. Wild-type and GRP94 knockout samples were subjected to SDS-PAGE. ab108606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-GRP94 antibody [EPR3988] (<a href='/en-us/products/primary-antibodies/grp94-antibody-epr3988-ab108606'>ab108606</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

HSP90B1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human HSP90B1 (GRP94) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hsp90b1-grp94-knockout-hek-293t-cell-line-ab266313'>ab266313</a>)

Lane 3:

HeLa 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: 92 kDa

Observed band size: 94 kDa

false

Sanger Sequencing - Human HSP90B1 (GRP94) knockout HEK-293T cell lysate (AB257254)
  • Sanger seq

Unknown

Sanger Sequencing - Human HSP90B1 (GRP94) knockout HEK-293T cell lysate (AB257254)

Homozygous : 10 bp deletion in exon 1

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, Homozygous: 10 bp deletion in exon 1.

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Complementary Products

Primary Antibodies

AB238126

Anti-GRP94 antibody [EPR22847-50]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] (AB238126)

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Recombinant human VEGF 165A protein (Active)

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Mass Spectrometry - Recombinant human VEGF 165A protein (Active) (AB259412)

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Proteins & Peptides

AB52101

Recombinant human Cleaved Caspase-3 protein (Active)

Western blot - Recombinant human Cleaved Caspase-3 protein (Active) (AB52101)

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Proteins & Peptides

AB87144

Recombinant Human GRP94 protein

Western blot - Recombinant Human GRP94 protein (AB87144)

  • 0
  • 0
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Reactivity data

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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?

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Properties and storage information

Gene name
HSP90B1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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.

GRP94 also known as HSP90b1 is a glycoprotein that belongs to the heat shock protein 90 family. It has a molecular mass of approximately 94 kDa. GRP94 is expressed mainly in the endoplasmic reticulum and appears in high levels in cells undergoing stress. This protein acts as a chaperone assisting in the proper folding and assembly of other proteins playing an essential role in protein homeostasis. GRP94 ensures the quality control of proteins by preventing misfolding and aggregation.
Biological function summary

GRP94 influences the stability of many secretory and cell-surface proteins. It forms part of a multi-protein complex that stabilizes client proteins and assists their proper folding. GRP94 is essential for the maturation of proteins involved in the immune response including immunoglobulins and integrins. Overexpression of GRP94 has been noted in various cancers where it supports the folding of proteins required for tumor growth and survival.

Pathways

GRP94 is an important element of the protein folding quality control mechanism within the endoplasmic reticulum. It functions in coordination with other chaperones like BiP and calnexin within the unfolded protein response (UPR) pathway. The UPR pathway is essential during stress conditions where increased protein synthesis occurs. GRP94 also contributes to calcium homeostasis and directly interacts with proteins like integrins affecting cell adhesion and motility.

GRP94 has been implicated in the progression of cancer and neurodegenerative diseases. Overexpression of GRP94 is often linked to cancer where it stabilizes oncogenic proteins necessary for cancer cell survival. In neurodegenerative disorders GRP94 shows altered expression levels which may influence the pathogenesis of diseases like Alzheimer's. The interaction between GRP94 and other proteins such as tau protein in Alzheimer's highlights its potential role in disease mechanisms.
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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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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'.

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