Lipoxin B4 ELISA Kit

Catalog No. ABIN5519077

Product Details

Target: Lipoxin B4

Reactivity: Various Species

Detection Method: Colorimetric

Method Type: Competition ELISA

Detection Range: 0.313-20 ng/mL

Minimum Detection Limit: 0.313 ng/mL

Application: ELISA

Purpose: The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.

Sample Type: Plasma, Serum

Analytical Method: Quantitative

Sensitivity: 0.188 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Reference Standard & Sample Diluent
• Biotinylated Detection Antibody (100 x concentrate)
• HRP Conjugate (100 x concentrate)
• Biotinylated Detection Antibody Diluent
• HRP Conjugate Diluent
• Substrate Reagent
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Application Details

Sample Volume: 50µL

Plate: Pre-coated

Protocol:

1. Add 50 µL standard or sample to each well. Immediately add 50 µL Biotinylated Detection Antibody to each well. Incubate for 45 min at 37 °C.
2. Aspirate and wash 3 times.
3. Add 100 µL HRP Conjugate to each well. Incubate for 30 min at 37 °C.
4. Aspirate and wash 5 times.
5. Add 90 µL Substrate Reagent. Incubate 15 min at 37 °C.
6. Add 50 µL Stop Solution. Read at 450 nm immediately.
7. Calculation of results.

Reagent Preparation:

1. Bring all reagents to room temperature (18~25 °C) before use. Follow the Microplate reader manual for set-up and preheat it for 15 min before OD measurement.
2. Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer. Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved.
3. Standard working solution: Centrifuge the standard at 10,000xg for 1 min. Add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 10 min and invert it gently several times. After it dissolves fully, mix it thoroughly with a pipette. This reconstitution produces a working solution of 20 ng/mL. Then make serial dilutions as needed. The recommended dilution gradient is as follows: 20, 10, 5, 2.5, 1.25, 0.63, 0.31, 0 ng/mL. Dilution method: Take 7 EP tubes, add 500 μLof Reference Standard & Sample Diluent to each tube. Pipette 500 μLof the 20 ng/mL stock solution to the first tube and mix up to produce a 10 ng/mL working solution. Pipette 500 μLof the solution from the former tube into the latter one according to these steps. The illustration below is for reference. Note: the last tube is regarded as a blank. Don't pipette solution into it from the former tube.
4. Biotinylated Detection Antibody working solution: Calculate the required amount before the experiment (50 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the stock tube before use, dilute the 100x Concentrated Biotinylated Detection Antibody to 1xworking solution with Biotinylated Detection Antibody Diluent.
5. Concentrated HRP Conjugate working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Dilute the 100x Concentrated HRP Conjugate to 1x working solution with Concentrated HRP Conjugate Diluent.

Restrictions: For Research Use only

Handling

Storage: 4 °C/-20 °C

Storage Comment: The unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the conditions since the kit is received.

Target Details

Target: Lipoxin B4

Alternative Name: LXB4(Lipoxin B4)