PYCARD ELISA Kit

Catalog No. ABIN7672140

This Colorimetric ELISA kit is designed for the quantitative measurement of Human PYCARD.

Quick Overview for PYCARD ELISA Kit (ABIN7672140)

Target: PYCARD (PYD and CARD Domain Containing (PYCARD))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 78.125 pg/mL - 5000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Lysate

Grade: QuickTest

Product Details PYCARD ELISA Kit

Minimum Detection Limit: 78.125 pg/mL

Purpose: Human PYCARD (Apoptosis-associated speck-like protein containing a CARD) QuickTest ELISA Kit

Brand: QuickTest

Analytical Method: Quantitative

Sensitivity: 46.875 pg/mL

Components:

• ELISA Microplate (Dismountable)
• Lyophilized Standard
• Capture/Detection Antibody
• HRP-Streptavidin (orange)
• TMB Substrate
• Sample Dilution Buffer (blue)
• Stop Solution
• Wash Buffer (25x)
• Plate Sealer
• Product Description

Material not included:

• Microplate reader (wavelength: 450nm)
• 37 °C incubator (CO2 incubator for cell culture is not recommenced.)
• Automated plate washer or multi-channel pipette/5 mL pipettor (for manual washing purpose)
• Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips (Calibration is required before use.)
• Sterile tubes and sample tubes with disposable tips
• Absorbent paper and loading slot
• Deionized or distilled water

Application Details

Comment:

Traditional sandwich ELISA assay usually takes 3-4 hours. Most detection reagents in the market are concentrated and can't be distinguished from the colour. Dilution is required before use. We developed QuickTest ELISA kits to decrease customers’ operational errors, improve user's experience and get accurate assay result more efficiently. Compared with regular ELISA kits, QuickTest ELISA kits have more advantages in the same linearity, stability, recovery and storage conditions.

Sample Volume: 50 µL

Assay Time: 2 h

Plate: Pre-coated

Protocol: Capture antibody was conjugated to an affinity tag that was recognized by a specific antibody pre-coated on the QuickTest plate. Add the Cap/Det Ab working solution into each well, then add the standards and pilot samples into relevant wells. Mix thoroughly and then incubate. If the sample contains detection target, a capture antibody-detection target-biotin-detection antibody complex was formed. After washing off unbound conjugates, HRP-Streptavidin was added to incubate. Repeat the washing process and then TMB substrates were added to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm with a microplate reader. The concentration of the detection target in the sample was calculated by plotting a standard curve. The concentration of the target substance is proportional to the OD450 value.

Assay Procedure:

1. Take out the required strips, add 50 µL Cap/Det Ab into each well, then add 50 µL Standard or Sample into relevant well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure complete mixing then statically incubate for 60 minutes at 37 °C.
2. Wash the plate twice without immersion.
3. Add 100 µL HRP-Streptavidin (orange) into each well, seal the plate and statically incubate for 30 minutes at 37 °C.
4. Wash the plate five times without immersion.
5. Add 90 µL TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37 °C (Accurate TMB visualization control is required.).
6. Add 50 µL stop solution. Read at 450nm immediately and calculate.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: 2-8 °C for 12 months

Expiry Date: 12 months

Target Details for PYCARD

Target: PYCARD (PYD and CARD Domain Containing (PYCARD))

Alternative Name: PYCARD

Background: Apoptosis-associated speck-like protein containing a CARD|hASC|Caspase recruitment domain-containing protein 5|PYD and CARD domain-containing protein|Target of methylation-induced silencing 1|PYCARD|ASC|CARD5|TMS1

UniProt: Q9ULZ3

Pathways: Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Regulation of Actin Filament Polymerization, Positive Regulation of Endopeptidase Activity, Activated T Cell Proliferation, Inflammasome