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Merck
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387A

Sigma-Aldrich

Leukocyte Acid Phosphatase (TRAP) Kit

Kit formulated with all liquid reagents

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About This Item

UNSPSC 코드:
12352106
eCl@ss:
42010102
NACRES:
NA.47
387A 제품은 고객 맞춤형 제품으로 웹사이트에서 판매할 수 없습니다. 고객지원팀으로 연락바랍니다.

유통기한

Expiry date on the label.

IVD

for in vitro diagnostic use

dilution

(for histology)

응용 분야

hematology
histology

배송 상태

wet ice

저장 온도

2-8°C

관련 카테고리

원리

Peripheral blood or bone marrow preparations are fixed to a microscope slide. The resulting film is incubated in a solution of Naphthol AS-BI phosphoric acid and freshly diazotized Fast Garnet GBC. Kit is used to demonstrate acid phosphatase and tartrate resistant acid phosphatase (TRAP) in blood, bone marrow and tissue touch preparations.

키트 구성품 전용

제품 번호
설명
  • Hematoxylin Solution, Gill No. 3 (kit only) 50 mL

키트 구성품 역시 별도로 이용 가능함

제품 번호
설명
SDS
  • 3863Acetate Solution, 2.5 M, pH 5.2 50 mLSDS
  • 915Citrate Solution, pH 3.6±0.1 (25 °C), 27 mM 50 mLSDS
  • 3872Fast Garnet GBC Base Solution 10 mLSDS
  • 3871Naphthol AS-BI Phosphoric Acid Solution 10 mLSDS
  • 914Sodium nitrite solution 10 mLSDS
  • 3873Tartrate Solution 10 mLSDS

픽토그램

Health hazardCorrosionExclamation mark
GHS08,GHS05,GHS07

신호어

Danger

유해 및 위험 성명서

H290,H302,H314,H350,H373

Hazard Classifications

Acute Tox. 4 Oral - Carc. 1B - Eye Dam. 1 - Met. Corr. 1 - Skin Corr. 1B - STOT RE 2 Oral

표적 기관

Kidney

Storage Class Code

6.1C - Combustible acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects

WGK

WGK 3

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시험 성적서(COA)

Lot/Batch Number
0000378672
0000362640
0000362640
0000378672
0000293208

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COA 검색

이 제품을 이미 가지고 계십니까?

문서 라이브러리에서 최근에 구매한 제품에 대한 문서를 찾아보세요.

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L Wei et al.
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Yun Zhang et al.
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Yu Wang et al.
Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 47(6), 2291-2306 (2018-07-06)
Osteoporosis is a commonly occurring condition marked by a loss of bone density. Previous evidence has highlighted the roles played by microRNAs as potential treatment tools for the disease. At present, the influence of long non-coding RNAs (lncRNAs) on the
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질문

1–10 / 14 질문  

  1. Is there a staining protocol available for culturing cells using either the 386A or 387A kits for blood smears?

    1 답변

    1. The 387A kit does not have a published protocol for staining cultured cells. The kit is approved for human In Vitro Diagnostic Use and was primarily intended for detecting TRAP positive cells in peripheral blood or bone marrow, particularly for the detection of TRAP positive cells associated with Hairy Cell Leukemia. However, the procedure included with the kit can be used.

      First, remove the media, rinse briefly in isotonic PBS or suitable cell culture media, and then air dry the cells. Once dried, the cells can be fixed onto chamber slides, microtiter plates, or culture flasks. After fixing, the cells are washed.

      The procedure mentions staining slides for Beaker A and Beaker B. When staining cultured cells, think of staining duplicate wells.
      One set of cells will be incubated in the presence of tartrate (Beaker B), and the second set will be incubated without tartrate (Beaker A), which serves as the positive control.

      Staining cells without tartrate is essential for ensuring the kit is performing to expectations. The kit was designed to stain an equal number of slides/wells with and without tartrate, and there is a separate product number (387-3) for the tartrate reagent, which may be ordered separately.

      The kit instructions call for preparing a specific volume of staining reagent for Beaker A and Beaker B. If needed, the volume of reagent can be scaled down, but once prepared, the staining solution is for one-time use only and cannot be stored for later use.

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  2. Why does the kit procedure call for staining duplicate slides Beaker A and Beaker B?

    1 답변

    1. Beaker A stains all isoforms of Acid Phosphatase, while Beaker B stains only the Tartrate Resistant Acid Phosphatase (TRAP) isoenzyme of Acid Phosphatase. Beaker A serves as a control and it is recommended to run both Beaker A and Beaker B. Running only Beaker B makes it impossible to determine if the kit is working properly, whether the enzyme is present, or if the procedure was performed properly.

      If staining is present with slides for Beaker B, staining should also be present for the slides stained in Beaker A. However, if only Beaker B is utilized and no staining is present, the problem could be with the kit, the specimen, or how the procedure was performed. Therefore, Beaker A should always be run.

