PMC-AK04F-COS 破骨细胞染色试剂盒 TRAP Staining Kit英文说明书

破骨细胞染色试剂盒/抗酒石酸酸性磷酸酶（TRAP）染色试剂盒

酸性磷酸酶(AcidPhosphatase)，也称酸性磷酸酯酶，在酸性条件下可以催化磷酸酯键的水解。酸性磷酸酶是一个蛋白家族，哺乳动物中其分子量 从18kD到100kD不等。酸性磷酸酶主要分为两类，一类为抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase, TRAP)，一类为抗氟离子酸性磷酸酶。

抗酒石酸酸性磷酸酶是一种糖基化的含金属蛋白酶，在破骨细胞(osteoclast)和破软骨细胞(chondroclast)中高表达。在活化的巨 噬细胞和神经元中也有表达。抗酒石酸酸性磷酸酶在细胞信号转导、细胞增殖和分化等方面起重要作用。

• 产品名称:TRAP Staining Kit
• 保存温度:4C DNF
• 运输温度:（效期最长6个月）

PMC-AK04F-COS 破骨细胞染色试剂盒 TRAP Staining Kit英文说明书

TRAP Staining Kit

Cat. No. PMC-AK04-COS

The TRAP Staining Kit (Cat.No.PMC-AK04F-COS) is used for the staining of Tartrate-Resistant Acid Phosphatase in osteoclasts. Bone mass is controlled by the balance between the activity of osteoblasts bone formation and the activity of osteoclasts. Alkaline phosphatase (ALP: ALP staining kit, Cat.No.PMC-AK20-COS) and tartrate-resistant acid phosphate are used as markers for osteoblasts and osteoclasts, respectively.

【 I 】Background

【 II 】Kit Components

One kit contains regents for 10 × 96-well plate

【 II 】Additional Materials Required

One kit contains regents for 10 × 96-well plates

・10% Formalin Neutral Buffer Solution

・Phosphate Buffered Saline (PBS)

・Distilled or deionized water

【 II 】Protocol

1 、Remove culture medium. Wash each well once with 100uL of PBS.

2、Add 50uL of the 10% Formalin Neutral Buffer Solution to each well and fix for 5 minutes at room temperature

3、Wash each well with 250uL of deionized water. (× 3 times)

4、Dissolve 1 vial of Chromogenic Substrate with 5mL of Tartrate-containing Buffer

5 、Add 50uL of Chromogenic Substrates to each well.

6、Incubate at 37°C for 20-6、60 minutes. Adjust incubation time until stained TRAP is clearly showing the result in figure 1.

7    Wash with deionized water to stop the reaction.

Add 50uL of the 10% Formalin Neutral Buffer Solution to each well and fix for 5 minutes at room temperature

For research use only, Not for diagnostic use. Please read this manual thoroughly before use.

【 VI 】References

• BEN A. A. SCHEVEN, JONE S. MILNE, SIMON P. ROBINS. A SEQUENTIAL CULTURE APPROACH TO STUDY OSTEOCLAST DIFFERENTIATIONFROM NONADHERENT PORCINE BONE MARROW CELLS. In Vitro Cell. Dev. Biol. July-August (1998). Animal 34: 568-577.
• Martha Somerman, Janice E. Berry, Zhila Khalkhali-Ellis, Philip Osdoby, Robert U. Simpson. Enhanced Expression of α v Integrin Subunit and Osteopontin during Differentiation of HL-60 Cells along Monocytic Pathway. EXPERIMENTAL CELL RESEARCH 216, 335-341(1995).
• Ichiro Itonaga, Afsie Sabokbar, Susan D. Neale, Nicholas A. Athanasou. 1,25-Dihydroxyvitamin D3 and Prostaglandin E2 Act Directly on Circulating Human Osteoclast Precursors. Biochemical and Biophysical Research Communications 264, 590-595(1999).
• Hiroshi Takayanagi, Kouetsu Ogasawara, Shigeaki Hida, Tomoki Chiba, Shigeo Murata, Kojiro Sato, Akinori Takaoka, Taeko Yokochi, Hiromi Oda, Keiji Tanaka, Kozo Nakamura and Tadatsugu Taniguchi. T-cell-mediated regulation of osteoclastogenesis by signaling cross-talk between RANKL and IFN-α Nature, Vol. 408, No.6812, 600-605, 30 November 2000.
• Morinobu,, Biao, W., Tanaka, S., Horiuchi, M., Jun, L., Tsuji,G., Sakai, Y., Kurosaka, M., Kumagai, S. (-)-Epigallocatechin-3-Gallate Suppresses

Osteoclast Differentiation and Ameliorates Experimental Arthritisv in Mice. Arthritis Rheum. 58, 2012-2018 (2008)

• Hase,, Kanno, Y., Kojima, H., Sakurai, D., Kobata, T. Coculture of Osteoclast Precursors With Rheumatoid Synovial Fibroblasts Induces Osteoclastogenesis via Transforming Growth Factor beta-Mediated Down-Regulation of Osteoprotegerin. Arthritis Rheum. 58, 3356-3365 (2008)
• Cao,, Kuboyama, N. A Biodegradable Porous Composite Scaffold of PGA/beta-TCP for Bone Tissue Engineering. Bone. 46, 386-395 (2010)
• Okada,, Hamada, N., Kim, Y., Takahashi, Y., Sasaguri, K., Ozono, S., Sato, S. Blockade of Sympathetic beta-receptors Inhibits Porphyromonas Gingivalis-induced Alveolar Bone Loss in an Experimental Rat Periodontitis Model. Arch. Oral Biol. 55, 502-508 (2010)
• Okamoto,, Ohnishi, T., Bandow, K., Kakimoto, K., Chiba, N., Maeda, A., Fukunaga, T., Miyawaki, S., Matsuguchi, T. Reduction of Orthodontic Tooth Movement by Experimentally Induced Periodontal Inflammation in Mice. Eur. J. Oral Sci. 117, 238-247 (2009)

【 VII 】Notice to Purchaser
This product is to be used for Research Purposes Only. It is not to be used for Drug or Diagnostic Purposes, nor is it intended for Human Use. Primary cell products may not be resold, modified for resale, or used to manufacture commercial products without the express written consent of Primary Cell Co., Ltd.

Except as otherwise expressly set forth in this use manual, Primary Cell dose not make any representation or warranties or conditions of any kind, either express or implied, with respect to the products, or information disclosed hereunder, including, but not limited to, the implied warranties of merchantability, fit for a particular purpose, or noninfringement of the intellectual property rights of third parties.