GBL® ROSA-STAIN

5057-Alkaline Phosphatase Leukocyte Staining Kit

Title Description
REF No 5057
Purpose Light microscopic examination of alkaline phosphatase activity in blood, bone marrow or tissue samples
used.
Ingredients A) Citrate solution pH: 5.4: Sodium chloride, Citric acid, Trisodium citrate.
B) FRV red dye: Fast Red Violet LB Salt.
C) HCl solution: Hydrochloric acid.
D) Sodium nitrite solution: Sodium nitrite.
E) Naphthol AS-BI phosphate: Naphthol AS-BI phosphate.
F) AMPD solution: AMPD.
G) Hematoxylin Solution Gill-3: Hematoxylin, Aluminum sulfate, Sodium iodate, Glacial acetic acid.
Working Principle Alkaline phosphatase leukocyte stain is a staining method used to identify leukocytes (white blood cells). This method targets the enzyme alkaline phosphatase, which is found in the cell membranes of leukocytes. Alkaline phosphatase is an enzyme found on the outer surface of the cell membrane and is abundant in leukocytes. Alkaline phosphatase enzyme hydrolyzes the phosphate molecules in the staining solution. As a result of this reaction, phosphate residues are formed on the cell membranes of the cells. Under a microscope, the cell membranes of leukocytes are stained to make sure that the stain shows alkaline phosphatase activity. This makes leukocytes easily distinguishable from other blood cells.
Warnings, Precautions, Recommendations Caution. Harmful if swallowed, in contact with skin or inhaled. May cause damage to organs through prolonged or repeated exposure. Suspected of causing cancer. Rinse mouth. May cause respiratory irritation. Avoid inhalation of dust/fume/gas/mist/vapor/spray. Avoid inhalation of dust/fume/gas/mist/mist/vapor/spray. Wear protective gloves/protective clothing/eye protection/face protection. If you feel unwell, call the POISON CENTER or doctor/physician.
IF INGESTED: If you feel unwell, call the NATIONAL POISON ADVICE CENTER, TELEPHONE 114 or a doctor/physician. IN CASE OF CONTACT WITH EYES: Rinse carefully with water for several minutes. Remove contact lenses if fitted and easy to do so. Continue rinsing.
Application 1. Preparation of FRV Alkaline Solution:
Add 1 mL of HCl solution to 5 mg of FRV dye. Dissolve by stirring.
2. Preparation of Naphtol AS-BI phosphate solution:
Add 1 mL of AMPD solution to a tube containing Naphtol AS-BI. Dissolve by stirring.
3. Preparation of incubation solution:
Mix 1 mL of sodium nitrite solution (Component C) and 1 mL of FRV Alkaline solution. Wait for 2 minutes.
Add 45 mL of distilled water. (at 18-26 C)
Add 1 mL of Naphtol AS-BI phosphate solution. Mix well and pour into the jar.
4. Preparation of fixative solution:
Mix 13 mL of ‘’A-Citrate solution’’, 33 mL of acetone and 4 mL of 37% formaldehyde solution.
5. Immerse the preparation in the “fixative solution” container brought to room temperature. Wait 30 seconds. Wash gently with distilled water for 45 seconds. Do not allow the preparation to dry.
6. Immerse the preparation in the “incubation solution” container. Incubate at 15-25 °C for 15 minutes. Protect your immersed preparations from sunlight. Do not reuse this solution. Dispose of them.
7. After 15 minutes of incubation, remove your preparations from the jar and wash them with distilled water for 2 minutes. Do not allow the preparations to dry.
8. Stain the preparation with E- Hematoxylin solution Gill-3 for 2 minutes.
9. Wash with tap water and dry.
10. Examine under a microscope.
Theoretical Results
Alkaline phosphatase activity Red

Related Media
Shelf Life 1 year
Shelf Life After Opening the Lid
Storage Temperature 2-8 °C
pH (at 25°C) A) Citrate solution pH: 5.2
F) AMPD solution pH: 9.5
Transportation Information
Ordering and Packaging Information
Order No Unit Volume Packaging Type Quantity in a Box
445057050010 Piece 50 mL 50 ml, White Bottle with HDPE Dropper 1 kit

Keywords Alkali Phosphatase Leukocyte, Phosphatase Leukocyte stain, Alkaline phosphatase activity
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