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Home / ELISA Kit / PharmaTest ELISA Kits

Filgrastim (GCSF) Pharmacokinetic ELISA | EL-1611-73196

SKU : BHE18300019

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Mfr.No.	EL-1611-73196
Detection Range	9.375ng/mL - 500 ng/mL
Reactivity	Human
Assay Duration	2.5 hours
Assay Type	Direct, Sandwich
Description	Granulocyte colony-stimulating factor (G-CSF or GCSF), also known as colony-stimulating factor 3 (CSF 3), is a glycoprotein that stimulates the bone marrow to produce granulocytes and stem cells and release them into the bloodstream Reagent preparation Prepare only the appropriate amount of required reagent on the day of use. Store all reagents as per instructions stated on the label. 1. Wash Buffer (1X) Preparation: Dilute wash buffer concentrate with ultra-pure water 1/20 before use (for example add 25mL concentrate to 475mL ultrapure water). Mix well. 2. Secondary Antibody (1X) Preparation: Dilute secondary antibody with assay buffer 1/1000 before use (for example add 12ul to 12mL of assay buffer). Mix well. 3. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 12ul concentrate to 12ml of assay buffer). Mix well. 4. Calibrator Preparation: Dilute the calibrator from 1mg/ml down to 5ug/ml by pipetting 5uL of calibrator stock into 995uL assay buffer. Label "Cal. Int." Mix well. Prepare calibrators with concentrations ranging from 500 ng/ml to 9.375 ng/ml. The following is an example calibrator curve. Results calculation 1. Construct a standard curve by plotting the absorbance obtained from each standard against concentration. Use a 4 or 5 parameter curve fit. 2. The concentration of the unknowns can be back calculated directly from this standard curve using the absorbance value for each sample. 3. Any sample diluted more or less than the standard series will need additional data correction. For example, if the sample is diluted 1/50, then the concentration will be calculated by dividing by 2 due to the calibrators being 2 times more diluted. Similarly, if the sample is diluted 1/500, then the concentration will be calculated by multiplying by a correction factor of 5 due to the calibrators being 5 times more concentrated. Sample preparation Dilute calibrators and test samples 1/50 with assay buffer (for example add 5uL of prepared calibrator or sample to 245uL of assay buffer). Mix well. Do not store diluted samples.
Detection Method	Peroxidase / OD450
Kit Components	Coated microtiter plate, 96 wells (1x8 strips) 1 Calibrator diluent 1.8ml Calibrator (1mg/ml) 12?l 20X wash buffer 25ml Assay buffer 50ml 1000X secondary antibody 1000X detection reagent 17?l TMB 12ml TMB stop solution 12ml
Limit Of Detection	9.375 ng/mL
Precision	< 25% for ULOQ and LLOQ and <20% for the remaining concentrations
Notes	This kit is compatible with EDTA-plasma, heparinplasma and serum samples. Samples can be stored at or below -20°C for up to 1 year.
Usage Notes	granulocyte colony stimulating factor in biological matrices
Protocol	This assay employs the indirect sandwich enzyme immunoassay technique. Anti-GCSF is coated onto a 96 well microplate. Calibrator and test samples prepared by dilution into assay buffer and are pipetted into the appropriate wells. GCSF present in biological matrices is bound by the immobilized anti- GCSF antibody. After washing away any unbound substances, tagged anti-GCSF secondary antibody is added to the wells. After washing, diluted detection reagent is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of GCSF present in test samples. The color development is stopped and the intensity of the color is measured.

Gene ID	1440

Applications	ELISA

Storage	-20C, 1 year

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SKU	Supplier No.	Size	Your price	Quantity
BHE18300019	EL-1611-73196	1X96 wells	$809.99		Add to Cart

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