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Human 2,3-Disphosphoglycerate ELISA Kit

Catalogue No BSEK-163H
Species Human
Size 96 T | 48 T
Price POR
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SKU: 7881Hu Categories: ,
Human 2,3-Disphosphoglycerate ELISA Kit - Details
Sensitivity 0.1 nmol/ml
Range 0.625nmol/ml-40 nmol/ml
Sample Type use with serum, plasma, culture media, or any biological fluids
Storage 2-8℃
Detection Method Quantitative Detection

Human 2,3-Disphosphoglycerate ELISA Kit (Cat No: 7881Hu)

Sample preparation

  1. Serum preparation

After collection of the whole blood, allow the blood to clot by leaving it undisturbed at room temperature. This usually takes 10-20 minutes. Remove the clot by centrifuging at 2,000-3,000 rpm for 20 minutes. If precipitates appear during reservation, the sample should be centrifugated again.

  1. Plasma preparation

Collect the whole blood into tubes with anticoagulant (EDTA or citrate). After incubated at room temperature for 10-20 minutes, tubes are centrifugated for 20 min at 2,000-3,000 rpm. Collect the supernatant carefully as plasma samples. If precipitates appear during reservation, the sample should be centrifugated again.

  1. Urine samples

Collect urine into aseptic tubes. Collect the supernatant carefully after centrifuging for 20 min at 2,000-3,000 rpm. If precipitates appear during reservation, the sample should be centrifugated again. The preparation procedure of cerebrospinal fluid and pleuroperitoneal fluid is the same as that of urine sample.

  1. Cell samples

If you want to detect the secretions of cells, collect culture supernatant into aseptic tubes. Collect the supernatant carefully after centrifuging for 20 min at 2,000-3,000 rpm. If you want to detect intracellular components, dilute the cells to 1X100/ml with PBS (pH 7.2-7.4). The cells were destroyed to release intracellular components by repeated freezing and thawing. Collect the supernatant carefully after centrifuging for 20 min at 2,000-3,000 rpm. If precipitates appear during reservation, the sample should be centrifugated again.

  1. Tissue samples

Tissue samples are cut, weighed, frozen in liquid nitrogen and stored at -80℃ for future use. The tissue samples were homogenized after adding PBS (pH 7.4). Samples should be operated at 4. Collect the supernatant carefully after centrifuging for 20 min at 2,000-3,000 rpm. Aliquot the supernatant for ELISA assay and future use.

Notes:

  1. Sample extraction and ELISA assay should be performed as soon as possible after sample collection. The samples should be extracted according to the relevant literature. If ELISA assay can not be performed immediately, samples can be stored at -20℃.Repeated freeze-thaw cycles should be avoided.
  2. Our kits can not be used for samples with NaN3 which can inhibit the activity of HRP.

The quantitative detection of Human 2,3-DPG antigen is based on the ELISA (Enzyme-Linked Immunosorbent Assay) technique. The microplate wells are pre-coated with an antibody specific to Human 2,3-DPG. After incubation, the corresponding antigen in the sample or standards will bind to the immobilized antibody. After washing the microplate wells to remove all unbound sample material, horseradish peroxidase (HRP) labeled antibody conjugate is added, which binds to the antigen, forming antibody-antigen-enzyme-labeled antibody complex upon incubation. In a second wash step, all unbound material is removed. The immune complex formed by the bound conjugate is visualized by adding Tetramethylbenzidine (TMB) substrate, which gives a blue color reaction in wells containing the complex. A stop solution containing acid terminates the reaction, producing a yellow end-point coloration proportional to the amount of specific antigen present in the sample. Absorbance is measured at 450 nm using an ELISA microwell plate reader. Analysis is completed by comparing the OD values of samples to the standard curve.