| Atribut | pojedinosti |
|---|---|
| Mačka | G1480 |
| Veličina | 4 x 2mL, 4 x 10mL, 4 x 20mL |
| Skladištenje | -20°C, avoid light, valid for 6 mjeseca |
Kit Components:
| Reagens | 4x2mL | 4x10mL | 4x20mL | Skladištenje |
|---|---|---|---|---|
| Reagent A: ALP Fixative | 2ml | 10ml | 20ml | Rt, avoid light |
| Reagent B: ALP Incubation Solution | B1: AS-BI Solution 1mL | 5ml | 10ml | -20°C, avoid light |
| B2: FBB Solution 1mL | 5ml | 10ml | -20°C, avoid light | |
| Reagent C: Nuclear Fast Red Solution | 2ml | 10ml | 20ml | 2-8°C, avoid light |
| Reagent D: Methyl Green Solution | 2ml | 10ml | 20ml | Rt, avoid light |
Uvod:
Alkaline phosphatase (ALP or AKP) is widely distributed in mammalian tissues, mainly at active substance exchange sites like cell membranes, endoplasmic reticulum, Golgi complex, lysosomes, neutrophil granules, and smooth muscle cell membranes. The ALP Staining Kit (Kaplow Method) utilizes an azo coupling method to demonstrate ALP activity, providing reliable results compared to metal salt precipitation methods.
Self-Provided Materials:
Glass slide, Wet box, Optical microscope
Protocol (for reference only):
For Smear or Section Stain:
- Fix smears or sections in ALP Fixative for 3min, then wash with distilled water.
- Apply ALP Incubation Solution, incubate in a wet box in the dark for 15-20min, then wash with distilled water.
- Re-dye with Nuclear Fast Red Solution or Methyl Green Solution for 3-5min.
- View sections under a microscope after washing or sealing with glycerol gelatin.
For Adherent Cells Stain:
- Rinse cultured cells with PBS, then fix them with ALP Fixative or 4% PFA, followed by washing with PBS.
- Apply ALP Incubation Solution, incubate in a wet box in the dark for 15-20min, then wash with PBS.
- Re-dye with Nuclear Fast Red Solution or Methyl Green Solution for 3-5min.
- Wash with PBS and view sections under a microscope.
Proizlaziti:
| Proizlaziti | Color |
|---|---|
| Positive site of ALP | Blue |
| Nucleus | Red or Green |
Criteria for Blood and Bone Marrow Smears Judging:
| Positive Level | Dyeing Characteristics |
|---|---|
| 0 | No Granule |
| 1 | Slightly Granule |
| 2 | Medium-Grade Granule |
| 3 | Majority Granule |
| 4 | Full Granule |
Bilješka:
- Use fresh samples and fix sections promptly to preserve enzyme activity.
- Handle cultured cells gently during staining to avoid damage or loss.
- ALP Incubation Solution may lose effectiveness quickly, use promptly.
- Re-dye with Nuclear Fast Red Solution or Methyl Green Solution.
- Include a positive control for each sample.
Referenca:
Yu Jiang, Dantian Zhu, Wenfeng Liu, et al. Hedgehog pathway inhibition causes primary follicle atresia and decreases female germLine stem cell proliferation capacity or stemness. Stem Cell Research & Therapy. July 2019. (IF 4.627)
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