HSP70 ELISA Kit

Catalog No. ABIN6973380

Product Details HSP70 ELISA Kit

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

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.156 ng/mL - 10 ng/mL

Minimum Detection Limit: 0.156 ng/mL

Application: ELISA

Purpose: For the quantitative determination of mouse heat shock protein 70 (Hsp-70) concentrations in serum, plasma, tissue homogenates.

Sample Type: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of mouse Hsp-70. No significant cross-reactivity or interference between mouse Hsp-70 and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between mouse Hsp-70 and all the analogues, therefore, cross reaction may still exist.

Sensitivity: 0.039 ng/mL

Components:

• Assay plate
• Standard
• HRP-avidin (100 x concentrate)
• Biotin-antibody (100 x concentrate)
• Sample Diluent
• HRP-avidin Diluent
• Biotin-antibody Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 100 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Reagent Preparation:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. Wash Buffer(1x)- If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 10 ng/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (10 ng/mL). Sample Diluent serves as the zero standard (0 ng/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• 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).

Note:

Recommend to dilute the serum or plasma samples with Sample Diluent (1:100) before test. The suggested 100-fold dilution can be achieved by adding 10 μL sample to 40 μL of Sample Diluent. Complete the 100-fold dilution by adding 15 μL of this solution to 285 μL of Sample Diluent. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments.

Assay Precision: Intra-assay Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date. May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Expiry Date: 6 months

Target Details for HSP70

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

Alternative Name: Heat Shock Protein 70,Hsp-70 (HSP70 Products)

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

Background:

Abbreviation: HSPA1A

Alias: DAQB-147D11.1, FLJ54303, FLJ54370, FLJ54392, FLJ54408, FLJ75127, HSP70-1, HSP70-1A, HSP70I, HSP72, HSPA1, HSPA1B, dnaK-type molecular chaperone HSP70-1|heat shock 70kD protein 1A|heat shock-induced

UniProt: Q61696