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AB256944

Human HSPA8 (Hsc70) knockout HeLa cell lysate

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HSPA8 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.

View Alternative Names

2410008N15Rik, Constitutive heat shock protein 70, Epididymis luminal protein 33, Epididymis secretory sperm binding protein Li 72p, HEL 33, HEL S 72p, HSC54, HSC71, HSP71, HSP73, HSP7C_HUMAN, HSPA10, HSPA8, Heat shock 70 kDa protein 8, Heat shock 70kD protein 10, Heat shock 70kD protein 8, Heat shock cognate 71 kDa protein, Heat shock cognate protein 54, Heat shock cognate protein 71 kDa, Heat shock protein 8, Heat shock protein A8, Heat shock protein family A (Hsp70) member 8, Heat-shock70-KD protein 10, formerly, Hsc73, LAP 1, LPS associated protein, LPS associated protein 1, Lipopolysaccharide associated protein 1, MGC102007, MGC106514, MGC114311, MGC118485, MGC131511, MGC29929, N-myristoyltransferase inhibitor protein 71, NIP71

4 Images
Western blot - Human HSPA8 (Hsc70) knockout HeLa cell lysate (AB256944)
  • WB

Lab

Western blot - Human HSPA8 (Hsc70) knockout HeLa cell lysate (AB256944)

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

Lane 2 : HSPA8 knockout HeLa cell lysate (20µg)

Lane 3 : A431 cell lysate (20µg)

Lane 4 : MCF7 cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab51052 observed at 71 kDa. Red - loading control ab8245 observed at 37 kDa.

ab51052 Rabbit monoclonal [EP1531Y] to Hsc70 was shown to specifically react with Hsc70 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265664 (knockout cell lysate ab256944) was used. Wild-type and Hsc70 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab51052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-Hsc70 antibody [EP1531Y] (<a href='/en-us/products/primary-antibodies/hsc70-antibody-ep1531y-ab51052'>ab51052</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

HSPA8 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human HSPA8 (Hsc70) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-hspa8-hsc70-knockout-hela-cell-line-ab265664'>ab265664</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 70 kDa

Observed band size: 71 kDa

false

Western blot - Human HSPA8 (Hsc70) knockout HeLa cell lysate (AB256944)
  • WB

Lab

Western blot - Human HSPA8 (Hsc70) knockout HeLa cell lysate (AB256944)

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

Lane 2 : HSPA8 knockout HeLa cell lysate (20µg)

Lane 3 : A431 cell lysate (20µg)

Lane 4 : MCF7 cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab112549 observed at 70 kDa. Red - loading control ab8245 observed at 37 kDa.

ab112549 Rabbit polyclonal to Hsc70 was shown to specifically react with Hsc70 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265664 (knockout cell lysate ab256944) was used. Wild-type and Hsc70 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab112549 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 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-Hsc70 antibody (<a href='/en-us/products/primary-antibodies/hsc70-antibody-ab112549'>ab112549</a>) at 1 µg/mL

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

HSPA8 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human HSPA8 (Hsc70) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-hspa8-hsc70-knockout-hela-cell-line-ab265664'>ab265664</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 70 kDa

Observed band size: 70 kDa

false

Sanger Sequencing - Human HSPA8 (Hsc70) knockout HeLa cell lysate (AB256944)
  • Sanger seq

Unknown

Sanger Sequencing - Human HSPA8 (Hsc70) knockout HeLa cell lysate (AB256944)

Allele-1 : 1 bp insertion in exon2

Sanger Sequencing - Human HSPA8 (Hsc70) knockout HeLa cell lysate (AB256944)
  • Sanger seq

Unknown

Sanger Sequencing - Human HSPA8 (Hsc70) knockout HeLa cell lysate (AB256944)

Allele-2 : Insertion of the selection cassette in exon2

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.

Disease

Adenocarcinoma

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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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
HSPA8
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.

The heat shock cognate 70 (Hsc70) protein also known as HSPA8 plays an important role in chaperone-mediated cellular processes. It exhibits a molecular weight of approximately 70 kDa. Hsc70 is abundantly expressed in various tissues and organs including the brain muscle and liver. As a member of the heat shock protein family Hsc70 maintains protein homeostasis by preventing protein aggregation and assisting in protein folding.
Biological function summary

Hsc70 participates in several processes beyond protein folding. This includes its involvement in the transport of proteins across cellular membranes and the degradation of misfolded proteins. Hsc70 often works in conjunction with co-chaperones and forms part of large protein complexes such as the chaperone machinery involved in clathrin-mediated endocytosis. It also plays a part in cellular stress responses supporting cell survival under various stress conditions.

Pathways

Hsc70 integrates into fundamental cellular pathways such as the ubiquitin-proteasome system and the ER-associated degradation pathway. These pathways are important for protein quality control where Hsc70 collaborates with other heat shock proteins like Hsp40 to facilitate the correct folding and clearance of proteins. In the context of autophagy Hsc70 mediates the recognition and trafficking of specific substrates to lysosomes for degradation demonstrating its importance in maintaining cellular health through these pathways.

Hsc70 shows a strong connection to neurodegenerative diseases and cancer. Misregulation of Hsc70 activity can result in protein aggregation seen in disorders such as Parkinson's disease. This situation often links Hsc70 to proteins like alpha-synuclein which accumulate abnormally in these diseases. In cancer altered expression of Hsc70 can promote tumor development by hyper-accommodating the stress conditions within the tumor microenvironment. Hsc70's interplay with proteins such as p53 highlights its role in the pathology of tumorigenesis.
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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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Complementary Products

Primary Antibodies

AB51052

Anti-Hsc70 antibody [EP1531Y]

RabMAb|Recombinant|KO Validated

Western blot - Anti-Hsc70 antibody [EP1531Y] (AB51052)

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  • 2
Proteins & Peptides

AB316845

Recombinant Human HSPA8 Protein (His-tag)

  • 0
  • 0
Primary Antibodies

AB52649

Anti-GM130 antibody [EP892Y] - cis-Golgi Marker

RabMAb|Recombinant|Lab Essentials

Immunoprecipitation - Anti-GM130 antibody [EP892Y] - cis-Golgi Marker (AB52649)

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Cell Lines & Lysates

AB265664

Human HSPA8 (Hsc70) knockout HeLa cell line

Advanced Validation

Western blot - Human HSPA8 (Hsc70) knockout HeLa cell line (AB265664)

  • 0
  • 0

Product promise

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For full details, please see our Terms & Conditions

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