TTR Kit ELISA

N° du produit ABIN7041519

Détail du produit TTR Kit ELISA

Antigène: TTR (Transthyretin (TTR))

Reactivité: Rat

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 15.625 ng/mL - 500 ng/mL

Seuil minimal de détection: 15.625 ng/mL

Application: ELISA

Fonction: The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of prealbumin in rat serum, plasma.

Type d'échantillon: Plasma, Serum, Urine

Analytical Method: Quantitative

Ingrédients:

• ELISA Micro Plate antibody coated
• Enzyme Conjugated Detection Antibody (100X)
• Calibrator
• Diluent Concentrate (5X)
• Wash Solution Concentrate (20X)
• Chromogen - Substrate Solution
• STOP Solution

Matériel non inclus:

1. Microplate reader with 450 ± 10nm filter.
2. Precision single or multi-channel pipettes and disposable tips.
3. Microcentrifuge tubes for diluting samples.
4. Squirt bottle or Microtitre washer
5. Deionized or distilled water.
6. Container for Wash Solution
7. Centrifuge for sample collection
8. Anticoagulant for plasma collection
9. Incubator capable of maintaining 37 °C.
10. Microplate shaker

Information d'application

Volume d'échantillon: 100 μL

Durée du test: 1 h

Plaque: Pre-coated

Protocole: In this assay the prealbumin (PRE) present in samples reacts with the anti-PRE antibodies, which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-PRE antibodies conjugated with horseradish peroxidase (HRP) are added. These enzyme-labeled antibodies form complexes with the previously bound PRE. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromogenic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of PRE in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of PRE in the test sample. The quantity of PRE in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.

Préparation des réactifs:

• Bring all reagents to room temperature (16°C to 25°C) before use.
• Diluent Concentrate - The Diluent Solution supplied is a 5X Concentrate and must be diluted 1/5 with distilled or deionized water (1 part buffer concentrate, 4 parts dH2O).
• Wash Solution Concentrate - The Wash Solution supplied is a 20X Concentrate and must be diluted 1/20 with distilled or deionized water (1 part buffer concentrate, 19 parts dH2O). Crystal formation in the concentrate may occur when storage temperatures are low. Warming of the concentrate to 30-35°Cbefore dilution can dissolve crystals.
• Enzyme-Antibody Conjugate - Calculate the required amount of working conjugate solution for each microtitre plate test strip by adding 10 µL Enzyme-Antibody Conjugate to 990 µL of 1X Diluent for each test strip to be used for testing. Dilute immediately before use and protect from light. Mix uniformly, but gently. Avoid foaming.
• Pre-coated ELISA Micro Plate - Ready to use as supplied. Unseal foil pouch and remove plate from pouch. Remove all strips and wells that will not be used in the assay and place back in pouch and re-seal along with desiccant.
• Calibrator – Prepare according to the lot specific Certificate of Analysis.

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).

Note:

The assay requires that each test sample be diluted before use. All samples should be assayed in duplicate each time the assay is performed. The recommended dilutions are only suggestions. Dilutions should be based on the expected concentration of the unknown sample such that the diluted sample falls within the dynamic range of the standard curve. If unsure of sample level, a serial dilution with one or two representative samples before running the entire plate is highly recommended.
Serum samples – Recommended starting dilution is 1/5,000. To prepare a 1/5,000 dilution of a sample, transfer 5 µL of sample to 495 µL of 1X diluent. This gives you a 1/100 dilution. Next, dilute the 1/100 by transferring 10 µL into 490 µL of 1X diluent. This gives you a 1/5,000 dilution. Mix thoroughly each stage.
Plasma samples – Recommended starting dilution is 1/5,000. To prepare a 1/5,000 dilution of a sample, transfer 5 µL of sample to 495 µL of 1X diluent. This gives you a 1/100 dilution. Next, dilute the 1/100 by transferring 10 µL into 490 µL of 1X diluent. This gives you a 1/5,000 dilution. Mix thoroughly each stage.

Procédure de l'essai:

1. All samples and standards should be assayed in duplicates.
   2. The Standards and the test sample(s) should be loaded into the ELISA wells as quickly as possible to avoid a shift in OD readings. Using a multichannel pipette would reduce this occurrence. Pipette 100 μL of Standard 0 (0.0 ng/mL) in duplicate Standard 1 (15.63 ng/mL) in duplicate Standard 2 (31.25 ng/mL) in duplicate Standard 3 (62.50 ng/mL) in duplicate Standard 4 (125 ng/mL) in duplicate Standard 5 (250 ng/mL) in duplicate Standard 6 (500 ng/mL) in duplicate
   3. Pipette 100 μL of sample (in duplicate) into pre designated wells.
   4. Incubate the micro titer plate at room temperature for thirty (30 ± 2) minutes. Keep plate covered and level during incubation.
   5. Following incubation, aspirate the contents of the wells.
   6. Completely fill each well with appropriately diluted Wash Solution and aspirate. Repeat three times, for a total of four washes. If washing manually: completely fill wells with wash buffer, invert the plate then pour/shake out the contents in a waste container. Follow this by sharply striking the wells on absorbent paper to remove residual buffer. Repeat 3 times for a total of four washes.
   7. Pipette 100 μL of appropriately diluted Enzyme-Antibody Conjugate to each well. Incubate at room temperature for twenty (20 ± 2) minutes. Keep plate covered in the dark and level during incubation.
   8. Wash and blot the wells as described in Steps 5/6.
   9. Pipette 100 μL of TMB Substrate Solution into each well.
   10. Incubate in the dark at room temperature for precisely ten (10) minutes.
   11. After ten minutes, add 100 μL of Stop Solution to each well.
   12. Determine the absorbance (450 nm) of the contents of each well within 30 minutes. Calibrate the plate reader to manufacturer’s specifications.

Restrictions: For Research Use only

Stockage

Stock: 4 °C

Stockage commentaire:

1. For unopened kit: All reagents should be stored according to the labels on the vials. All reagents should be stored at 4 °C for long term storage. Keep away from heat or direct sunlight.
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

Détail du antigène

Antigène: TTR (Transthyretin (TTR))

Autre désignation: Prealbumin (TTR Produits)

Synonymes: zgc:103583 Kit ELISA, TTR Kit ELISA, LOC100226992 Kit ELISA, CTS Kit ELISA, CTS1 Kit ELISA, HsT2651 Kit ELISA, PALB Kit ELISA, TBPA Kit ELISA, AA408768 Kit ELISA, AI787086 Kit ELISA, D17860 Kit ELISA, prealbumin Kit ELISA, Lr1 Kit ELISA, Tbpa Kit ELISA, Prealbumin Kit ELISA, xTTR Kit ELISA, transthyretin (prealbumin, amyloidosis type I) Kit ELISA, transthyretin Kit ELISA, Transthyretin Kit ELISA, transthyretin (prealbumin, amyloidosis type 1) Kit ELISA, transthyretin-like Kit ELISA, transthyretin L homeolog Kit ELISA, ttr Kit ELISA, TTR Kit ELISA, vpb38 Kit ELISA, MRAD2831_RS38185 Kit ELISA, Caci_4246 Kit ELISA, LOC100226992 Kit ELISA, Ttr Kit ELISA, ttr.L Kit ELISA

Pathways: Hormone Transport