HSP70 Kit ELISA

N° du produit ABIN7067012

Détail du produit HSP70 Kit ELISA

Antigène: HSP70 (Heat Shock Protein 70 (HSP70))

Reactivité: Boeuf (Vache), Chévre, Mouton

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 3.12 ng/mL - 200 ng/mL

Seuil minimal de détection: 3.12 ng/mL

Application: ELISA

Fonction: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of HSP70 in serum, plasma, tissue homogenates, cell lysates, cell culture supernates. Due to the 98% homology of the sequence among different species, the kit can be used to detect bovine, caprine and ovine samples.

Type d'échantillon: Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificité: This assay has high sensitivity and excellent specificity for detection of Heat Shock Protein 70 (HSP70)

Sensibilité: 1.31 ng/mL

Ingrédients:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Information d'application

Commentaires:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Volume d'échantillon: 100 μL

Durée du test: 3 h

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Préparation des réactifs:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 200 ng/mL. Prepare 7 tubes containing 0.5 mL Standard Diluent and produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 200 ng/mL, 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.12 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Préparation de l'échantillon:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Précision du teste: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Restrictions: For Research Use only

Stockage

Précaution d'utilisation: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Stock: 4 °C/-20 °C

Stockage commentaire:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Date de péremption: 6 months

Références pour HSP70 Kit ELISA (ABIN7067012)

Gonzalez-Rivas, DiGiacomo, Giraldo, Leury, Cottrell, Dunshea: "Reducing rumen starch fermentation of wheat with three percent sodium hydroxide has the potential to ameliorate the effect of heat stress in grain-fed wethers." dans: Journal of animal science, Vol. 95, Issue 12, pp. 5547-5562, (2018) (PubMed).

Lamy, Jurkovich, Rodrigues, Geraldo, Cachucho, Silva, Matos, Capela E Silva, Pinheiro, Könyves, Bakony, Pereira: "Detection of 70 kDa heat shock protein in the saliva of dairy cows." dans: The Journal of dairy research, Vol. 84, Issue 3, pp. 280-282, (2018) (PubMed).

Hu, Zhang, Zheng, Cheng, Wang: "The effect of heat stress on gene expression and synthesis of heat-shock and milk proteins in bovine mammary epithelial cells." dans: Animal science journal = Nihon chikusan Gakkaihō, Vol. 87, Issue 1, pp. 84-91, (2016) (PubMed).

Horst, Eschbach, Pfeifer, Hübenthal, Sassen, Steinfeldt, Wulf, Ruchholtz, Pape, Hildebrand: "Local inflammation in fracture hematoma: results from a combined trauma model in pigs." dans: Mediators of inflammation, Vol. 2015, pp. 126060, (2015) (PubMed).

Détail du antigène

Antigène: HSP70 (Heat Shock Protein 70 (HSP70))

Abstract: HSP70 Produits

Synonymes: APG-2 Kit ELISA, HS24/P52 Kit ELISA, HSPH2 Kit ELISA, RY Kit ELISA, hsp70 Kit ELISA, hsp70RY Kit ELISA, CG31354 Kit ELISA, HSP70 Kit ELISA, Hsp70Bb Kit ELISA, hsp70B Kit ELISA, hsp70Bb-prime Kit ELISA, DmelCG5834 Kit ELISA, CG5834 Kit ELISA, HSPA1 Kit ELISA, HSP70B' Kit ELISA, HSPA6 Kit ELISA, ARABIDOPSIS HEAT SHOCK PROTEIN 70 Kit ELISA, ATHSP70 Kit ELISA, heat shock protein 70 Kit ELISA, LOC100305036 Kit ELISA, hsc70 Kit ELISA, Hsp70 Kit ELISA, Hsp70-1 Kit ELISA, Hsp70.1 Kit ELISA, hsp68 Kit ELISA, Hsp110 Kit ELISA, irp94 Kit ELISA, HSP70-2 Kit ELISA, HSPA1B Kit ELISA, HSPA2 Kit ELISA, hsp70-5 Kit ELISA, HSP70-1 Kit ELISA, HSP70.1 Kit ELISA, HSP70.2 Kit ELISA, heat shock protein family A (Hsp70) member 4 Kit ELISA, CG5834 gene product from transcript CG5834-RA Kit ELISA, heat shock protein 70 Kit ELISA, heat shock protein family A (Hsp70) member 6 Kit ELISA, heat shock 70kDa protein 2 Kit ELISA, heat shock 70 kD protein cognate Kit ELISA, Hsp70 family chaperone Kit ELISA, Heat shock protein 70 Kit ELISA, Heat shock protein 70, putative Kit ELISA, heat shock protein 1B Kit ELISA, heat shock protein family A member 4 Kit ELISA, heat shock 70kDa protein 1A Kit ELISA, heat shock protein 1 Kit ELISA, Heat Shock Protein Kit ELISA, heat shock cognate 70-kd protein Kit ELISA, Heat shock 70 kDa protein 1A Kit ELISA, HSPA4 Kit ELISA, Hsp70Bbb Kit ELISA, HSP70 Kit ELISA, HSPA6 Kit ELISA, HSPA2 Kit ELISA, PCC7424_2419 Kit ELISA, Isop_1041 Kit ELISA, CGB_C3390W Kit ELISA, Bacsa_1698 Kit ELISA, dnaK-B Kit ELISA, LOC100305036 Kit ELISA, Hspa1b Kit ELISA, Hspa4 Kit ELISA, HSPA1A Kit ELISA, hsp1 Kit ELISA, hsp-70 Kit ELISA, hsp70 Kit ELISA, LOC108348108 Kit ELISA