G-CSF IQ-ELISA Kit

Catalog No. ABIN6244356

This qPCR ELISA kit is designed for the quantitative measurement of Human G-CSF.

Quick Overview for G-CSF IQ-ELISA Kit (ABIN6244356)

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

Reactivity: Human

Detection Method: qPCR

Method Type: Sandwich ELISA

Detection Range: 0.0512 pg/mL - 800 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma, Serum

Product Details G-CSF IQ-ELISA Kit

Minimum Detection Limit: 0.0512 pg/mL

Purpose: Human Immunoquantitative (PCR-Based) GCSF ELISA Kit for cell culture supernatants, plasma, and serum samples.

Analytical Method: Semi-Quantitative

Characteristics: IQELISAs employ specific capture antibodies coated on a 96-well PCR plate. Standards and samples are pipetted into the wells; the target protein in the standards and samples binds to the immobilized antibody. The wells are washed and the detection affinity reagent is added to the wells where it binds to any captured antigen. The wells are washed, and primers and PCR master mix are added to each well. The plate is placed into a real time PCR instrument for cycling and measurement of DNA amplification. The cycle number where amplification is detected is proportional to the amount of affinity detection reagent that bound to captured antigen in each well.

Components:

• GCSF Microplate (Item A): 96 well PCR plate coated with anti-Human GCSF
• Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution
• Standards (Item C): 2 vials of recombinant Human GCSF
• Assay Diluent A (Item D): 30 ml diluent buffer, 0.09% sodium azide as preservative. For Standard/Sample (serum/plasma) diluent
• Assay Diluent B (Item E): 15 ml of 5x concentrated buffer. For Standard/Sample (cell culture medium/urine) diluent
• Detection Affinity Reagent for GCSF (Item F): 2 vials of a 4x concentrated solution of anti-Human GCSF affinity reagent
• IQELISA Detection Reagent (Item G): 1.4ml of a 10x concentrated stock
• Primer Solution (Item I): 1.7 ml vial
• PCR Master Mix (Item J): 1.2 ml vial
• PCR Preparation buffer (Item K): 1ml vial of 10x concentrated buffer
• Final Wash Buffer (Item L): 10ml vial of 10x concentrated buffer

Material not included:

• Real-time PCR instrument, Bio-Rad recommended
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Absorbent paper
• Distilled or deionized water
• Log-log graph paper or computer and software for data analysis
• Tubes to prepare standard or sample dilutions
• Heating block or water bath capable of 80^°C

Application Details

Application Notes: The Immuno-Quantitative ELISA (IQELISA) kits are an innovative assay platform that combines the specificity and ease of use of an ELISA with the sensitivity of real-time PCR. This results in an assay that is simultaneously familiar and cutting edge and enables the use of only 1/10th the sample volume while also providing 10x more sensitivity than a traditional ELISA.

Comment:

Features:
- Excellent for follow up/confirmation of antibody array data
- Pre-coated 96 well plate
- Strip plate format minimizes waste
- Complete kit includes all necessary reagents
- 10x more sensitive than ELISA
- Reproducible and reliable
- Optimized for serum, plasma, urine and cell culture medium
- 1/10th the sample volume of an ELISA

Research Applications:
- Detection of quantitative protein levels in biological fluids
- Validation of antibody array results
- Validation of biomarker discovery studies

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards as instructed
2. Add 25 μL standard or sample to each well. Incubate for 2.5 hours at room temperature or overnight at 4^°C
3. Add 25 μL detection affinity reagent to each well. Incubate 1 hour at room temperature
4. Add 25μL of IQELISA Detection Reagent to each well. Incubate 1 hour
5. Add 15μL Primer solution 10μL of PCR master mix to each well
6. Run real-time PCR

Reagent Preparation:

Recommended Dilution for serum and plasma samples2-10 fold

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: May be stored for up to 6 months at 2° to 8°C from the date of shipment. Standard (recombinant protein) should be stored at -20°C or -80°C (recommended at -80°C) after reconstitution. Opened PCR plate or reagents may be stored for up to 1 month at 2° to 8°C. Note: the kit can be used within one year if the whole kit is stored at -20°C. Avoid repeated freeze-thaw cycles.

Expiry Date: 6 months

Target Details

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

Alternative Name: GCSF

Gene ID: 1440

UniProt: P09919

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