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Rabbit Recombinant Monoclonal Hsc70 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 51 publications.


Images

Immunoprecipitation - Anti-Hsc70 antibody [EP1531Y] (AB51052), expandable thumbnail
  • Western blot - Anti-Hsc70 antibody [EP1531Y] (AB51052), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Hsc70 antibody [EP1531Y] (AB51052), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Hsc70 antibody [EP1531Y] (AB51052), expandable thumbnail
  • Western blot - Anti-Hsc70 antibody [EP1531Y] (AB51052), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Tested
Expected
Expected
Rat
Expected
Expected
Tested
Expected
Expected

Tested
Tested

Species
Human
Dilution info
1/50
Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human
Dilution info
1/10 - 1/20
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/500 - 1/5000
Notes

-

Species
Rat
Dilution info
1/500 - 1/5000
Notes

-

Species
Human
Dilution info
1/500 - 1/5000
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100 - 1/250
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human
Dilution info
1/30 - 1/70
Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Associated Products

Select an associated product type

3 products for Alternative Product

Target data

Function

Molecular chaperone implicated in a wide variety of cellular processes, including protection of the proteome from stress, folding and transport of newly synthesized polypeptides, chaperone-mediated autophagy, activation of proteolysis of misfolded proteins, formation and dissociation of protein complexes, and antigen presentation. Plays a pivotal role in the protein quality control system, ensuring the correct folding of proteins, the re-folding of misfolded proteins and controlling the targeting of proteins for subsequent degradation (PubMed:21148293, PubMed:21150129, PubMed:23018488, PubMed:24732912, PubMed:27916661, PubMed:2799391, PubMed:36586411). This is achieved through cycles of ATP binding, ATP hydrolysis and ADP release, mediated by co-chaperones (PubMed:12526792, PubMed:21148293, PubMed:21150129, PubMed:23018488, PubMed:24732912, PubMed:27916661). The co-chaperones have been shown to not only regulate different steps of the ATPase cycle of HSP70, but they also have an individual specificity such that one co-chaperone may promote folding of a substrate while another may promote degradation (PubMed:12526792, PubMed:21148293, PubMed:21150129, PubMed:23018488, PubMed:24732912, PubMed:27916661). The affinity of HSP70 for polypeptides is regulated by its nucleotide bound state. In the ATP-bound form, it has a low affinity for substrate proteins. However, upon hydrolysis of the ATP to ADP, it undergoes a conformational change that increases its affinity for substrate proteins. HSP70 goes through repeated cycles of ATP hydrolysis and nucleotide exchange, which permits cycles of substrate binding and release. The HSP70-associated co-chaperones are of three types: J-domain co-chaperones HSP40s (stimulate ATPase hydrolysis by HSP70), the nucleotide exchange factors (NEF) such as BAG1/2/3 (facilitate conversion of HSP70 from the ADP-bound to the ATP-bound state thereby promoting substrate release), and the TPR domain chaperones such as HOPX and STUB1 (PubMed:24121476, PubMed:24318877, PubMed:26865365, PubMed:27474739). Plays a critical role in mitochondrial import, delivers preproteins to the mitochondrial import receptor TOMM70 (PubMed:12526792). Acts as a repressor of transcriptional activation. Inhibits the transcriptional coactivator activity of CITED1 on Smad-mediated transcription. Component of the PRP19-CDC5L complex that forms an integral part of the spliceosome and is required for activating pre-mRNA splicing. May have a scaffolding role in the spliceosome assembly as it contacts all other components of the core complex. Binds bacterial lipopolysaccharide (LPS) and mediates LPS-induced inflammatory response, including TNF secretion by monocytes (PubMed:10722728, PubMed:11276205). Substrate recognition component in chaperone-mediated autophagy (CMA), a selective protein degradation process that mediates degradation of proteins with a -KFERQ motif: HSPA8/HSC70 specifically recognizes and binds cytosolic proteins bearing a -KFERQ motif and promotes their recruitment to the surface of the lysosome where they bind to lysosomal protein LAMP2 (PubMed:11559757, PubMed:2799391, PubMed:36586411). KFERQ motif-containing proteins are eventually transported into the lysosomal lumen where they are degraded (PubMed:11559757, PubMed:2799391, PubMed:36586411). In conjunction with LAMP2, facilitates MHC class II presentation of cytoplasmic antigens by guiding antigens to the lysosomal membrane for interaction with LAMP2 which then elicits MHC class II presentation of peptides to the cell membrane (PubMed:15894275). Participates in the ER-associated degradation (ERAD) quality control pathway in conjunction with J domain-containing co-chaperones and the E3 ligase STUB1 (PubMed:23990462). It is recruited to clathrin-coated vesicles through its interaction with DNAJC6 leading to activation of HSPA8/HSC70 ATPase activity and therefore uncoating of clathrin-coated vesicles (By similarity).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal Hsc70 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 51 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EP1531Y
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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7 product images

