Porcine Apoptosis Inducing Factor (AIF) ELISA Kit Instructions - Database & Sql Blog Articles

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Kaixin micro test

Test probe P100-M3

Brand AVX TPSE226M035R0125 Low impedance tantalum capacitor AVX 22

Porcine Apoptosis-Inducing Factor (AIF) ELISA Kit

This kit is for research use only. It is not intended for diagnostic or therapeutic purposes.

Experimental Principle:

The level of porcine apoptosis-inducing factor (AIF) in the sample is determined using a double-antibody sandwich ELISA method. The microwell plate is pre-coated with a specific monoclonal antibody against AIF. After incubation with the sample, the AIF binds to the immobilized antibody. Then, an HRP-labeled secondary antibody is added, forming a complex of antibody-AIF-enzyme-labeled antibody. Following wash steps, TMB substrate is added, and the reaction is stopped with a stop solution. The color change is proportional to the concentration of AIF in the sample. The absorbance is measured at 450 nm, and the concentration is calculated from a standard curve.

Kit Composition:

• 1. 30× Washing Solution – 20ml × 1 bottle
• 2. Enzyme Standard Reagent – 6ml × 1 bottle
• 3. Enzyme-Labeled Coating Plate – 12 wells × 8 strips
• 4. Sample Diluent – 6ml × 1 bottle
• 5. Reagent A – 6ml × 1 bottle
• 6. TMB Substrate B – 6ml × 1 bottle
• 7. Stop Solution – 6ml × 1 bottle
• 8. Standard (800pg/ml) – 0.5ml × 1 bottle
• 9. Standard Dilutions – 1.5ml × 1 bottle
• 10. Instruction Manual – 1 copy
• 11. Sealing Film – 2 sheets
• 12. Sealed Bag – 1

Sample Requirements:

• 1. Samples should be processed as soon as possible after collection. If not tested immediately, store at -20°C and avoid repeated freeze-thaw cycles.
• 2. Avoid using samples containing NaN3, as it may inhibit HRP activity.

Procedure:

1. Dilute the standard according to the provided chart to create a standard curve.
2. Add 50μl of standard and 50μl of sample diluent to each well, followed by 10μl of the sample (final dilution: 5×).

Calculation:

Plot the standard curve using OD values vs. concentrations. Use the regression equation to calculate the unknown sample concentration and multiply by the dilution factor.

Precautions:

• Allow the kit to reach room temperature before use. Store unopened enzyme reagents in a sealed bag.
• Concentrated washing solution may crystallize; warm it gently if needed.
• Use a pipette for accuracy. Limit loading time to 5 minutes per batch.
• Always run a standard curve and duplicate wells. If the OD value exceeds the highest standard, dilute the sample and adjust calculations accordingly.
• Do not reuse sealing films to prevent cross-contamination.
• Keep TMB away from light.
• Follow the manual strictly and rely on microplate reader results.
• Treat all waste as biohazardous material.
• Do not mix components from different batches.
• In case of conflict between Chinese and English manuals, the English version takes precedence.

Storage: 2–8°C

Shelf Life: 6 months