HSPA8 Monoclonal Antibody
Description
Definition and Target Specificity
HSPA8 Monoclonal Antibody is a laboratory-generated immunoglobulin that selectively binds to the HSPA8 protein (Heat Shock Protein Family A Member 8), also known as HSC70. This constitutively expressed chaperone regulates protein folding, degradation via chaperone-mediated autophagy (CMA), and stress adaptation . The antibody is produced using hybridoma technology, ensuring high specificity for human, mouse, and rat HSPA8 isoforms .
Production and Validation
The development process involves:
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Immunization: Mice are immunized with recombinant human HSPA8 protein (amino acids 2–646) or synthesized peptides .
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Hybridoma Generation: Splenocytes from immunized mice are fused with myeloma cells to create antibody-producing hybridomas .
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Purification: Antibodies are isolated using Protein A/G affinity chromatography, achieving >95% purity .
Protein Localization and Stress Response
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Subcellular Tracking: HSPA8 monoclonal antibodies have identified nuclear translocation of HSPA8 during heat shock or oxidative stress, essential for cell survival .
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Autophagy Regulation: Studies using these antibodies demonstrated HSPA8’s role in forming liquid-liquid phase separation (LLPS) complexes with RHOB and BECN1, enhancing anti-bacterial autophagy .
Disease Mechanisms
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Autoimmune Diseases: Antibody-based assays revealed elevated HSPA8 levels in lupus-prone mice, linked to dysregulated CMA and pathogenic autoantibody production .
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Viral Entry: HSPA8 monoclonal antibodies blocked infectious bronchitis virus (IBV) infection by disrupting viral attachment to host cell membranes .
Technical Performance
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Sensitivity: Detects endogenous HSPA8 at concentrations as low as 0.1 ng/µL in Western blotting (WB) .
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Cross-Reactivity: Validated for human, mouse, rat, bovine, and pig samples .
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Functional Assays: Used in co-immunoprecipitation (Co-IP) to map HSPA8 interactions with NLRC4 (inflammasome component) and BECN1 (autophagy protein) .
Limitations and Considerations
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Species Restrictions: While cross-reactive with multiple species, performance in non-mammalian systems remains untested .
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Batch Variability: Commercial products from different vendors show variability in recommended dilutions (e.g., 1:500–1:256K for WB) .
Future Research Directions
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Therapeutic Targeting: Explore HSPA8 inhibition/activation strategies using antibody-guided drug delivery .
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Diagnostic Biomarkers: Investigate HSPA8 overexpression in autoimmune diseases or cancer using IHC-validated antibodies .
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Viral Antagonism: Develop antibody-based inhibitors against HSPA8-dependent viral entry mechanisms .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
This HSPA8 monoclonal antibody was developed using hybridoma technology. The antibody-producing hybridomas were generated by fusing myeloma cells with spleen cells from immunized mice. The mice were immunized with recombinant human HSPA8 protein (amino acids 2-646). This HSPA8 monoclonal antibody is designed to detect endogenous levels of human HSPA8 protein. It has been purified using protein G and rigorously validated for its performance in ELISA, Western blotting, immunohistochemistry, immunofluorescence, and flow cytometry applications. Its purity exceeds 95%.
The HSPA8 protein primarily functions as a chaperone within the cell, assisting in protein folding, transport, and degradation. It accomplishes this by binding to nascent polypeptide chains or misfolded proteins, guiding them through various cellular processes. HSPA8 plays a critical role in maintaining cellular homeostasis by preventing the accumulation of misfolded or aggregated proteins, which can lead to cellular dysfunction and disease. It is also involved in the regulation of apoptosis, antigen processing, and cell signaling.
