Human HLA-DMA ELISA Kit, Lot 21AO-1214 [Cancer Immune Checkpoint Assay Kit]

The kit is designed for in vitro quantitative measurement of Human HLA-DMA in Plasma, Serum, Tissue Homogenate.

This immunoassay kit allows for the in vitro quantitative determination of human HLA-DMA,HLA-DMAconcentrations in serum, plasma, tissue homogenates, cell culture supernates, and other biological fluids.

ELISA

The kit should not be used beyond the expiration date on the kit label. Do not mix or substitute reagents with those from other lots or sources. If samples generate values higher than the highest standard, further dilute the samples with the Sample Diluent and repeat the assay. Any variation in standard diluent, operator, pipetting technique, washing technique,incubation time or temperature, and kit age can cause variation in binding.

HLA-DMA

Human

Colorimetric

Competition ELISA

Quantitative

Plasma, Serum, Tissue Homogenate

This assay recognizes recombinant and natural Human HLA-DMA. No significant cross-reactivity or interference was observed.

Limited by current skills and knowledge, it is impossible for us to complete the cross- reactivity detection between human HLA-DMA and all the analogues, therefore, cross reaction may still exist.

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.