PF4 Kit ELISA

N° du produit ABIN1979825

Détail du produit PF4 Kit ELISA

Antigène: PF4 (Platelet Factor 4 (PF4))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 20-15000 pg/mL

Seuil minimal de détection: 20 pg/mL

Application: ELISA

Fonction: Human Platelet Factor 4 (CXCL4) ELISA Kit for cell culture supernatants, plasma, and serum samples.

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

Analytical Method: Quantitative

Specificité: This ELISA kit shows no cross-reactivity with any of the following cytokines tested: human Angiogenin, BDNF, BLC, ENA-78, FGF- 4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, MMP-1, - 2, -3, -10, PARC, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Sensibilité: < 20 pg/mL

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 samples200 - 2,000 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 C (Item L) should be used for dilution of serum/plasma/culture supernatants/urine. Suggested dilution for normal serum/plasma: 200-2000 fold. 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 B (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 Assay Diluent C (Item L) into Item C vial to prepare a 140 ng/mL standard solution. Dissolve the powder thoroughly by a gentle mix. Add 75 µL PF-4 standard from the vial of Item C, into a tube with 625 µL Assay Diluent C to prepare a 15,000 pg/mL standard solution. Pipette 400myl Assay Diluent C into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent C serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 75 µL standard + 625 µL 15,000 5,000 1667 555.6 185.2 61.73 20.58 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
   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 B (Item E) 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 B 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 500-fold with 1x Assay Diluent B (Item E). For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 20 µL of HRP-Streptavidin concentrate into a tube with 10 ml 1x Assay Diluent B to prepare a 500-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 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night 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
   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
   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.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent C Human PF-4 concentration (pg/mL) 10 100 1000 10000 O D =4 50 n m 0.001 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of PF-4 is typically less than 20 pg/mL.
Recovery: Recovery was determined by spiking various levels of PF-4 into normal human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 122.4 110-130 Plasma 119.6 111-127 Cell culture media 108.2 101-116
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 121.5 125.2 130.2 Range ( %) 112-30 114-132 116-138 1:4 Average % of Expected 77.28 82.84 86.55 Range ( %) 70-89 74-91 78-95
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

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 PF4 Kit ELISA (ABIN1979825)

Spaks, Jaunalksne, Spaka, Chudasama, Pirtnieks, Krievins: "Diagnostic Value of Circulating CXC Chemokines in Non-small Cell Lung Cancer." dans: Anticancer research, Vol. 35, Issue 12, pp. 6979-83, (2016) (PubMed).

Hüfner, Koudouovoh-Tripp, Kandler, Hochstrasser, Malik, Giesinger, Semenitz, Humpel, Sperner-Unterweger: "Differential changes in platelet reactivity induced by acute physical compared to persistent mental stress." dans: Physiology & behavior, Vol. 151, pp. 284-91, (2015) (PubMed).

Leszczak, Popat: "Improved in vitro blood compatibility of polycaprolactone nanowire surfaces." dans: ACS applied materials & interfaces, Vol. 6, Issue 18, pp. 15913-24, (2014) (PubMed).

Popescu, Codrici, Albulescu, Mihai, Enciu, Albulescu, Tanase: "Potential serum biomarkers for glioblastoma diagnostic assessed by proteomic approaches." dans: Proteome science, Vol. 12, Issue 1, pp. 47, (2014) (PubMed).

Ollivier, Syvannarath, Gros, Butt, Loyau, Jandrot-Perrus, Ho-Tin-Noé: "Collagen can selectively trigger a platelet secretory phenotype via glycoprotein VI." dans: PLoS ONE, Vol. 9, Issue 8, pp. e104712, (2014) (PubMed).

Thasneem, Rekha, Sajeesh, Sharma: "Biomimetic mucin modified PLGA nanoparticles for enhanced blood compatibility." dans: Journal of colloid and interface science, Vol. 409, pp. 237-44, (2014) (PubMed).

Thasneem, Sajeesh, Sharma: "Glucosylated polymeric nanoparticles: a sweetened approach against blood compatibility paradox." dans: Colloids and surfaces. B, Biointerfaces, Vol. 108, pp. 337-44, (2013) (PubMed).

Tillack, Naegele, Haueis, Schippling, Wandinger, Martin, Sospedra: "Gender differences in circulating levels of neutrophil extracellular traps in serum of multiple sclerosis patients." dans: Journal of neuroimmunology, Vol. 261, Issue 1-2, pp. 108-19, (2013) (PubMed).

Zabini, Nagaraj, Stacher, Lang, Nierlich, Klepetko, Heinemann, Olschewski, Bálint, Olschewski: "Angiostatic factors in the pulmonary endarterectomy material from chronic thromboembolic pulmonary hypertension patients cause endothelial dysfunction." dans: PLoS ONE, Vol. 7, Issue 8, pp. e43793, (2012) (PubMed).

Aneja, Jalagadugula, Mao, Singh, Rao: "Mechanism of platelet factor 4 (PF4) deficiency with RUNX1 haplodeficiency: RUNX1 is a transcriptional regulator of PF4." dans: Journal of thrombosis and haemostasis : JTH, Vol. 9, Issue 2, pp. 383-91, (2011) (PubMed).

Détail du antigène

Antigène: PF4 (Platelet Factor 4 (PF4))

Autre désignation: PF4 (PF4 Produits)

Synonymes: CXCL4 Kit ELISA, PF-4 Kit ELISA, SCYB4 Kit ELISA, Cxcl4 Kit ELISA, Scyb4 Kit ELISA, Pf4a Kit ELISA, RATPF4A Kit ELISA, PF4A Kit ELISA, CXCL4L1 Kit ELISA, CXCL4V1 Kit ELISA, PF4-ALT Kit ELISA, SCYB4V1 Kit ELISA, PF4 Kit ELISA, platelet factor 4 Kit ELISA, platelet factor 4 variant 1 Kit ELISA, PF4 Kit ELISA, Pf4 Kit ELISA, LOC703451 Kit ELISA, PF4V1 Kit ELISA, LOC100524373 Kit ELISA, LOC102181854 Kit ELISA, LOC100630489 Kit ELISA

Sujet: The Human PF-4 (Platelet Factor-4) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human PF-4 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human PF-4 coated on a 96-well plate. Standards and samples are pipetted into the wells and PF-4 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human PF-4 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of PF-4 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.

ID gène: 9547

UniProt: O95715