Transferrin Kit ELISA

N° du produit ABIN5564633

Détail du produit Transferrin Kit ELISA

Antigène: Transferrin (TF)

Reactivité: Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 0.625-20 ng/mL

Seuil minimal de détection: 0.625 ng/mL

Application: ELISA

Fonction: The AssayMax™ Mouse Transferrin ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for detection of mouse transferrin in plasma, serum, and cell culture samples. This assay employs a quantitative sandwich enzyme immunoassay technique that measures mouse transferrin in less than 4 hours. A polyclonal antibody specific for mouse transferrin has been pre-coated onto a 96-well microplate with removable strips. Transferrin in standards and samples is sandwiched by the immobilized antibody and biotinylated polyclonal antibody specific for mouse transferrin, which is recognized by a streptavidin-peroxidase conjugate. All unbound material is washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.

Marque: AssayMax™

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

Analytical Method: Quantitative

Ingrédients: Mouse Transferrin Microplate: A 96 well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against mouse transferrin. Sealing Tapes: Each kit contains 3 precut, pressure sensitive sealing tapes that can be cut to fit the format of the individual assay. Mouse Transferrin Standard: Mouse transferrin in a buffered protein base (40 ng, lyophilized). Biotinylated Mouse Transferrin Antibody (50x): A 50-fold concentrated biotinylated polyclonal antibody against mouse transferrin (140 l). MIX Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml). Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml, 2 bottles). Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (80 l). Chromogen Substrate: A ready-to-use stabilized peroxidase chromogen substrate tetramethylbenzidine (8 ml). Stop Solution: A 0.5 N hydrochloric acid to stop the chromogen substrate reaction (12 ml).

Matériel non inclus: Microplate reader capable of measuring absorbance at 405 nm. Pipettes (1-20 µL, 20-200 µL, and multiple channel). Deionized or distilled reagent grade water Incubator (37 °C)

Information d'application

Durée du test: 5 h

Plaque: Pre-coated

Protocole:

• Step 1. Add 50 μL of Standard or Sample per well. Incubate 2 hours.
• Step 2. Wash, then add 50 μL of Biotinylated Antibody per well. Incubate 1 hour.
• Step 3. Wash, then add 50 μL of SP Conjugate per well. Incubate 30 minutes.
• Step 4. Wash, then add 50 μL of Chromogen Substrate per well. Incubate 20 minutes.
• Step 5. Add 50 μL of Stop Solution per well. Read at 450 nm immediately.

Préparation des réactifs:

Freshly dilute all reagents and bring all reagents to room temperature before use. MIX Diluent Concentrate (10x): If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. 4 Dilute the MIX Diluent Concentrate 1:10 with reagent grade water. Store for up to 30 days at 2-8 °C.

Prélèvement de l'échantillon: Plasma: Collect plasma using one-tenth volume of 0.1 M sodium citrate as an anticoagulant. Centrifuge samples at 3000 x g for 10 minutes and use supernatants. Dilute samples 1:800000 with MIX Diluent or within the range of 1:160000 - 1:1600000, and assay. Depending on application needs, user should determine proper dilutions. The undiluted samples can be stored at -20 °C or below for up to 3 months. Avoid repeated freeze-thaw cycles (EDTA or Heparin can also be used as an anticoagulant). Serum: Samples should be collected into a serum separator tube. After clot formation, centrifuge samples at 3000 x g for 10 minutes, and remove serum. Dilute samples 1:800000 with MIX Diluent or within the range of 1:160000 - 1:1600000, and assay. Depending on application needs, user should determine proper dilutions. The undiluted samples can be stored at -20 °C or below for up to 3 months. Avoid repeated freeze-thaw cycles. Cell Culture Supernatants: Centrifuge cell culture media at 3000 x g for 10 minutes to remove debris. Collect supernatants and assay. Store samples at -20 °C or below. Avoid repeated freeze-thaw cycles. Refer to Sample Dilution Guidelines below for further instruction. Guidelines for Dilutions of 1:100 or Greater (for reference only, please follow the insert for specific dilution suggested) 1:100 1:10000 A) 4 μL sample: 396 μL buffer(100x) = 100 fold dilution Assuming the needed volume is less than or equal to 400 μL. A) 4 μL sample : 396 μL buffer (100x) B) 4 μL of A : 396 μL buffer (100x) = 10000 fold dilution Assuming the needed volume is less than or equal to 400 μL. 1:1000 1:100000 A) 4 μL sample : 396 μL buffer (100x) B) 24 μL of A : 216 μL buffer (10x) = 1000 fold dilution Assuming the needed volume is less than or equal to 240 μL. A) 4 μL sample : 396 μL buffer (100x) B) 4 μL of A : 396 μL buffer (100x) C) 24 μL of B : 216 μL buffer (10x) = 100000 fold dilution Assuming the needed volume is less than or equal to 240 μL.

