CD74 ELISA Kit

Catalog No. ABIN5519135

The Human CD74 ELISA Kit (ABIN5519135) is a Colorimetric ELISA Kit designed to quantify Human CD74.

Quick Overview for CD74 ELISA Kit (ABIN5519135)

Target: CD74 (HLA-DR-gamma (CD74))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.63 ng/mL - 40 ng/mL

Application: ELISA

Sample Type: Plasma, Serum

Product Details CD74 ELISA Kit

Minimum Detection Limit: 0.63 ng/mL

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

Analytical Method: Quantitative

Sensitivity: 0.38 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: 100 µL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

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

Reagent Preparation:

1. Equilibrate all reagents to room temperature (18-25 °C) before use. If the kit will not be used up in one assay, please only take out the necessary strips and reagents for present experiment, and store the remaining strips and reagents at required condition.
2. Standard Working Solution: Centrifuge the standard at 10,000x g for 1 min. Reconstitute with 1 mL of the Standard & Sample Diluent, allow to stand for 10 minutes, and gently invert several times until completely dissolved. Alternatively, vortex briefly at low speed, remove bubbles by low-speed centrifugation if necessary.This produces a 40 ng/mL stock solution. Prepare serial dilutions according to experimental requirements. Recommended dilution gradient: 40, 20, 10, 5, 2.5, 1.25, 0.63, 0 ng/mL Serial Dilution Procedure: Dispense 500 μL of Standard & Sample Diluent into 7 EP tubes, respectively. Transfer 500 μL of the stock solution into the first tube. Sequentially transfer 500 μL from the preceding tube into the next tube for subsequent dilutions. Designate the last tube as the blank, do not transfer any solution into it. Aliquot the stock and store at -20 ° C. Use within two weeks and avoid repeated freeze-thaw cycles. Serially diluted standards should be prepared immediately, prior to use.
3. Biotinylated Detection Ab working solution: Calculate the required volume (100 μL/well) plus a slight excess. Centrifuge the Concentrated Biotinylated Detection Antibody (100x) at 800 x g for 1 minute. Dilute to 1x with Biotinylated Detection Ab Diluent at a ratio of 1:99. Prepare the working solution immediately, prior to use.
4. HRP Conjugate Working Solution: Calculate the required volume (100 μL/well) plus a slight excess. Centrifuge the Concentrated HRP Conjugate (100x) at 800 x g for 1 minute. Centrifuge the Concentrated HRP Conjugate (100x) at 800 x g for 1 minute. Prepare the working solution immediately, prior to use.
5. Wash Buffer: Prepare the working Wash Buffer by diluting 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water yielding 750 mL. 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. For same day use only.

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 for CD74

Target: CD74 (HLA-DR-gamma (CD74))

Alternative Name: CD74(HLA class II histocompatibility antigen gamma chain)

UniProt: P04233

Pathways: Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Negative Regulation of intrinsic apoptotic Signaling, Cancer Immune Checkpoints