Acute Tox. 4 Oral - Carc. 1B - Eye Dam. 1 - Met. Corr. 1 - Skin Corr. 1A - STOT RE 2 Oral - STOT SE 3
Kidney, Respiratory system
6.1C - Combustible, acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects
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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.
The kit results are qualitative. A microscope is used to determine the presence or absence of staining within the cytoplasm of the cell.
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.
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.
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.
A buccal smear from the inside of the human cheek makes an excellent positive control. The epithelioid cells are TRAP positive.
A light microscope is used to view the staining within the cytoplasm of the cells. If staining in a microtiter plate, it should be possible to use an inverted microscope to view the staining, although this has not been confirmed at Sigma-Aldrich.
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