Vitamin D-Binding Protein Kit ELISA

N° du produit ABIN5564700

Détail du produit Vitamin D-Binding Protein Kit ELISA

Antigène: Vitamin D-Binding Protein (GC)

Reactivité: Cheval

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 6.25 ng/mL - 400 ng/mL

Seuil minimal de détection: 6.25 ng/mL

Application: ELISA

Type d'échantillon: Plasma, Serum

Analytical Method: Quantitative

Specificité:

This assay has high sensitivity and excellent specificity for detection of Vitamin D Binding Protein (DBP).
No significant cross-reactivity or interference between Vitamin D Binding Protein (DBP) and analogues was observed.

Sensibilité: 2.84 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

Durée du test: 3 h

Plaque: Pre-coated

Protocole: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Vitamin D Binding Protein (DBP). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Vitamin D Binding Protein (DBP). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Vitamin D Binding Protein (DBP), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Vitamin D Binding Protein (DBP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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 400 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 400 ng/mL, 200 ng/mL, 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 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.

Précision du teste:

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vitamin D Binding Protein (DBP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Vitamin D Binding Protein (DBP) 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.

Conseil sur la manipulation: The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Stock: 4 °C

Stockage commentaire:

• For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
• For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
  Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
• For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.

Date de péremption: 6 months

Détail du antigène

Antigène: Vitamin D-Binding Protein (GC)

Autre désignation: Vitamin D Binding Protein (DBP) (GC Produits)

Synonymes: DBP Kit ELISA, DBP/GC Kit ELISA, GRD3 Kit ELISA, VDBG Kit ELISA, VDBP Kit ELISA, zgc:110389 Kit ELISA, zgc:92753 Kit ELISA, GC Kit ELISA, Gc Kit ELISA, DBP02 Kit ELISA, VDB Kit ELISA, Vdbp Kit ELISA, VTDB Kit ELISA, gc Kit ELISA, GC, vitamin D binding protein Kit ELISA, D-box binding PAR bZIP transcription factor Kit ELISA, D site albumin promoter binding protein Kit ELISA, group-specific component (vitamin D binding protein) Kit ELISA, group specific component Kit ELISA, group-specific component (vitamin D binding protein) S homeolog Kit ELISA, GC Kit ELISA, Dbp Kit ELISA, gc Kit ELISA, Gc Kit ELISA, gc.S Kit ELISA

Pathways: Metabolism of Steroid Hormones and Vitamin D, Monocarboxylic Acid Catabolic Process