Procédure de l'essai:

Prepare all reagents, standard solutions, and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature (20-25 °C). Remove excess microplate strips from the plate frame and return them immediately to the foil pouch with desiccants inside. Reseal the pouch securely to minimize exposure to water vapor and store in a vacuum desiccator. 5 Add 50 l of Mouse Transferrin Standard or sample per well. Cover wells with a sealing tape and incubate for 2 hours. Start the timer after the last addition. Wash five times with 200 l of Wash Buffer manually. Invert the plate each time and decant the contents, hit 4-5 times on absorbent material to completely remove the liquid. If using a machine, wash six times with 300 l of Wash Buffer and then invert the plate, decanting the contents, hit 4-5 times on absorbent material to completely remove the liquid. Add 50 l of Biotinylated Mouse Transferrin Antibody to each well and incubate for 1 hour. Wash the microplate as described above. Add 50 l of Streptavidin-Peroxidase Conjugate per well and incubate for 30 minutes. Turn on the microplate reader and set up the program in advance. Wash the microplate as described above. Add 50 l of Chromogen Substrate per well and incubate for 20 minutes or till the optimal blue color density develops. Gently tap plate to ensure thorough mixing and break the bubbles in the well with pipette tip. Add 50 l of Stop Solution to each well. The color will change from blue to yellow. Read the absorbance on a microplate reader at a wavelength of 450 nm immediately. If wavelength correction is available, subtract readings at 570 nm from those at 450 nm to correct optical imperfections. Otherwise, read the plate at 450 nm only. Please note that some unstable black particles may be generated at high concentration points after stopping the reaction for about 10 minutes, which will reduce the readings.

Calcul des résultats:

• Calculate the mean value of the duplicate or triplicate readings for each standard and sample.
• To generate a standard curve, plot the graph using the standard concentrations on the x-axis and the corresponding mean 450 nm absorbance (OD) on the y-axis. The best-fit line can be determined by regression analysis using log-log or four-parameter logistic curve-fit.
• Determine the unknown sample concentration from the standard curve and multiply the value by the dilution factor.

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: This product is for Research Use Only and is Not For Use In Diagnostic Procedures. Prepare all reagents (working diluent buffer, wash buffer, standard, biotinylated antibody, and SP conjugate) as instructed, prior to running the assay. Prepare all samples prior to running the assay. The dilution factors for the samples are suggested in this insert. However, the user should determine the optimal dilution factor. Spin down the SP conjugate vial and the biotinylated antibody vial before opening and using contents. The Stop Solution is an acidic solution. The kit should not be used beyond the expiration date. 2

Stock: 4 °C,-20 °C

Stockage commentaire: Upon arrival, immediately store components of the kit at recommended temperatures up to the expiration date. Store SP Conjugate and Biotinylated Antibody at -20°C. Store Microplate, Diluent Concentrate (10x), Wash Buffer, Stop Solution, and Chromogen Substrate at 2-8°C. Unused microplate wells may be returned to the foil pouch with the desiccant packs and resealed. May be stored for up to 30 days in a vacuum desiccator. Diluent (1x) may be stored for up to 30 days at 2-8°C. Store Standard at 2-8°C before reconstituting with Diluent and at -20°C after reconstituting with Diluent.

Détail du antigène

Antigène: Transferrin (TF)

Autre désignation: Transferrin (TF Produits)

Synonymes: ltf Kit ELISA, pro1557 Kit ELISA, pro2086 Kit ELISA, mgc107777 Kit ELISA, LOC692564 Kit ELISA, TF Kit ELISA, LOC100144362 Kit ELISA, tf Kit ELISA, LTF Kit ELISA, TFEW Kit ELISA, conalbumin Kit ELISA, tf-b Kit ELISA, MGC64306 Kit ELISA, PRO1557 Kit ELISA, PRO2086 Kit ELISA, TFQTL1 Kit ELISA, AI266983 Kit ELISA, Cd176 Kit ELISA, HP Kit ELISA, Tf Kit ELISA, Tfn Kit ELISA, hpx Kit ELISA, Trf Kit ELISA, cb285 Kit ELISA, gavi Kit ELISA, id:ibd3238 Kit ELISA, id:ibd3525 Kit ELISA, sb:cb285 Kit ELISA, wu:fb57g06 Kit ELISA, wu:fb62h02 Kit ELISA, wu:fb63h10 Kit ELISA, wu:fb64h10 Kit ELISA, zgc:112154 Kit ELISA, 143958_at Kit ELISA, CG6186 Kit ELISA, Dmel\\CG6186 Kit ELISA, TSF1 Kit ELISA, anon-EST:Posey265 Kit ELISA, tsf1 Kit ELISA, Pro-TRH Kit ELISA, IL-5 Kit ELISA, STF I Kit ELISA, TRF1 Kit ELISA, sTF1 Kit ELISA, sTf Kit ELISA, tf1 Kit ELISA, transferrin Kit ELISA, serotransferrin Kit ELISA, transferrin (ovotransferrin) Kit ELISA, transferrin L homeolog Kit ELISA, melanotransferrin Kit ELISA, transferrin-a Kit ELISA, Transferrin 1 Kit ELISA, thyrotropin releasing hormone Kit ELISA, interleukin 5 Kit ELISA, TF Kit ELISA, LOC477072 Kit ELISA, tf Kit ELISA, Tf Kit ELISA, LOC100144362 Kit ELISA, tf.L Kit ELISA, LOC5575625 Kit ELISA, Trf Kit ELISA, tfa Kit ELISA, Tsf1 Kit ELISA, TRH Kit ELISA, IL5 Kit ELISA, LOC101085148 Kit ELISA, LOC100726872 Kit ELISA, trf Kit ELISA

Sujet: Transferrin is a plasma protein that transports iron through the blood to the liver, spleen, and bone marrow (1).

ID gène: 22041

UniProt: Q921I1

Pathways: Transition Metal Ion Homeostasis