PODXL Kit ELISA

N° du produit ABIN956203

Détail du produit PODXL Kit ELISA

Antigène: PODXL (Podocalyxin-Like (PODXL))

Reactivité: Rat

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Fonction: This ELISA kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of rat PCX in biological samples.

Analytical Method: Quantitative

Attributs du produit: The assay sample and buffer are incubated together in the anti-PCX antibody coated plate for sixty minutes and washed. The diluted C-Peptide-HRP conjugate is then added to each well and incubated. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for the enzyme. The product of the enzyme-substrate reaction is a blue colored complex. Finally, a stopping solution is added to stop the reaction. The stop solution changes the color from blue to yellow. The intensity of the color is then measured spectrophotometrically at 450 nm in a microplate reader. The intensity of the color is inversely proportional to the PCX concentration since PCX from calibrators and samples compete with the PCX-HRP conjugate for the anti-PCX antibody binding site. Since the number of sites is limited, as more sites are occupied by PCX from the samples or calibrators, fewer sites are available to bind the C-Peptide-HRP conjugate. Calibrators of known PCX concentrations are run concurrently with the samples being assayed and a calibration curve is plotted relating the intensity of the color (OD) to the concentration of C-Peptide. The unknown PCX concentration in each sample is interpolated from this curve.

Ingrédients: Pre-coated 96-well plate 1 Calibrator 1 (0 ng/mL)
Calibrator 2 (5 ng/mL)
Calibrator 3 (10 ng/mL)
Calibrator 4 (25 ng/mL)
Calibrator 5 (50 ng/mL)
Calibrator 6 (100 ng/mL)
Enzyme Conjugate (1 x 6 mL)
Substrate A (1 x 6 mL)
Substrate B (1 x 6 mL)
Stop Solution (1 x 6 mL)
Wash Buffer (100X concentrate) (1 x 6 mL)
Lysis Buffer 1 x 10 mL.

Matériel non inclus: 1. Microplate reader with 450 nm filter.
2. Precision pipettes to deliver 1-2 mL volumes.
3. Adjustable 10-100 mL pipettes for reagent preparation.
4. 100 mL and 1 L graduated cylinders.
5. Calibrated adjustable precision pipettes with disposable tips (multi-channel is desirable for large assays). 6. 37°C incubator.
7. Absorbent paper.
8. Distilled or deionized water
9. Data analysis tools such as graphing software, or graph paper (linear, log-log, semi-log, or log-logit as desired).
10. Tubes to prepare Calibrators or sample dilutions.

Information d'application

Plaque: Pre-coated

Préparation des réactifs:

All reagents must be allowed to reach room temperature before use. Additional information for individual reagents can be found on vial labels. Note: 1. This kit contains materials with small quantities of sodium azide. Sodium azide reacts with lead and copper plumbing to form explosive metal azides. Upon disposal, flush drain with a large volume of water to prevent azide accumulation. Avoid ingestion and contact with eyes, skin, and mucous membranes. In case of contact, rinse affected area with plenty of water. Observe all federal, state, and local regulations for disposal.
2. All blood components and biological materials should be handled as potentially hazardous. Follow universal precautions as established by the Centers for Disease Control and Prevention and by the Occupational Safety and Health Administration when handling and disposing of infectious agents.

Procédure de l'essai:

1. Prepare all calibrators before starting assay procedure (see Reagent Preparation). It is recommended that all calibrators and samples be added in duplicate to the microtiter plate.
   2. First, secure the desired number of coated wells in the holder, then add 100 µL of calibrators and samples to the appropriate well of the antibody coated microtiter plate.
   3. Add 50 µL of conjugate to each well. Mix well. Complete mixing in this step is important. Cover and incubate for 1 hour at 37°C.
   4. Prepare substrate solution no more than 15 minutes before end of incubation (see Reagent Preparation).
   5. Wash the microtiter plate using one of the specified methods indicated below:
   6. Manual Washing: Remove incubation mixture by aspiration contents of the plate into a sink or proper waste container. Using a squirt bottle, fill each well completely with distilled or de-ionized water, then aspirate contents of the plate into a sink or proper waste container. Repeat this procedure four more times for a total of five washes. After final wash, invert plate, and blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears. Note: Hold the sides of the plate frame firmly when washing the plate to assure that all strips remain securely in frame.
   7. Automated Washing: Aspirate all wells, then wash plate five times using distilled or de-ionized water. Always adjust your washer to aspirate as much liquid as possible and set fill volume to 350 µL/well/wash (range: 350-400 µL). After final wash, invert plate, and blot dry by hitting plate onto absorbent paper and paper towels until no moisture appears. It is recommended that the washer be set for a soaking time of 15-30° Conds or shaking time of 5 seconds between washes.
   8. Add 50 µL substrate A and B to each well. Cover and incubate for 15 minutes at 20-25°C.
   9. Add 50 µL stop solution to each well. Mix well.
   10. Read the optical density (OD) at 450 nm using a microtiter plate reader within 30 minutes.

Calcul des résultats:

1. Calculate the mean absorbance value A450 for each set of calibrators and samples.
   2. Divide the average A450 value for each calibrator, control and sample by the average A450 of calibrator 0 and multiply by 100 to obtain %B/B0 for each sample.
   3. Prepare a calibration curve by plotting the average absorbance of each calibrator versus the corresponding concentrations of the calibrators on linear-log graph paper or the %B/B0 value for each calibrator versus the corresponding concentration of the calibrator on linear-log or logit-log paper. logit= ln(B/B0)/(1-B/B0).
   4. Any values obtained for diluted samples must be further converted by applying the appropriate dilution factor in the calculations.
   5. The calibrator density is on the x-axis, while the B/B0 is on the y-axis
   6. The sensitivity of this assay is 0.01 ng/mL

Restrictions: For Research Use only

Stockage

Stock: 4 °C

Stockage commentaire: All reagents should be stored at 4°C upon receipt. For expiration date refer to kit label.

Date de péremption: The expiry date is stated on the label.

Détail du antigène

Antigène: PODXL (Podocalyxin-Like (PODXL))

Autre désignation: Podocalyxin (PODXL Produits)

Synonymes: Gp200 Kit ELISA, PC Kit ELISA, PCLP Kit ELISA, PCLP-1 Kit ELISA, PCB Kit ELISA, AW121214 Kit ELISA, Ly102 Kit ELISA, Pclp1 Kit ELISA, podocalyxin Kit ELISA, MEP21 Kit ELISA, PODXL Kit ELISA, Gp135 Kit ELISA, PCLP1 Kit ELISA, cPCLP1 Kit ELISA, podxl Kit ELISA, fc18d02 Kit ELISA, wu:fc18d02 Kit ELISA, Pc Kit ELISA, Pcb Kit ELISA, Podxl Kit ELISA, podocalyxin like Kit ELISA, pyruvate carboxylase Kit ELISA, podocalyxin-like Kit ELISA, PODXL Kit ELISA, PC Kit ELISA, Podxl Kit ELISA, podxl Kit ELISA, Pcx Kit ELISA

Pathways: Tube Formation