G-CSF ELISA Kit

Catalog No. ABIN8118914

Human G-CSF ELISA Kit, Colorimetric assay for quantification of Human G-CSF.

Quick Overview for G-CSF ELISA Kit (ABIN8118914)

Target: G-CSF (CSF3) (Colony Stimulating Factor 3 (Granulocyte) (CSF3))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 9.375 ng/mL - 500 ng/mL

Application: ELISA

Sample Type: Plasma, Serum

Product Details G-CSF ELISA Kit

Minimum Detection Limit: 9.375 ng/mL

Purpose: granulocyte colony stimulating factor in biological matrices

Analytical Method: Quantitative

Specificity: GCSF

Components:

• Coated microtiter plate, 96 wells
• (1x8 strips) 1 Calibrator diluent 1.8ml
• Calibrator (1mg/ml) 12μl 20X wash buffer 25ml
• Assay buffer 50ml
• 1000X secondary antibody 1000X detection reagent 17μl TMB 12ml
• TMB stop solution 12ml

Material not included:

• Precision pipettes calibrated to deliver 5-1000μL
• Multi-channel pipette calibrated to deliver 50-200μL
• Plate shaker
• Disposable tips
• Vortex-Mixe
• Distilled or de-ionized water
• Microplate reader capable of reading 450nm with background subtraction

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 15 μL

Assay Time: 2.5 h

Plate: Pre-coated

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.

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 25 mL concentrate to 475 mL ultrapure water). Mix well. 2. Secondary Antibody (1X) Preparation: Dilute secondary antibody with assay buffer 1/1000 before use (for example add 12 μL to 12 mL of assay buffer). Mix well. 3. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 12 μL concentrate to 12 mL of assay buffer). Mix well. 4. Calibrator Preparation: Dilute the calibrator from 1 mg/mL down to 5 μg/mL by pipetting 5μL of calibrator stock into 995μL 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.

Sample Collection: 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.

Sample Preparation:

Dilute calibrators and test samples 1/50 with assay buffer (for example add 5μL of prepared calibrator or sample to 245μL of assay buffer). Mix well. Do not store diluted samples.

Calculation of Results:

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.

Assay Precision: < 25% for ULOQ and LLOQ and <20% for the remaining concentrations

Restrictions: For Research Use only

Handling

Preservative: Without preservative

Storage: -20 °C

Storage Comment: -20C, 1 year

Expiry Date: 12 months

Target Details for G-CSF

Target: G-CSF (CSF3) (Colony Stimulating Factor 3 (Granulocyte) (CSF3))

Alternative Name: granulocyte colony stimulating factor

Background: 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

Gene ID: 1440

Pathways: Cellular Response to Molecule of Bacterial Origin, Regulation of Actin Filament Polymerization