Tel:
+49 (0)241 95 163 153
Fax:
+49 (0)241 95 163 155
E-Mail:
orders@anticorps-enligne.fr

PODXL Kit ELISA

PODXL Reactivité: Humain Colorimetric Sandwich ELISA 0.156-10 ng/mL Urine
N° du produit ABIN456213
  • Antigène Voir toutes PODXL Kits ELISA
    PODXL (Podocalyxin-Like (PODXL))
    Reactivité
    • 5
    • 4
    • 4
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    0.156-10 ng/mL
    Seuil minimal de détection
    0.156 ng/mL
    Application
    ELISA
    Fonction
    This immunoassay kit allows for the specific measurement of human Podocalyxin concentrations in urine.
    Type d'échantillon
    Urine
    Analytical Method
    Quantitative
    Specificité
    This assay recognizes recombinant and natural human Podocalyxin.
    Réactivité croisée (Details)
    No significant cross-reactivity or interference was observed.
    Sensibilité
    < 78 pg/mL
    The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest detectable concentration that could be differentiated from zero.
    Attributs du produit
    Homo sapiens,Human,Podocalyxin,GCTM-2 antigen,Gp200,Podocalyxin-like protein 1,PC,PCLP-1,PODXL,PCLP,PCLP1
    Ingrédients
    Reagent (Quantity): Assay plate (1), Standard (2), Sample Diluent (1x20ml), Assay Diluent A (1x10ml), Assay Diluent B (1x10ml), Detection Reagent A 1×120µl Detection Reagent B 1×120µl Wash Buffer(25 x concentrate) (1x30ml), Substrate (1x10ml), 2 Stop Solution (1x10ml)
    Featured
    Discover our best selling PODXL Kit ELISA
    Top Product
    Discover our top product PODXL Kit ELISA
  • Volume d'échantillon
    100 μL
    Plaque
    Pre-coated
    Protocole
    This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for Podocalyxin has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Podocalyxin present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for Podocalyxin is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of Podocalyxin bound in the initial step. The color development is stopped and the intensity of the color is measured.
    Préparation des réactifs

    Bring all reagents to room temperature before use. Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 500 mL of Wash Buffer. Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution of 10,000 pg/mL. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions. The undiluted standard serves as the high standard (10,000 pg/mL). The Sample Diluent serves as the zero standard (0 pg/mL). Detection Reagent A and B - Dilute to the working concentration specified on the vial label using Assay Diluent A and B (1:100), respectively.

    Prélèvement de l'échantillon
    Urine - Aseptically collect the first urine of the day (mid-stream), voided directly into a sterile container. Centrifuge to remove particulate matter, assay immediately or aliquot and store at ≤ -20 °C. Avoid repeated freeze-thaw cycles.
    Procédure de l'essai

    Allow all reagents to reach room temperature. Arrange and label required number of strips.
    1. Prepare all reagents, working standards and samples as directed in the previous sections.
    2. Add 100 uL of Standard, Control, or sample per well. Cover with the adhesive strip. Incubate for 2 hours at 37 °C.
    3. Remove the liquid of each well, don’t wash.
    4. Add 100 uL of Detection Reagent A to each well. Incubate for 1 hour at 37°C. Detection Reagent A may appear cloudy. Warm to room temperature and mix gently until solution appears uniform.
    5. Aspirate each well and wash, repeating the process three times for a total of three washes. Wash by filling each well with Wash Buffer (350 uL) using a squirt bottle, multi-channel pipette, manifold dispenser 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. 3 Invert the plate and blot it against clean paper towels.
    6. Add 100 uL of Detection Reagent B to each well. Cover with a new adhesive strip.Incubate for 1 hours at 37 °C.
    7. Repeat the aspiration/wash as in step
    5. 8. Add 90 uL of Substrate Solution to each well. Incubate for 30 minutes at room temperature. Protect from light.
    9. Add 50 uL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
    10. Determine the optical density of each well within 30 minutes, using a microplate reader set to 450 nm.
    Important Note:
    1. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
    2. It is recommended that no more than 32 wells be used for each assay run if manual pipetting is used since pipetting of all standards, specimens and controls should be completed within 5 minutes. A full plate of 96 wells may be used if automated pipetting is available.
    3. Duplication of all standards and specimens, although not required, is recommended.
    4. When mixing or reconstituting protein solutions, always avoid foaming.
    5. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent.
    6. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary.

    Calcul des résultats

    Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, 4 construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the Podocalyxin concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

    Restrictions
    For Research Use only
  • Conseil sur la manipulation
    1. The kit should not be used beyond the expiration date on the kit label.
    2. Do not mix or substitute reagents with those from other lots or sources.
    3. If samples generate values higher than the highest standard, further dilute the samples with the Assay Diluent and repeat the assay. Any variation in standard diluent, operator, pipetting technique, washing technique,incubation time or temperature, and kit age can cause variation in binding.
    4. This assay is designed to eliminate interference by soluble receptors, ligands, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.
    Stock
    4 °C/-20 °C
    Stockage commentaire
    The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 °C upon being received. The other reagents can be stored at 4 °C.
  • Antigène Voir toutes PODXL Kits ELISA
    PODXL (Podocalyxin-Like (PODXL))
    Autre désignation
    PODXL (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
    Sujet
    Podocalyxin, also known as Podocalyxin-like protein-1 (PCLP1 or PODXL), is a type I transmembrane glycoprotein. It belongs to the CD34/Podocalyxin family of sialomucins that share structural similarity and sequence homology. Podocalyxin is a major sialoprotein in the podocytes of the kidney glomerulus and is also expressed by both endothelium and multipotent hematopoietic progenitors. It has been identified as a novel cell surface marker for hemangioblasts, the common precursors of hematopoietic and endothelial cells.
    Pathways
    Tube Formation
Vous êtes ici:
Support technique