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  3. Can a paraffin section be utilized as a positive control for the TRAP 386A or 387A procedures?

    1 답변

    1. Paraffin sections are not ideal controls for use with either the 386A or 387A TRAP kits. When sections are cut from a bone, the bone must first be fixed, then decalcified in EDTA, and finally, paraffin embedded. While the Tartrate Resistant Acid Phosphatase (TRAP) enzyme is relatively hardy, it may not always survive decalcification and paraffin processing. The most effective method of decalcification involves removing calcium with EDTA solutions. Harsh acids such as HCL, Sulfuric Acid, and Formic Acid typically deactivate the TRAP enzyme, isoenzyme 5.

      Suitable controls recommended for use with the kit are either peripheral blood smears or buccal smears. It is not known if buccal smears from animals are TRAP positive.
      With a peripheral blood smear, Beaker A slides should be positive, and Beaker B slides should be negative.
      With a buccal smear, both Beaker A and Beaker B slides should be positive.
      If peripheral blood smears or buccal smears are negative for both Beaker A or Beaker B, there may be a problem with the kit or possibly with the staining technique.

      In such cases, it is advisable to contact Technical Service for troubleshooting or to file a complaint.

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  4. I will perform TRAP staining to detect osteoclasts. Which mixture should I use for this, beaker A or beaker B?

    1 답변

    1. When staining osteoclasts, both Beaker A and Beaker B are recommended. Beaker A will stain all 7 different isoenzymes of Acid Phosphatase. No tartrate is added to the staining solution for Beaker A. Beaker B includes tartrate. The only cells that will stain in the presence of tartrate are the hairy cells of hairy cell leukemia and differentiated osteoclasts (isoenzyme 5). The remaining cells are inhibited by the presence of tartrate and will not display positive staining.
      Please see the link below to review the protocol for this kit:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/539/315/387.pdf

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  5. Is the Leukocyte Acid Phosphatase TRAP kit compatible with paraffin sections that have been decalcified?

    1 답변

    1. The 386A and 387A kits are approved for In Vitro Diagnostic Use on peripheral blood smears and bone marrow smears. However, there are no claims for these kits working on paraffin sections, and they are not intended or approved for use with human bone.

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  6. Can the TRAP enzyme still be demonstrated after fixing mouse brain and soft tissue in 10% formalin and decalcifying in 10% EDTA for 4 weeks? Or was the length of time spent in 10% EDTA excessive, resulting in the deactivation of the TRAP enzyme?

    1 답변

    1. It is uncertain whether 4 weeks in 10% EDTA is too much time for decalcification. The most accurate methods for checking whether decalcification is complete typically involve using an X-ray or a chemical endpoint determination for the removal of calcium.

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  7. There are two kits for Acid Phosphatase.  What are the major differences between the two kits?

    1 답변

    1. Both kits stain for Leukocyte Acid Phosphatase. The 386A kit was developed first. Some of the reagents are packaged in gelatin capsules. The capsules must be broken open to dissolve the reagents. The gelatin capsules will not dissolve in the staining solution in any reasonable period of time. Once dissolved the staining reagent should be viewed as unstable and the slides should be stained as soon as the temperature reaches 37°C. The 386A kit uses a Citrate Acetone fixative. The 387A kit, however, is formulated with all liquid reagents; there are no gelatin capsules. As a result the 387A kit is considered more user friendly. Once the reagents are prepared as outlined in the package insert, the working solution is again considered unstable and the slides should be stained as soon as the temperature reaches 37°C. The pricing of the kit is slightly less, but the expiration dating may also be slightly less. With either kit, it is recommended that the customer check expiration dating before ordering. The 387A kit uses a Citrate Acetone Formaldehyde fixative. Labs wishing to avoid use of formaldehyde should consider the 386A kit.

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  8. Can Product 387A, Acid Phosphatase, Leukocyte (TRAP) Kit, be used to stain cultured cells?

    1 답변

    1. Yes.  The product insert contains information regarding only the approved in vitro diagnostic use of the product. Researchers can and do use the kit to stain cultured cells.  The kit is most commonly used to determine when various cell types differentiate into osteoclasts.

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  9. I am staining cultured cells with Product 387A, Acid Phosphatase, Leukocyte (TRAP) Kit.  Why are the cells incubated in Beaker A and Beaker B both positive?

    1 답변

    1. Cells in Beaker A will demonstrate all isoforms of Acid Phosphatase.  Cells in Beaker B will demonstrate only Tartrate Resistant Acid Phosphatase (TRAP) cells, i.e. those cells which have already differentiated into osteoclasts.

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  10. How can I easily prepare a TRAP positive control for use with Product 387A, Acid Phosphatase, Leukocyte (TRAP) Kit?

    1 답변

    1. A buccal smear from the inside of the human cheek makes an excellent positive control.  The epithelioid cells are TRAP positive.

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