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> Mouse Granzyme B (GZMB) ELISA Development Kit

Mouse Granzyme B (GZMB) ELISA Development Kit

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Cat# abx378329-15×96tests

Size : 15×96tests

Brand : Abbexa

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Mouse Granzyme B (GZMB) ELISA Development Kit

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Datasheet SDS
Mouse Granzyme B (GZMB) ELISA Development Kit for use in Sandwich ELISA assay development.

This ELISA Development Kit contains:
Component5 × 96 tests15 × 96 tests
Pretreated 96-well ELISA Plate515
Capture Antibody120 µl350 µl
Biotin-Conjugated Detection Antibody120 µl350 µl
HRP-Conjugate120 µl350 µl
Standard (2000 pg)1 vial15 vials

 

Please note that quantities and concentrations may change between different batches.

It is recommended to use this ELISA Development Kit with abx471002 ELISA Development Support Kit (Sandwich Method); alternatively, the following solutions can be prepared separately:

  • 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


Target Granzyme B (GZMB)
Reactivity Mouse
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 2000 pg/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 P04187 (UniProt, ExPASy)
Test Range 31.2 pg/ml - 2000 pg/ml
Detection Method Colormetric
Assay Type Sandwich
Sample Type Serum and plasma.
Sample Collection/Preparation Analyse immediately or store samples at 2-8°C (within 24 hrs). For long term storage, aliquot and store at -20°C or -80°C. Avoid multiple freeze-thaw cycles.
  • Serum: Samples should be collected into a serum separator tube. Coagulate the serum by leaving the tube undisturbed in a vertical position overnight at 4°C or at room temperature for up to 1 hr. Centrifuge at approximately 1000 × g for 20 mins. If precipitate appears, centrifuge again. Assay immediately or aliquot and store at -20°C or -80°C.
  • Plasma: Collect plasma using EDTA as an anticoagulant. Centrifuge for 15 mins at 1000 × g, within 30 mins of collection. If precipitate appears, centrifuge again. Avoid hemolytic samples.
Availability Shipped within 5-10 working days.
Note THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
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 Granzyme B (GZMB)