VCAM1 ELISA Kit

Produktnummer ABIN625106

Produktdetails VCAM1 ELISA Kit

Target: VCAM1 (Vascular Cell Adhesion Molecule 1 (VCAM1))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 0.3-60 ng/mL

Untere Nachweisgrenze: 0.3 ng/mL

Applikation: ELISA

Verwendungszweck: Human VCAM-1 (CD106) ELISA Kit for cell culture supernatants, plasma, and serum samples.

Proben: Plasma, Cell Culture Supernatant, Serum

Analytische Methode: Quantitative

Spezifität: This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiopoietin-1, Angiostatin , B7-1, BMP-7, CD14, CD30, CD40, CD40 Ligand, CTLA-4, CXCL16, Dkk-4, DR6, Endostatin, E-Selectin, Follistatin, HB-EGF, HVEM, ICAM-2, IGF-II, IL-10 Ra, IL-10 Rb, IL-18, IL-9 , IL-2 Ra, IL-2 Rb, IL-5 Ra, LAP, L-Selectin, MCSF R, MMP-1, 2, 3, 7, 8, 9, 10,12 and 13, PDGF-AB, SDF-1b, Tie-1, Tie- 2, TIMP-3.

Sensitivität: 300 pg/mL

Produktmerkmale:

• 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

Bestandteile:

• 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

Benötigtes Material:

• 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

Anwendungsinformationen

Applikationshinweise: Recommended Dilution for serum and plasma samples10 - 50 fold

Probenmenge: 100 μL

Plattentyp: Pre-coated

Protokoll:

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.

Aufbereitung der Reagenzien:

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 A (Item D) is used for dilution of serum/plasma samples, and Assay Diluent B (Item E) is used for dilution of culture supernatants and urine. 3. Assay Diluent B should be diluted 5-fold with deionized or distilled water. 4. Preparation of standard: Briefly spin the vial of Item C and then add 800 µl Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 100 ng/ml standard. Dissolve the powder thoroughly by a gentle mix. Add 300 µl VCAM-1 standard from the vial of Item C, into a tube with 200 µl Assay Diluent A or 1x Assay Diluent B to prepare a 60 ng/ml stock standard solution. Pipette 400 µl Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 ng/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 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 65-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) before use. HRP-Streptavidin concentrate should be diluted 16,000-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 2 µl of HRP-Streptavidin concentrate into a tube with 198.0 µl 1x Assay Diluent to prepare a 100-fold diluted HRP-Streptavidin solution (do not store the diluted solution for next day use). Mix through and then pipette 100 µl of prepared 100-fold diluted solution into a tube with 16 ml 1x Assay Diluent B to prepare a final 16,000 fold diluted HRP-Streptavidin solution.

Testdurchführung:

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 µl) 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 3. 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 3. 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.

Ergebnisberechnung:

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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: -20 °C

Informationen zur Lagerung: 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.

Haltbarkeit: 6 months

Referenzen für VCAM1 ELISA Kit (ABIN625106)

Fadel, Elshamaa, Nabhan, Essam, Kantoush, El Sonbaty, Raafat, Abd-El Haleem: "Soluble adhesion molecules as markers of native arteriovenous fistula thrombosis in children on uremia." in: Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis, Vol. 25, Issue 7, pp. 675-82, (2015) (PubMed).

Schlatzer, Sugalski, Chen, Barnholtz-Sloan, Davitkov, Hazlett, Funderburg, Rodriguez, Lederman, Sieg, Chance, Anthony: "Plasma proteome analysis reveals overlapping, yet distinct mechanisms of immune activation in chronic HCV and HIV infections." in: Journal of acquired immune deficiency syndromes (1999), Vol. 63, Issue 5, pp. 563-71, (2013) (PubMed).

Asato, Yoshida, Ogura, Nakama, Ishikawa, Nakao, Sassa, Enaida, Oshima, Ikeo, Gojobori, Kono, Ishibashi: "Comparison of gene expression profile of epiretinal membranes obtained from eyes with proliferative vitreoretinopathy to that of secondary epiretinal membranes." in: PLoS ONE, Vol. 8, Issue 1, pp. e54191, (2013) (PubMed).

Chao, Huang, Hsieh: "Inhibitive effect of purple sweet potato leaf extract and its components on cell adhesion and inflammatory response in human aortic endothelial cells." in: Cell adhesion & migration, Vol. 7, Issue 2, pp. 237-45, (2013) (PubMed).

Liu, Ho, Chen, Woo: "Effect of soy protein and isoflavones on blood pressure and endothelial cytokines: a 6-month randomized controlled trial among postmenopausal women." in: Journal of hypertension, Vol. 31, Issue 2, pp. 384-92, (2013) (PubMed).

Sadik, Mohamed, Ahmed: "The association of receptor of advanced glycated end products and inflammatory mediators contributes to endothelial dysfunction in a prospective study of acute kidney injury patients with sepsis." in: Molecular and cellular biochemistry, Vol. 359, Issue 1-2, pp. 73-81, (2011) (PubMed).

Tammali, Reddy, Srivastava, Ramana: "Inhibition of aldose reductase prevents angiogenesis in vitro and in vivo." in: Angiogenesis, Vol. 14, Issue 2, pp. 209-21, (2011) (PubMed).

Details zu VCAM1

Target: VCAM1 (Vascular Cell Adhesion Molecule 1 (VCAM1))

Andere Bezeichnung: VCAM-1 (VCAM1 Produkte)

Synonyme: CD106 ELISA Kit, INCAM-100 ELISA Kit, Vcam-1 ELISA Kit, VCAM1B ELISA Kit, fi76d06 ELISA Kit, wu:fi76d06 ELISA Kit, zgc:158875 ELISA Kit, VCAM1 ELISA Kit, vascular cell adhesion molecule 1 ELISA Kit, vascular cell adhesion molecule 1b ELISA Kit, VCAM1 ELISA Kit, Vcam1 ELISA Kit, vcam1b ELISA Kit, VCAM-1 ELISA Kit

Hintergrund: The Human VCAM-1 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human VCAM-1 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human VCAM-1 coated on a 96-well plate. Standards and samples are pipetted into the wells and VCAM-1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human VCAM-1 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 VCAM-1 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%.

Gen-ID: 7412

UniProt: P19320

Pathways: Carbohydrate Homeostasis