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TAKARA MK301 Manual

Tracp & alp assay kit
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TRACP & ALP Assay Kit
I.
Description ................................................................................2
II. Introduction .............................................................................2
III. Kit components .......................................................................2
IV. Storage .......................................................................................3
V. Preparation of reagents .......................................................3
VI. Outline of Procedure ............................................................3
VII. Procedure .................................................................................4
VIII. Application examples ..........................................................5
IX. Reference ............................................................................... 10
X. Related Product ................................................................... 10
URL：http：//www.takara-bio.com
Table of Content
Cat. #MK301
v0804
1

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Summary of Contents for TAKARA MK301

  • Page 1: Table Of Contents

    Cat. #MK301 TRACP & ALP Assay Kit v0804 Table of Content Description ................2 II. Introduction ................2 III. Kit components ...............2 IV. Storage ..................3 V. Preparation of reagents ............3 VI. Outline of Procedure ............3 VII. Procedure .................4 VIII. Application examples ............5 IX.

  • Page 2: Description

    Cat. #MK301 TRACP & ALP Assay Kit v0804 I. Description TRACP&ALP Assay Kit allows for simultaneous detection of 2 enzymes which are in- volved in bone metabolism. TRACP which is an osteoclast enzyme marker and ALP an osteoblast enzyme marker. TRACP&ALP Assay Kit has been designed for simple and quick detection of ACP (Acid phosphatase) and ALP (Alkaline phosphatase) through the use of pNPP ( p -nitro-phenyl phosphate) substrate.

  • Page 3: Storage

    Cat. #MK301 TRACP & ALP Assay Kit v0804 4) Buffer for ACP 30 ml 0.5 M Sodium acetate, pH5.2 5) Buffer for ALP 30 ml 0.2 M Tris-HCl, pH9.5, 1 mM MgCl ＊ The concentration of Tris-HCl has changed for keeping pH stability from lot. 011.

  • Page 4: Procedure

    Cat. #MK301 TRACP & ALP Assay Kit v0804 VII. Procedure (1) For adherent cells cultured in a 96-well plate 1. Remove the culture supernatant with an aspirator, etc. 2. Add 200 μ l of physiological saline to each well, wash once, then discard the liquid.

  • Page 5: Application Examples

    Cat. #MK301 TRACP & ALP Assay Kit v0804 3. Dilute the cell lysate step-wise with physiological saline and add the prescribed amount (in the range of 5 - 50 μ l) to each well of the provided 96-well plate. Following this step, the procedure is the same as in (1) 4. - 5.
  • Page 6 Cat. #MK301 TRACP & ALP Assay Kit v0804 Plate row No. of cells 1 × 10 5 × 10 2.5 × 10 1.25 × 10 6.25 × 10 3.13 × 10 (cells/well) 5 μ l 2.275 2.271 1.732 0.920 0.581 0.329...
  • Page 7 Cat. #MK301 TRACP & ALP Assay Kit v0804 ＜ Results ＞ The bone marrow cell culture produces numbers of naturally differentiated osteo- clast-like TRACP-positive cells, and their TRACP activity could be measured using this kit. The table below shows the numbers of cells at the start of culturing and TRACP activity as absorbency at 405 nm.
  • Page 8 405 nm of the 96-well plate was measured using a plate reader. ＜ Result ＞ All alkaline phosphatase (BAP, CIAP, and SAP) from TAKARA BIO were available as a positive control for the kit. (5) Comparison of Tartrate-resistant Acid Phosphate (TRACP) and Alkaline Phospha- tase (ALP) by he preparation method of blood samples.
  • Page 9 Cat. #MK301 TRACP & ALP Assay Kit v0804 ＜ Results ＞ ＜ TRACP activity ＞ nm absorbanse TRACP citrate plasma（ PPP ） serum hemolysis serum ID No. x 20 x 40 x 80 x 20 x 40 x 80 x 20...

  • Page 10: Reference

    They shall not to be used products as food, cosmetics, or utensils, etc. Takara products may not be resold or transfered, modified for resale or transfer, or used to manufacture commercial products without written approval from TAKARA BIO INC.

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