  • Immunoprecipitation - Anti-Hsc70 antibody [EP1531Y] (ab51052), expandable thumbnail

    Immunoprecipitation - Anti-Hsc70 antibody [EP1531Y] (ab51052)

    ab51052 (purified) at 1/20 immunoprecipitating Hsc70 in 10 μg HeLa (Lanes 1 and 2, observed at 71 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP Detection Reagent (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

    All lanes: Immunoprecipitation - Anti-Hsc70 antibody [EP1531Y] (ab51052)

    Predicted band size: 70 kDa

  • Western blot - Anti-Hsc70 antibody [EP1531Y] (ab51052), expandable thumbnail

    Western blot - Anti-Hsc70 antibody [EP1531Y] (ab51052)

    Lanes 1- 4: Merged signal (red and green). Green - ab51052 observed at 71 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    ab51052 was shown to react with Hsc70 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human HSPA8 (Hsc70) knockout HeLa cell line ab265664 (knockout cell lysate Human HSPA8 (Hsc70) knockout HeLa cell lysate ab256944) was used. Wild-type HeLa and HSPA8 knockout HeLa cell lysates 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 (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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] (ab51052) 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 (Human HSPA8 (Hsc70) knockout HeLa cell line ab265664)

    Lane 3: A431 cell lysate at 20 µg

    Lane 4: MCF7 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 70 kDa

    Observed band size: 71 kDa

  • Flow Cytometry (Intracellular) - Anti-Hsc70 antibody [EP1531Y] (ab51052), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Hsc70 antibody [EP1531Y] (ab51052)

    Overlay histogram showing MCF7 cells fixed in 4% PFA and stained with purified ab51052 at a dilution of 1 in 70 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).

  • Immunocytochemistry/ Immunofluorescence - Anti-Hsc70 antibody [EP1531Y] (ab51052), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Hsc70 antibody [EP1531Y] (ab51052)

    Immunofluorescence staining of MCF7 cells with purified ab51052 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab51052 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.

  • Western blot - Anti-Hsc70 antibody [EP1531Y] (ab51052), expandable thumbnail

    Western blot - Anti-Hsc70 antibody [EP1531Y] (ab51052)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Hsc70 antibody [EP1531Y] (ab51052) at 1/5000 dilution

    Lane 1: C6 cell lysate at 10 µg

    Lane 2: PC-12 cell lysate at 10 µg

    Lane 3: Mouse heart tissue lysate at 10 µg

    Lane 4: Rat heart tissue lysate at 10 µg

    Secondary

    All lanes: HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 70 kDa

    Observed band size: 71 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsc70 antibody [EP1531Y] (ab51052), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsc70 antibody [EP1531Y] (ab51052)

    Immunohistochemical staining of paraffin embedded human cerebral cortex with purified ab51052 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Western blot - Anti-Hsc70 antibody [EP1531Y] (ab51052), expandable thumbnail

    Western blot - Anti-Hsc70 antibody [EP1531Y] (ab51052)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-Hsc70 antibody [EP1531Y] (ab51052) at 1/5000 dilution

    Lane 1: human fetal brain lysate at 10 µg

    Lane 2: HeLa cell lysate at 10 µg

    Lane 3: HEK293 cell lysate at 10 µg

    Secondary

    All lanes: HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 70 kDa

    Observed band size: 71 kDa

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