HSP70 Kit ELISA

N° du produit ABIN625462

Détail du produit HSP70 Kit ELISA

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

Reactivité: Souris, Rat, Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 2-600 ng/mL

Seuil minimal de détection: 2 ng/mL

Application: ELISA

Fonction: Human HSP70 ELISA Kit for cell culture supernatants, Heparin and/or EDTA treated plasma, and serum samples. Citrate is not recommended.

Type d'échantillon: Plasma, Cell Culture Supernatant, Serum

Analytical Method: Quantitative

Specificité: This ELISA antibody pair also can recognize mouse and rat HSP70

Sensibilité: 2 ng/mL

Attributs du produit:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Ingrédients:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Matériel non inclus:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Information d'application

Indications d'application: Recommended Dilution for serum and plasma samples2 fold

Volume d'échantillon: 100 μL

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Préparation des réactifs:

1. Bring all reagents and samples to room temperature (18 - 25°C) before use. 2. Sample dilution: If your samples need to be diluted, Assay Diluent C (Item L) is used for dilution of serum/plasma/culture supernatants/urine. 3. Assay Diluent B (Item E) should be diluted 5-fold with deionized or distilled water before use. 4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µl Assay Diluent C (Item L) into Item C vial to prepare a 600 ng/ml standard solution. Dissolve the powder thoroughly by a gentle mix. Pipette 400 µl Assay Diluent C into each tube. Use the 600 ng/ml standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Gently vortex to mix. Assay Diluent C serves as the zero standard (0 ng/ml). 5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer. 6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µl of 1x Assay Diluent B (Item E) into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4°C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure. 7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 25,000-fold with 1x Assay Diluent B (Item E). For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 2 µl of HRP-Streptavidin concentrate into a tube with 198.0 µl 1x Assay Diluent B to prepare a 100-fold diluted HRP-Streptavidin solution (do not store the diluted solution for next day use). Mix through and then pipette 40 µl of prepared 100-fold diluted solution into a tube with 10 ml 1x Assay Diluent B to prepare a final 25,000 fold diluted HRP-Streptavidin solution.

Procédure de l'essai:

1. Bring all reagents and samples to room temperature (18 - 25°C) before use. It is recommended that all standards and samples be run at least in duplicate. 2. Add 100 µl of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4°C with gentle shaking. 3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 µl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels. 4. Add 100 µl of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking. 5. Discard the solution. Repeat the wash as in step 3. 6. Add 100 µl of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking. 7. Discard the solution. Repeat the wash as in step 3. 8. Add 100 µl of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking. 9. Add 50 µl of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Calcul des résultats:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.

Restrictions: For Research Use only

Stockage

Stock: -20 °C

Stockage commentaire: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Date de péremption: 6 months

Références pour HSP70 Kit ELISA (ABIN625462)

Pei, Pan, Zhu, Ding, Liu, Lv, Zou, Luo: "Gemcitabine-treated pancreatic cancer cell medium induces the specific CTL antitumor activity by stimulating the maturation of dendritic cells." dans: International immunopharmacology, Vol. 19, Issue 1, pp. 10-6, (2014) (PubMed).

Détail du antigène

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

Autre désignation: HSP70 (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

Sujet: The Human HSP70 (Heat Shock Protein 70) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human HSP70 in serum, plasma (Collect plasma using EDTA and heparin as an anticoagulant. Citrate are not recommended), cell culture supernatants and urine. This assay employs an antibody specific for human HSP70 coated on a 96-well plate. Standards and samples are pipetted into the wells and HSP70 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human HSP70 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of HSP70 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.

ID gène: 3308

UniProt: P34932