Clusterin Kit ELISA

N° du produit ABIN1979436

Détail du produit Clusterin Kit ELISA

Antigène: Clusterin (CLU)

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 15-20000 pg/mL

Seuil minimal de détection: 15 pg/mL

Application: ELISA

Fonction: Human Clusterin ELISA Kit for cell culture supernatants, plasma, and serum samples.

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

Analytical Method: Quantitative

Specificité: This ELISA antibody pair detect human Clusterin. Other species not determined.

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 samples100,000 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 (Item E) should be used for dilution of serum/plasms/cell culture supernatants/urine. Suggested dilution for normal serum/plasma: 100,000 fold. The Human Clusterin ELISA Kit Protocol 3 For example, add 1 µL of serum/plasma into a tube with 199 µL 1x Assay Diluent to prepare a 200-fold diluted sample. Mix through and then pipette 1 µL of prepared 200-fold diluted sample into a tube with 499 µL 1x Assay Diluent to prepare a final 100,000 fold diluted sample. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator. 3. Assay Diluent (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 1x Assay Diluent (Item E) into Item C vial to prepare a 80 ng/ml standard solution. Dissolve the powder thoroughly by a gentle mix. Add 150 µL Clusterin standard from the vial of Item C, into a tube with 450 µL 1x Assay Diluent to prepare a 20,000 pg/ml standard solution. Pipette 600myl 1x Assay Diluent into each tube. Use the 20,000 pg/ml standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. 1x Assay Diluent serves as the zero standard (0 pg/ml). 200 µL 150 µL standard + 450 µL 200myl 200 µL 200 µL 200 µL 20,000 5,000 1,250 312.5 78.13 19.53 0 pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml The Human Clusterin ELISA Kit Protocol 4 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 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 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 1,000-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 12 µL of HRP-Streptavidin concentrate into a tube with 12 mL 1x Assay Diluent to prepare a 1,000-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.

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 4) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or overnight 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 myl) 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.

Précision du teste: Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: Avoid repeated freeze-thaw cycles.

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 Clusterin Kit ELISA (ABIN1979436)

Oztas, Ozler, Ersoy, Iskender, Sucak, Ergin, Uygur, Danisman: "Increased levels of serum clusterin is associated with intrauterine growth restriction and adverse pregnancy outcomes in preeclampsia." dans: Journal of perinatal medicine, Vol. 44, Issue 3, pp. 269-75, (2016) (PubMed).

Détail du antigène

Antigène: Clusterin (CLU)

Autre désignation: Clusterin (CLU Produits)

Synonymes: LOC100136120 Kit ELISA, clu Kit ELISA, MGC53055 Kit ELISA, zgc:73352 Kit ELISA, xclu Kit ELISA, MGC89163 Kit ELISA, CLU Kit ELISA, APO-J Kit ELISA, APOJ Kit ELISA, CLI Kit ELISA, CLU1 Kit ELISA, CLU2 Kit ELISA, KUB1 Kit ELISA, NA1/NA2 Kit ELISA, SGP-2 Kit ELISA, SGP2 Kit ELISA, SP-40 Kit ELISA, TRPM-2 Kit ELISA, TRPM2 Kit ELISA, DAG Kit ELISA, RATTRPM2B Kit ELISA, SP40 Kit ELISA, TRPM2B Kit ELISA, Trpm2 Kit ELISA, Trpmb Kit ELISA, AI893575 Kit ELISA, ApoJ Kit ELISA, Cli Kit ELISA, D14Ucla3 Kit ELISA, Sgp-2 Kit ELISA, Sgp2 Kit ELISA, Sugp-2 Kit ELISA, GP80 Kit ELISA, gpIII Kit ELISA, 40 Kit ELISA, clusterin-1 Kit ELISA, clusterin L homeolog Kit ELISA, clusterin Kit ELISA, Clusterin Kit ELISA, LOC100136120 Kit ELISA, clu.L Kit ELISA, clu Kit ELISA, CLU Kit ELISA, clus Kit ELISA, Clu Kit ELISA, LOC101113728 Kit ELISA

ID gène: 1191

UniProt: P10909

Pathways: Apoptose, Negative Regulation of intrinsic apoptotic Signaling