Human CD117 molecule (CD117) ELISA kit instruction manual - Database & Sql Blog Articles

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Human CD117 molecule (CD117) ELISA kit

User's Guide

Human CD117 molecule (CD117) ELISA kit

Kit Name: Human CD117 molecule (CD117) ELISA kit

Kit Use: Quantitative detection of CD117 molecule (CD117) content in serum, plasma, and related fluid samples.

Detection Principle: The kit uses a two-antibody sandwich ELISA method. After adding standard and sample to the pre-coated enzyme-labeled plate, unbound components are washed away. Then, substrates A and B are added sequentially. TMB is converted into a blue product under horseradish peroxidase (HRP), which turns yellow after acid treatment. The color intensity is directly proportional to the concentration of CD117 molecule (CD117) in the sample. The OD value is measured at 450 nm, and the sample concentration is calculated based on the standard curve.

Kit Composition:

1 Enzyme-labeled coated plate 12 wells × 8 pieces
2 Standard: 200 ng/ml 0.6 mL
3 Substrate Solution A 6 mL
4 20× concentrated washing solution 20 mL
5 Sample diluent 6 mL
6 Enzyme standard reagent 6 mL
7 Substrate Solution B 6 mL
8 Stop solution 6 mL
9 Instruction manual 1 copy
10 Sealing film 1 sheet
11 Sealed bag 1 piece

Notes: Dilute the standard with standard dilution to concentrations of 200, 100, 50, 25, 12.5, and 6.25 ng/mL.

Reagents and Equipment Needed but Not Provided:

  1. 37°C incubator or water bath
  2. Microplate reader
  3. Pipettes and disposable tips
  4. Distilled water
  5. Disposable test tubes
  6. Absorbent paper

Procedure:

  1. Preparation: Remove the kit from the refrigerator and let it reach room temperature for 30 minutes.
  2. Dilute the 20× washing solution with distilled water to the working concentration.
  3. Add standards and samples to the enzyme-labeled plate. Add 50 μL of standard to each standard well, 10 μL of sample and 40 μL of diluent to each sample well (diluted 5 times), and leave blank control wells empty.
  4. Incubate at 37°C for 30 minutes.

Sample Requirements:

  1. Do not contain sodium azide (NaN3), as it inhibits HRP activity.
  2. Extract samples immediately after collection and store at -20°C if not tested right away. Avoid repeated freezing and thawing.
  3. Ensure the sample is fully centrifuged and free of hemolysis and particles.

Precautions:

  1. Follow the manual strictly. Results must be based on microplate reader readings.
  2. The quantitative range is 0.1–200 ng/mL. If outside this range, use a special dilution solution and recalculate.
  3. If the color is too light, extend the substrate incubation time slightly.
  4. Substrate B is light-sensitive; keep it away from light.

Summary of Operating Procedures:

  1. Prepare reagents, samples, and standards.
  2. Add samples and standards, incubate at 37°C for 30 minutes.

Range of Detection: 0.1–200 ng/mL

Specification: 96T/box, 48T/box

Storage: 2–8°C, protected from light and moisture.

Validity Period: 6 months

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