Human Beta-2-Microglobulin (B2M) ELISA Development Kit

A comprehensive kit for developing sandwich ELISA assays to detect Beta-2-Microglobulin (B2M). The kit contains all necessary reagents for 5 or 15 x 96 tests.

Kit Components:

– Pretreated 96-well ELISA Plate
– Capture Antibody
– Biotin-Conjugated Detection Antibody
– HRP-Conjugate
– Standard (20,000 ng)

Recommended Accessories:

BTA14596  ELISA Development Support Kit (Sandwich Method)

Buffer Preparation:

For optimal results, prepare the following buffers:

– Coating Buffer: 1X Citrate-Buffered Saline
– Blocking Buffer: 1X PBS containing 0.5-3% BSA
– Wash Buffer: 3% Tris
– Standard and Sample Diluent Buffer: 1X PBS containing 0.5-3% BSA
– Antibody and HRP-Conjugate Diluent Buffer: 1X PBS containing 0.5-3% BSA
– Stop Solution: 5% Sulfuric Acid

Cat. No: 14596

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Description

Target Beta-2-Microglobulin (B2M)
Reactivity Human
Tested Applications ELISA
Recommended dilutions Capture Antibody: 1/500 – 1/1000, Biotin-conjugated Detection Antibody: 1/500 – 1/1000, HRP-Conjugate: 1/500 – 1/1000. Optimal dilutions/concentrations should be determined by the end user.
Reconstitution Reconstitute the standard with 1 ml of Standard Diluent to obtain a stock standard solution of 20000 ng/ml. Further dilute by a factor of 10 to give the highest standard, 2000 ng/ml. Label tubes in preparation for the serial dilutions (1000, 500, 250, 125, 62.5, and 31.25 ng/ml). Aliquot 0.5 ml of the Standard Diluent into each tube. Add 0.5 ml of the highest standard solution into the 1st tube and mix thoroughly. Transfer 0.5 ml from the 1st to 2nd tube, mix thoroughly, and so on.
Storage Aliquot and store at -20°C in the dark. Avoid repeated freeze/thaw cycles.
UniProt Primary AC P61769 (UniProt, ExPASy)
Test Range 31.25 ng/ml – 2000 ng/ml
Detection Method Colormetric
Assay Type Sandwich
Sample Type Serum and plasma.
Availability Shipped within 5-10 working days.
Note This product is for research use only.
Directions for use Bring all components to room temperature (18-25°C) and briefly spin or centrifuge the vials before use. Working solutions should be prepared and used immediately.
Recommended Procedure:

  1. Dilute the Capture Antibody to working concentration using Coating Buffer. Immediately coat the 96-well plate with diluted Capture Antibody (100 µl per well). Seal the plate and incubate at 2-8 °C overnight.
  2. Remove the liquid from each well. Do not wash. Block the plate with Blocking Buffer (200 µl per well) at 37 °C for 1 hour.
  3. Remove the liquid from each well. Do not wash. Either proceed with the following steps immediately or dry the plate at 37 °C for 30 minutes, then store at -20 °C with dessicant for up to 6 months.
  4. Add 100 µl of standards or sample into the appropriate wells. Cover with a plate sealer and incubate at 37 °C for 1.5 hours.
  5. Remove the liquid from each well. Do not wash. Add appropriately diluted Biotin-Conjugated Detection Antibody (100 µl per well). Cover the plate with a new plate sealer and incubate at 37 °C for 1 hour.
  6. Remove the liquid from each well. Wash with Wash Buffer (350 µl per well) and allow to soak for 1-2 min. Remove the liquid by inverting and tapping the plate on to absorbent paper. Repeat the wash process 3 times.
  7. Add appropriately diluted HRP-Conjugate (100 µl per well). Cover the plate with a new plate sealer and incubate at 37 °C for 30 min.
  8. Repeat the wash process in Step 6, for a total of 5 times.
  9. Add Substrate Solution (90 µl per well). Cover the plate with a new plate sealer and incubate at 37 °C for 15-30 min. Keep the plate in the dark and avoid exposure to light.
  10. Add Stop Solution (50 µl per well). Tap the side of the plate to ensure thorough mixing.
  11. Measure the absorbance immediately using a microplate reader set at 450 nm.

Research Articles on Beta-2-Microglobulin (B2M)

Data sheet

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SDS

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