Introduction
Boster's ELISA Kit is for the detection of cytoplasmic Plant Hsp70 from plant extracts. But this kit is also compatible with human tissue extracts and lysates and serum and plasma. Each kit contains sufficient components to quantitate the Hsp70 concentration in up to 40 samples, tested in duplicate.
Overview
| Product Name |
HSP70 ELISA Kit (PLANT)
See all primary antibodies, ELISA kits and proteins |
| SKU/Catalog Number |
EK7115 |
| Storage & Handling |
Store at 4°C. |
| Description |
Colorimetric detection used to quantitate HSP70 in plants. 96wells/kit, with removable strips. |
| Cite This Product |
HSP70 ELISA Kit (PLANT) (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7115)
|
Product Specs
| Reactivity/Species |
Alfalfa, Flax, Human, Maize, Plant |
| Applications |
ELISA
*Our Boster Guarantee covers the use of this product in the above
tested applications.
**For positive and negative control design, consult "Tissue specificity" under
"Protein Target Info" tab. |
| Sample Type |
Cell lysates, Plant extract, Plasma, Serum, Tissue |
| Sensitivity |
0.18 ng/ml |
| Assay Range |
1.563 - 100 ng/ml |
| Cross Reactivity |
There is no detectable cross-reactivity. |
| Pack Size |
96wells/kit |
Gene/Protein Basic Information For Hspa1b (Source: Uniprot.org, NCBI)
| Uniprot Id | P0DMV8/P0DMV9 |
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| Gene Name | Hspa1b |
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| Subcellular Localization | Cytoplasm. |
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*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".
Ontology For Hspa1b (Source: Uniprot.org, NCBI)
| Protein 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, activation of proteolysis of misfolded proteins and the formation and dissociation of protein complexes. 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. This is achieved through cycles of ATP binding, ATP hydrolysis and ADP release, mediated by co-chaperones. The co-chaperones have been shown to not only regulate different steps of the ATPase cycle, but they also have an individual specificity such that one co-chaperone may promote folding of a substrate while another may promote degradation. The affinity 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. It goes through repeated cycles of ATP hydrolysis and nucleotide exchange, which permits cycles of substrate binding and release. The co-chaperones are of three types: J-domain co-chaperones such as 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:24012426, PubMed:26865365, PubMed:24318877). Maintains protein homeostasis during cellular stress through two opposing mechanisms: protein refolding and degradation. Its acetylation/deacetylation state determines whether it functions in protein refolding or protein degradation by controlling the competitive binding of co-chaperones HOPX and STUB1. During the early stress response, the acetylated form binds to HOPX which assists in chaperone-mediated protein refolding, thereafter, it is deacetylated and binds to ubiquitin ligase STUB1 that promotes ubiquitin-mediated protein degradation (PubMed:27708256). Regulates centrosome integrity during mitosis, and is required for the maintenance of a functional mitotic centrosome that supports the assembly of a bipolar mitotic spindle (PubMed:27137183). Enhances STUB1-mediated SMAD3 ubiquitination and degradation and facilitates STUB1-mediated inhibition of TGF-beta signaling (PubMed:24613385). Essential for STUB1-mediated ubiquitination and degradation of FOXP3 in regulatory T-cells (Treg) during inflammation (PubMed:23973223). Negatively regulates heat shock- induced HSF1 transcriptional activity during the attenuation and recovery phase period of the heat shock response (PubMed:9499401). |
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| Tissue Specificity | HSPA1B is testis-specific. |
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| Background | HSP70 genes encode abundant heat-inducible 70-kDa HSPs (HSP70s). In most eukaryotes HSP70 genes exist as part of a multigene family. They are found in most cellular compartments of eukaryotes including nuclei, mitochondria, chloroplasts, the endoplasmic reticulum and the cytosol, as well as in bacteria. The genes show a high degree of conservation, having at least 5O% identity. The N-terminal two thirds of HSP70s are more conserved than the C-terminal third. HSP70 binds ATP with high affinity and possesses a weak ATPase activity which can be stimulated by binding to unfolded proteins and synthetic peptides. When HSC70 (constitutively expressed) present in mammalian cells was truncated, ATP binding activity was found to reside in an N-terminal fragment of 44kDa which lacked peptide binding capacity. Polypeptide binding ability therefore resided within the C-terminal half. The structure of this ATP binding domain displays multiple features of nucleotide binding proteins. All HSP70s, regardless of location, bind proteins, particularly unfolded ones. The molecular chaperones of the HSP70 family recognize and bind to nascent polypeptide chains as well as partially folded intermediates of proteins preventing their aggregation and misfolding. The binding of ATP triggers a critical conformational change leading to the release of the bound substrate protein. The universal ability of HSP70s to undergo cycles of binding to and release from hydrophobic stretches of partially unfolded proteins determines their role in a great variety of vital intracellular functions such as protein synthesis, protein folding and oligomerization and protein transport. |
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Kit Components And Assay QC Data Details Of HSP70 ELISA Kit (PLANT)
*The quality control (QC) data in this section is obtained from Boster's internal QC results and is for reference only. It may differ from the users' lab test results. The users can expect to generate data with similar linearity and quality demonstrated in the typical data but may not achieve exactly the same O.D. values.
| Description | Quantity |
| Anti-Hsp70 Immunoassay Plate | 1 Plate |
| 5X Hsp70 Extraction Reagent | 1 vial/10 ml |
| Recombinant Hsp70 (Alfalfa) Standard | 2 vials |
| Standard and Sample Diluent | 1 vial/ 50 ml |
| 10X Wash Buffer Concentrate | 1 vial/100 ml |
| Anti-Hsp70 Biotinylated Antibody Concentrate | 1 vial/150 µl |
| Anti-Hsp70 Biotinylated Antibody Diluent | 1 vial/ 13 ml |
| Streptavidin: HRP Concentrate | 1 vial/150 µl |
| Streptavidin: HRP Diluent | 1 vial/ 13 ml |
| TMB Substrate | 1 vial/ 13 ml |
| Stop Solution | 1 vial/ 13 ml |
- Microplate reader in standard size.
- Automated plate washer.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- Washing buffer (neutral PBS or TBS).
- Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
- Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.
This data is generated from a recent batch of EK7115 HSP70 ELISA Kit (PLANT). It is not necessarily the same data set used to generate the standard curve shown in the image of this product. TMB reaction incubate at 37°C for 30 minutes
We measured random samples of HSP70 ELISA Kit (PLANT) within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
We measured the performance consistency of HSP70 ELISA Kit (PLANT) to ensure the reproducibility of the assays. Three samples with differing target protein concentrations were assayed using four different lots.
Specific Protocols
Boster provides comprehensive technical information for WB, IHC/IF/ICC, Flow Cytometry sample preparation protocols, assay protocols, troubleshooting tips and assay optimization tips.
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