Dihydrofolate Reductase IQ-ELISA Kit

Catalog No. ABIN6385265

Product Details Dihydrofolate Reductase IQ-ELISA Kit

Target: Dihydrofolate Reductase (DHFR)

Reactivity: Human

Detection Method: qPCR

Method Type: Sandwich ELISA

Application: ELISA

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

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Semi-Quantitative

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:

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

Alternative Name: DHFR (DHFR Products)

Synonyms: DHFR IQ-ELISA Kit, F10_orf160 IQ-ELISA Kit, ECK0049 IQ-ELISA Kit, JW0047 IQ-ELISA Kit, tmrA IQ-ELISA Kit, cb595 IQ-ELISA Kit, zgc:86637 IQ-ELISA Kit, DDBDRAFT_0187132 IQ-ELISA Kit, DDBDRAFT_0230113 IQ-ELISA Kit, DDB_0187132 IQ-ELISA Kit, DDB_0230113 IQ-ELISA Kit, CG14887 IQ-ELISA Kit, Dmel\\CG14887 IQ-ELISA Kit, dhfr IQ-ELISA Kit, DHFRP1 IQ-ELISA Kit, DYR IQ-ELISA Kit, 8430436I03Rik IQ-ELISA Kit, AA607882 IQ-ELISA Kit, AI662710 IQ-ELISA Kit, AW555094 IQ-ELISA Kit, Dhfr1 IQ-ELISA Kit, confers resistance to sulfonamides IQ-ELISA Kit, dihydrofolate reductase IQ-ELISA Kit, dihydrofolate reductase DhfR IQ-ELISA Kit, Dihydrofolate reductase IQ-ELISA Kit, dhfr IQ-ELISA Kit, DHFR IQ-ELISA Kit, dhfR IQ-ELISA Kit, folA IQ-ELISA Kit, dfrA IQ-ELISA Kit, LOC707382 IQ-ELISA Kit, CNJ01940 IQ-ELISA Kit, DDB_G0286755 IQ-ELISA Kit, Dhfr IQ-ELISA Kit, Mesil_0096 IQ-ELISA Kit, Arch_0408 IQ-ELISA Kit, Fbal_3367 IQ-ELISA Kit, Ilyop_0482 IQ-ELISA Kit, Lbys_2007 IQ-ELISA Kit, Riean_0733 IQ-ELISA Kit, Mrub_1855 IQ-ELISA Kit, Poras_0766 IQ-ELISA Kit, Halhy_3552 IQ-ELISA Kit, Mesop_3614 IQ-ELISA Kit, LOC609048 IQ-ELISA Kit

Gene ID: 1719

UniProt: P00374

Pathways: Mitotic G1-G1/S Phases