Sonic Hedgehog IQ-ELISA Kit

Produktnummer ABIN6730851

Human Sonic Hedgehog ELISA-Kit qPCR-Assay zur Quantifizierung von Human Sonic Hedgehog.

Kurzübersicht für Sonic Hedgehog IQ-ELISA Kit (ABIN6730851)

Target: Sonic Hedgehog (SHH)

Reaktivität: Human

Nachweismethode: qPCR

Methodentyp: DNA-Binding ELISA

Applikation: ELISA

Proben: Cell Culture Supernatant, Plasma, Serum

Produktdetails Sonic Hedgehog IQ-ELISA Kit

Verwendungszweck: Human Immunoquantitative (PCR-Based) Sonic Hedgehog N-Terminal ELISA Kit for cell culture supernatants, plasma, and serum samples.

Analytische Methode: Quantitative

Spezifität: 4.2 pg/ml in Assay Diluent C, and8 pg/ml in Assay Diluent B. (Anticipated minimum sensitivity. Assay may detect lower levels of antigen.)

Sensitivität: 4.2 pg/mL

Produktmerkmale: The Immuno Quantitative Enzyme Linked ImumunoSorbent Assay (IQELISA™) is an innovative new assay 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 lower sample volumes while also providing more sensitivity. The Human Sonic Hedgehog N-Terminal IQELISA™ Kit is a modified ELISA assay with high sensitivity qPCR readout for the quantitative measurement of Human Sonic Hedgehog N-Terminal in serum, plasma, and cell culture supernatants. This assay employs an antibody specific for Human Sonic Hedgehog N-Terminal coated on a 96-well PCR plate. Standards and samples are pipetted into the wells and Sonic Hedgehog N-Terminal present in a sample is bound to the wells by the immobilized antibody. The wells are washed and a detection affinity molecule is added to the plates. After washing away unbound detection affinity molecule, primers and a PCR master mix are added to the wells and data is collected using qPCR. Ct values obtained from the qPCR are then used to calculate the amount of antigen contained in each sample, where lower Ct values indicate a higher concentration of antigen.

Bestandteile:

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

Benötigtes Material:

• 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

Anwendungsinformationen

Applikationshinweise: 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.

Kommentare:

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

Plattentyp: Pre-coated

Protokoll:

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

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Informationen zur Lagerung: 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°C to 8°C.

Haltbarkeit: 6 months

Antigendetails

Target: Sonic Hedgehog (SHH)

Andere Bezeichnung: Sonic Hedgehog

Gen-ID: 6469

UniProt: Q15465

Pathways: Hedgehog Signalweg, Dopaminergic Neurogenesis, Regulation of Muscle Cell Differentiation, Tube Formation, Skeletal Muscle Fiber Development