Target Background
- Data suggest that both the small heat shock protein HspB1/Hsp27 and the constitutive chaperone Hsc70/HspA8 interact with tau to prevent tau-fibril/amyloid formation. Hsc70 is highly efficient at preventing tau-fibril elongation, possibly by capping ends of tau-fibrils. (HspB1 = heat shock protein family B small member 1; Hsc70 = heat shock protein family A Hsp70) PMID: 29298892
- This study demonstrated that particular distribution for HSC70 and PSMC4 in the cytoplasm and accumulation within Lewy body in the dopaminergic neurons of the substantia nigra in Parkinson patients. PMID: 29218503
- Hsp70participated in PINK1-mediated mitophagy by stabilizing PINK1. PMID: 29107085
- These results demonstrate not only an important mechanism of Hsc70 in facilitating EV-A71 replication, but also a target for antiviral drug development. PMID: 29180285
- Post-transcriptional inhibition of HSPA8 expression leads to synaptic vesicle cycling defects in multiple models of amyotrophic lateral sclerosis. PMID: 28978466
- Taken together, these data suggest that the altered hydrogen bonding observed in the Hsc70 C17W mutant (where the connectivity between Mg2+.nucleotide and E175 is also disrupted) could bring about changes to Hsc70 domain communication, affecting peptide association while also limiting ATP hydrolysis. PMID: 29300467
- The roles of the E3 ubiquitin ligase, carboxy-terminus Hsc70 interacting protein (CHIP) in various types of cancers. PMID: 28731191
- Hsc70/Hsp90 chaperones contribute to the conformational and functional maintenance of DeltaF508-CFTR at 37 degrees C. PMID: 28855508
- Engagement of the oligomer by LAP1 triggers ATP hydrolysis and rapid complex disassembly. Thus, the Torsin complex is a highly dynamic assembly whose oligomeric state is tightly controlled by distinctively localized cellular cofactors. PMID: 28814508
- Downregulation of Hsc70, Hsp70, and IL-15 expression at gene and/or protein levels might support the retention of fertilization products in cases of missed abortion and blighted ovum. PMID: 27225940
- STRO-1 binds to immune-precipitated HSC70, and siRNA-mediated knock down of HSPA8 reduced STRO-1 binding. PMID: 28026090
- Synapsin is part of a multiprotein complex enriched in chaperones/cochaperones including Hsc70. Hsc70 chaperone activity is required for the cytosolic slow axonal transport of synapsin. PMID: 28559423
- This study demonstrates a critical role of Hsc70 in SV40 endoplasmic reticulum-to-cytosol penetration and reveals how SGTA controls Hsc70 to impact this process PMID: 28356524
- In fact, DnaJC5 overexpression induced tau release in cells, neurons, and brain tissue, but only when activity of the chaperone Hsc70 was intact and when tau was able to associate with this chaperone. PMID: 27261198
- These results suggest that Bag1 and Bag3 control the stability of the Hsc70-client complex using at least two distinct protein-protein contacts, providing a previously under-appreciated layer of molecular regulation in the human Hsc70 system. PMID: 27474739
- While cerebrospinal fluid Nrf2 and HSPA8 do not appear to offer diagnostic biomarkers for Parkinson's disease (PD), the associations between Nrf2 levels and UPDRS scores in LRRK2 + PD patients merit further investigation PMID: 26526034
- HSPA8 maintains pluripotency of human pluripotent stem cells by binding to the master pluripotency regulator OCT4 and facilitating its DNA-binding activity. PMID: 26549849
- Heat shock cognate 70 (HSC70) is an essential component of Aes foci in colorectal cancer cells. PMID: 26229111
- Cx43-Hsc70 interaction regulates cell cycle G1/S progression through a novel mechanism by which Cx43-Hsc70 interaction prevents the nuclear accumulation of p27 through controlling the nuclear translocation of cyclin D1-CDK4-p27 complex. PMID: 26481195
- Serum HSC71 was identified as a novel serum biomarker of renal cell carcinoma, particularly useful in early diagnosis of clear-cell type. PMID: 26425554
- These findings provide further evidence that histatin 3 may be involved in the regulation of cell proliferation, particularly during G1/S transition, via the ubiquitin-proteasome system of p27(Kip1) and HSC70. PMID: 26775844
- Constitutive heat shock protein HSC70 forms granule-like structures in the cytoplasm of human cells several days after the exposure to heat stress PMID: 26335814
- These results indicate that CHIP decreases the Kv1.5 protein level and functional channel by facilitating its degradation in concert with chaperone Hsc70 PMID: 26232501
