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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 Code:
12352106
eCl@ss:
42010102
NACRES:
NA.47
Product 387A is not currently sold in your country. Contact Technical Service

Quality Level

500

shelf life

Expiry date on the label.

IVD

for in vitro diagnostic use

application(s)

hematology
histology

shipped in

wet ice

storage temp.

2-8°C

Related Categories

Principle

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.

Kit Components Only

Product No.
Description
  • Hematoxylin Solution, Gill No. 3 (kit only) 50 mL

Kit Components Also Available Separately

Product No.
Description
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

Signal Word

Danger

Hazard Statements

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

Target Organs

Kidney

Storage Class Code

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

WGK

WGK 3

Choose from one of the most recent versions:

Certificates of Analysis (COA)

Lot/Batch Number
0000378672
0000362640
0000362640
0000378672
0000293208

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Nan Shang et al.
Nutrients, 11(9) (2019-09-25)
Ovotransferrin, a member of the transferrin family, is the second main protein found in egg white. Ovotransferrin was reported to have antimicrobial, antioxidant, and immunomodulating activities. The aim of this work was to characterize the cellular and molecular functions of
Huey-En Tzeng et al.
Scientific reports, 8(1), 11190-11190 (2018-07-27)
NC-8 (ent-16-oxobeyeran-19-N-methylureido) is an isosteviol-derived analogue with multiple biological effects, including anti-inflammation and anti-bacterial activities and inhibition of HBV viral surface antigen gene expression. In this study, we explored the effects of NC-8 on the formation of osteoclasts from RAW
Jihai Wang et al.
Frontiers in pharmacology, 9, 64-64 (2018-02-24)
Lipopolysaccharide (LPS) can induce bone loss by stimulating bone resorption. Natural compounds have great potential for the treatment of osteolytic bone diseases. Magnesium lithospermate B (MLB) plays an important role in protecting against oxidative damage and also has potential anti-inflammatory
L Wei et al.
Osteoporosis international : a journal established as result of cooperation between the European Foundation for Osteoporosis and the National Osteoporosis Foundation of the USA, 25(8), 2089-2096 (2014-05-09)
Recently, the use of the pharmacological agent strontium ranelate has come to prominence for the treatment of osteoporosis. While much investigation is focused on preventing disease progression, here we fabricate strontium-containing scaffolds and show that they enhance bone defect healing
Yun Zhang et al.
Biological & pharmaceutical bulletin, 39(12), 2028-2035 (2016-12-03)
Osteolysis induced by chronic Gram-negative bacterial infection underlies many bone diseases such as osteomyelitis, septic arthritis, and periodontitis. Drugs that inhibit lipopolysaccharide (LPS)-induced osteolysis are critically needed for the prevention of bone destruction in infective bone diseases. In this study
View All Related Papers

Questions

1–10 of 14 Questions  

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

    1 answer

    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.

      Helpful?

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

    1 answer

    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.

      Helpful?

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

    1 answer

    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.

      Helpful?

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

    1 answer

    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

      Helpful?

  5. 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 answer

    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.

      Helpful?

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

    1 answer

    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.

      Helpful?

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

    1 answer

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

      Helpful?

  8. Are results from Product 387A, Acid Phosphatase, Leukocyte (TRAP) Kit, qualitative or quantitative?

    1 answer

    1. The kit results are qualitative.  A microscope is used to determine the presence or absence of staining within the cytoplasm of the cell.

      Helpful?

  9. Are paraffin-embedded tissue sections suitable for use with Product 387A, Acid Phosphatase, Leukocyte (TRAP) Kit?

    1 answer

    1. The kit has been shown to work well with blood smears and cultured cells.  Whether the kit will work on paraffin sections depends upon the size of the bone, the decalcification (decal) agent used, and the paraffin processing schedule utilized.  In general the smaller the bone fragment and the more mild the decal, the more likely the kit would work. EDTA for short periods of time seems to be the only acceptable decal agent.  As the bone sections become larger and the time in the decal becomes longer, the less likely it becomes that the kit will provide positive results.

      Helpful?

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

    1 answer

    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.

      Helpful?

1–10 of 14 Questions  

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