Human HLA-DMA ELISA Kit, Lot 21AO-1214 [Cancer Immune Checkpoint Assay Kit] (CAT#: IOK-05-P1125)

Assay plate
Standard
HRP-avidin (100 x concentrate)
Biotin-antibody (100 x concentrate)
Sample Diluent
HRP-avidin Diluent
Biotin-antibody Diluent
Wash Buffer (25 x concentrate)
TMB Substrate
Stop Solution
Adhesive Strip

Microplate reader.
Pipettes and pipette tips.
EP tube Deionized or distilled water.

100 μL

3 - 5 h

Pre-coated

1.Bring all reagents to room temperature (18-25 °C) before use.
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.
3.Standard working solution: 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 200pg/mL.
4.Biotinylated Detection Ab working solution: Centrifuge the Concentrated Biotinylated Detection Ab at 800xg for 1 min, then dilute the 100x Concentrated Biotinylated Detection Ab to 1x working solution with Biotinylated Detection Ab Diluent.
5.HRP Conjugate working solution: Calculate the required amount before the experiment.

1.Add 100µL of standard or sample per well. Cover with the adhesive strip provided. Incubate for 2 hours at 37°C.
2.Remove the liquid of each well, don't wash.
3.Add 100µL of Biotin-antibody (1×) to each well. Cover with a new adhesive strip. Incubate for 1 hour at 37°C.
4.Aspirate each well and wash, repeating the process two times for a total of three washes.After the last wash, remove any remaining wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
5.Add 100µL of HRP-avidin (1×) to each well. Cover the microtiter plate with a new adhesive strip. Incubate for 1 hour at 37°C.
6.Repeat the aspiration/wash process for five times as in step 6.
7.Add 90µL of TMB Substrate to each well. Incubate for 20 minutes at 37°C. Protect from light.
8.Add 50µL of Stop Solution to each well, gently tap the plate to ensure thorough mixing.
9.Determine the optical density of each well within 5 minutes using a microplate reader set to 450nm.

Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit.

The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

4 °C

The Assay Plate, Standard, Detection Reagent A and Detection Reagent B should be stored at -20°C upon being received. After receiving the kit , Substrate should be always stored at 4°C.

6 months

The Assay Plate, Standard, Detection Reagent A and Detection Reagent B should be stored at -20°C upon being received. After receiving the kit , Substrate should be always stored at 4°C.

For Research Use only

Major Histocompatibility Complex, Class II, DM alpha

RT1.Ma; RT1.DMa; HLA-DMA; H-2Ma; H2-Ma; D6S222E; DMA; HLADM; RING6; RT1 class II; locus DMa; SLA-DM alpha chain; major histocompatibility complex; class II; DM alpha; histocompatibility 2; class II; locus DMa; RT1-DMa; SLA-DMA; DLA-DMA; H2-DMa; HLA-DMA

Synonyms: HLA-DMA, MHC class II antigen DMA,Really interesting new gene 6 protein

P28067

Cancer Immune Checkpoints, Human Leukocyte Antigen (HLA) in Adaptive Immune Response

The microtiter plate provided in this kit has been pre-coated with an antibody specific to HLA-DMA, During the reaction, HLA-DMA in the sample or standard competes with a fixed amount of biotin-labeled HLA-DMA for sites on a pre-coated Monoclonal antibody specific to HLA-DMA. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.