- Active Hsc70 requires active tau to regulate microtubule assembly in vivo, suggesting that tau acts in some ways as a co-chaperone for Hsc70 to coordinate microtubule assembly. PMID: 25882706
- Detailed and systematic investigation to characterize if there are significant differences in the CHIP in vitro ubiquitination of human Hsp70 and Hsc70. PMID: 26010904
- HSP70 binds CD91 and TLR4 on decidual CD1a(+) dendritic cells, causes their maturation, and increases IL-15 in the context of Th1 cytokine/chemokine domination, which could support immune response harmful for ongoing pregnancy. PMID: 25737151
- LAP1 mutations are associated with severe dystonia, cerebellar atrophy, and cardiomyopathy. PMID: 26596547
- HSC70 may serve as a molecular switch to modulate endocytic and autophagy pathways, impacting the source of antigens delivered for MHC class II presentation during cellular stress. (Review) PMID: 25953005
- CHIP docks onto Hsp70/Hsc70 and defines a bipartite mode of interaction between TPR domains and their binding partners. PMID: 25684577
- Hsc70 interacted with Rab1A in a chaperone-dependent manner, and Hsc70 knockdown decreased the level of Rab1A and increased the level of its ubiquitination under stress conditions. PMID: 24801886
- Data suggest that inhibition of nestin and heat shock protein HSPA8/HSC71 may be a useful molecular target in the treatment of glioblastomas. PMID: 25527454
- LAP1 and LULL1 regulate Torsin ATPase activity through an active site complementation mechanism. PMID: 25352667
- H-ERG trafficking was impaired by H2O2 after 48 h treatment, accompanied by reciprocal changes of expression between miR-17-5p seed miRNAs and several chaperones (Hsp70, Hsc70, CANX, and Golga20) PMID: 24386440
- The endogenous DNAJC12 and Hsc70 proteins interact in LNCaP cells. PMID: 24122553
- Endoplasmic reticulum-associated degradation of Niemann-Pick C1: evidence for the role of heat shock proteins and identification of lysine residues that accept ubiquitin. PMID: 24891511
- Results identify a systemic hsc70 reduction in Parkinson's disease patients. PMID: 24361989
- Hsp73 sustains Hsp90 chaperone function and critically contributes to cell survival in multiple myeloma patients. PMID: 23065523
- These findings indicate that wild type ILK and the non-oncogenic ILK(R211A) mutation comprise a cardioprotective module with Hsp/c70. PMID: 24260102
- A significant downregulation of HSPA8 and HSPA9 was observed in AD across the three brain regions compared to the controls, suggesting their participation in AD pathogenesis. PMID: 23948933
- HSP8A polymorphism is associated with drug-naive schizophrenia. PMID: 24548631
- Cx43-Hsc70 interaction probably plays a critical role during G1/S progression. PMID: 24056538
- These results demonstrate that Hsp70 recruits PP5 and activates its phosphatase activity, which suggests dual roles for PP5 that might link chaperone systems with signaling pathways in cancer and development. PMID: 24327656
- Altered expression of Hsc70 and eIF5A-1 may cause defects in nucleocytoplasmic transport and play a role in esophageal carcinogenesis. PMID: 23539416
- The U-box mutation stimulated CHIP binding to Hsc70 while promoting CHIP oligomerization. CHIP binding to Hsc70 binding was also stimulated by the presence of an Hsc70 client with a preference for the ADP-bound state. PMID: 23990462
- GLIPR1 interacts with Hsc70, and GLIPR1 overexpression or Hsc70 knockdown leads to transcriptional suppression of AURKA and TPX2. PMID: 23333597
- PKCiota knockdown sensitized cells to oxidative stress-induced apoptosis, whereas forced PKCiota expression counteracted the oxidative stress-induced apoptosis via Hsc70. PMID: 23224638
- Relaxation-based NMR experiments on the Hsc70-CHIP complex determined that the two partners move independently in solution, similar to "beads on a string." PMID: 23865999
- LAP1 and LULL1 as regulatory cofactors that are responsible for the activation of TorA's ATPase activity. PMID: 23569223
- 70-kDa heat shock cognate protein hsc70 mediates calmodulin-dependent nuclear import of the sex-determining factor SRY. PMID: 23235156
- Studies suggest that Hsc70 and lysosome-associated protein 2A (LAMP-2A) through chaperone-mediated autophagy (CMA) play a role in the clearance of Htt and suggest a novel strategy to target the degradation of mutant huntingtin (Htt). PMID: 23071649
