GLI3 IQ-ELISA Kit

Catalog No. ABIN6385958

Product Details GLI3 IQ-ELISA Kit

Target: GLI3 (GLI Family Zinc Finger 3 (GLI3))

Reactivity: Human

Detection Method: qPCR

Method Type: Sandwich ELISA

Application: ELISA

Purpose: Human Immunoquantitative (PCR-Based) GLI-3 ELISA Kit for cell culture supernatants, plasma, and serum samples.

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Semi-Quantitative

Sensitivity: 4 pg/mL

Characteristics: IQ-ELISAs 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:

• GLI-3 Microplate (Item A): 96 well PCR plate coated with anti-Human GLI-3
• Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution
• Standards (Item C): 2 vials of recombinant Human GLI-3
• 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 GLI-3 (Item F): 2 vials of a 4x concentrated solution of anti-Human GLI-3 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:

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. Also called immuno-PCR, this detection platform results in an assay that is simultaneously familiar and cutting edge. Compared to traditional ELISA, IQELISA™ enables the use of only 1/10th the sample volume while also providing 10x more sensitivity.

Sample Volume: 25 µL

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

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C,-80 °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: GLI3 (GLI Family Zinc Finger 3 (GLI3))

Alternative Name: GLI-3 (GLI3 Products)

Synonyms: ACLS IQ-ELISA Kit, GCPS IQ-ELISA Kit, GLI3-190 IQ-ELISA Kit, GLI3FL IQ-ELISA Kit, PAP-A IQ-ELISA Kit, PAPA IQ-ELISA Kit, PAPA1 IQ-ELISA Kit, PAPB IQ-ELISA Kit, PHS IQ-ELISA Kit, PPDIV IQ-ELISA Kit, xgli3 IQ-ELISA Kit, AI854843 IQ-ELISA Kit, AU023367 IQ-ELISA Kit, Bph IQ-ELISA Kit, Pdn IQ-ELISA Kit, Xt IQ-ELISA Kit, add IQ-ELISA Kit, im:7142603 IQ-ELISA Kit, GLI3 IQ-ELISA Kit, GLI family zinc finger 3 IQ-ELISA Kit, GLI family zinc finger 3 L homeolog IQ-ELISA Kit, GLI-Kruppel family member GLI3 IQ-ELISA Kit, GLI3 IQ-ELISA Kit, Gli3 IQ-ELISA Kit, gli3.L IQ-ELISA Kit, gli3 IQ-ELISA Kit

Gene ID: 2737

UniProt: P10071

Pathways: Hedgehog Signaling