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PMC-AK04F-COS 破骨細胞染色試劑盒 TRAP Staining Kit英文說明書

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PMC-AK04F-COS 破骨細胞染色試劑盒 TRAP Staining Kit英文說明書

破骨細胞染色試劑盒/抗酒石酸酸性磷酸酶（TRAP）染色試劑盒

酸性磷酸酶(AcidPhosphatase)，也稱酸性磷酸酯酶，在酸性條件下可以催化磷酸酯鍵的水解。酸性磷酸酶是一個蛋白家族，哺乳動物中其分子量 從18kD到100kD不等。酸性磷酸酶主要分為兩類，一類為抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase, TRAP)，一類為抗氟離子酸性磷酸酶。

抗酒石酸酸性磷酸酶是一種糖基化的含金屬蛋白酶，在破骨細胞(osteoclast)和破軟骨細胞(chondroclast)中高表達。在活化的巨 噬細胞和神經(jīng)元中也有表達。抗酒石酸酸性磷酸酶在細胞信號轉(zhuǎn)導、細胞增殖和分化等方面起重要作用。

  • 產(chǎn)品名稱:TRAP Staining Kit
  • 保存溫度:4C DNF
  • 運輸溫度:（效期最長6個月）

PMC-AK04F-COS 破骨細胞染色試劑盒 TRAP Staining Kit英文說明書

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

Component Quantity Storage
Tartrate-containing?Buffer 50ml 4°C
Chromogenic?Substrate 10?Vials 4°C

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.