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ICAM1 Kit ELISA

ICAM1 Reactivité: Humain Colorimetric Sandwich ELISA 150 pg/mL-20 ng/mL Cell Culture Supernatant, Plasma, Serum
N° du produit ABIN625087
  • Antigène Voir toutes ICAM1 Kits ELISA
    ICAM1 (Intercellular Adhesion Molecule 1 (ICAM1))
    Reactivité
    • 9
    • 8
    • 7
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    150 pg/mL-20 ng/mL
    Seuil minimal de détection
    150 pg/mL
    Application
    ELISA
    Fonction
    Human ICAM-1 (CD54) 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 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-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, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC
    Sensibilité
    150 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
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  • Indications d'application
    Recommended Dilution for serum and plasma samples5 - 200 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 A (Item D) should be used for dilution of serum/plasma samples. Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 5-200 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 should be diluted 5-fold with deionized or distilled water.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 750 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 20 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Pipette 250 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the 20 ng/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 ng/mL). The 20 ng/mL standard in Assay Diluent B may be saturated, we recommend 10 ng/mL serves as starting point (the highest standard point) for Assay Diluent B. 250 µL Standard, Item C + 750 µL 250myl 250 µL 250 µL 250 µL 250 µL 20 10 5 2.5 1.25 0.625 0.313 0 ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL 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 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 340-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 25 µL of HRP-Streptavidin concentrate into a tube with 8.5 ml 1x Assay Diluent B to prepare a 340-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 A Human sICAM-1 concentration (ng/mL) O D =4 50 n m 0.01 0.1 1 10 0.1 1 10 100 Assay Diluent B Human sICAM-1 concentration (ng/mL) O D =4 50 n m 0.01 0.1 1 10 0.1 1 10 100
    Sensitivity: The minimum detectable dose of sICAM-1 is typically less than 150 pg/mL.
    Recovery: Recovery was determined by spiking various levels of human sICAM-1 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 97.48 88-106 Plasma 98.67 90-107 Cell culture media 97.37 89-107
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 98 97 96 Range ( %) 91-106 89-105 89-106 1:4 Average % of Expected 97 95 96 Range ( %) 89-106 88-105 90-107
    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
  • 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
  • Wu, Ding, Han, Arriens, Wei, Han, Pedroza, Jiang, Anolik, Petri, Sanz, Saxena, Mohan: "Antibody-Array-Based Proteomic Screening of Serum Markers in Systemic Lupus Erythematosus: A Discovery Study." dans: Journal of proteome research, Vol. 15, Issue 7, pp. 2102-14, (2016) (PubMed).

    Tawaramoto, Kaneto, Hashiramoto, Kawasaki, Tatsumi, Shimoda, Kamei, Matsuki, Mune, Kaku et al.: "Azelnidipine, but not amlodipine, reduces urinary albumin excretion and carotid atherosclerosis in subjects with type 2 diabetes: blood pressure control with olmesartan and azelnidipine in Type 2 ..." dans: Diabetology & metabolic syndrome, Vol. 7, pp. 80, (2015) (PubMed).

    Nirala, Gohil: "Effect of garlic component s-allyl cysteine sulfoxide on glycated human serum albumin induced activation of endothelial cells: an in vitro study." dans: European review for medical and pharmacological sciences, Vol. 19, Issue 11, pp. 2125-31, (2015) (PubMed).

    Zhang, Lin, Jiang, Xu, Luo, Mo, Li, Chen: "Extensive serum biomarker analysis in patients with ST segment elevation myocardial infarction (STEMI)." dans: Cytokine, Vol. 76, Issue 2, pp. 356-62, (2015) (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." dans: Journal of hypertension, Vol. 31, Issue 2, pp. 384-92, (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." dans: Cell adhesion & migration, Vol. 7, Issue 2, pp. 237-45, (2013) (PubMed).

    Benagiano, Munari, Ciervo, Amedei, Paccani, Mancini, Ferrari, Della Bella, Ulivi, DElios, Baldari, Prisco, de Bernard, DElios: "Chlamydophila pneumoniae phospholipase D (CpPLD) drives Th17 inflammation in human atherosclerosis." dans: Proceedings of the National Academy of Sciences of the United States of America, Vol. 109, Issue 4, pp. 1222-7, (2012) (PubMed).

    Bala, Gohil: "Interaction of glycated protein and DFO mimicked hypoxia in cellular responses of HUVECs." dans: Molecular bioSystems, Vol. 8, Issue 10, pp. 2657-63, (2012) (PubMed).

    Jang, Choi, Kim, Shin, Lee, Han: "Reactivation of transgene expression by alleviating CpG methylation of the Rous sarcoma virus promoter in transgenic quail cells." dans: Molecular biotechnology, Vol. 49, Issue 3, pp. 222-8, (2012) (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." dans: Molecular and cellular biochemistry, Vol. 359, Issue 1-2, pp. 73-81, (2011) (PubMed).

    Rudofsky, Roeder, Merle, Hildebrand, Nawroth, Wolfrum: "Weight loss improves endothelial function independently of ADMA reduction in severe obesity." dans: Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et métabolisme, Vol. 43, Issue 5, pp. 343-8, (2011) (PubMed).

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    Springer: "Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm." dans: Cell, Vol. 76, Issue 2, pp. 301-14, (1994) (PubMed).

  • Antigène Voir toutes ICAM1 Kits ELISA
    ICAM1 (Intercellular Adhesion Molecule 1 (ICAM1))
    Autre désignation
    ICAM-1 (ICAM1 Produits)
    Synonymes
    BB2 Kit ELISA, CD54 Kit ELISA, P3.58 Kit ELISA, Icam-1 Kit ELISA, Ly-47 Kit ELISA, MALA-2 Kit ELISA, ICAM Kit ELISA, ICAM-1 Kit ELISA, ICAM1 Kit ELISA, intercellular adhesion molecule 1 Kit ELISA, intercellular adhesion molecule-1 Kit ELISA, ICAM1 Kit ELISA, Icam1 Kit ELISA, ICAM-1 Kit ELISA
    Classe de substances
    Viral Protein
    Sujet
    SICAM-1 (soluble Intercellular Adhesion Molecular-1) has been reported in serum, cerebrospinal fluid and bronchoalveolar lavage. ICAM-1 expression is weak on leukocytes, epithelial and resting endothelial cells. ICAM-1 is a ligand for LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18). Its expression is up-regulated upon stimulation by inflammatory mediators such as cytokines and LPS. The Human sICAM-1 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human soluble ICAM-1 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human sICAM-1 coated on a 96-well plate. Standards and samples are pipetted into the wells and sICAM-1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human sICAM-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 sICAM-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%.
    ID gène
    3383
    UniProt
    P05362
    Pathways
    Cellular Response to Molecule of Bacterial Origin, Regulation of Actin Filament Polymerization, Carbohydrate Homeostasis, Regulation of Leukocyte Mediated Immunity, Thromboxane A2 Receptor Signaling
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