| Description | Annexins are a family of calcium-dependent phospholipid-binding proteins that preferentially bind phosphatidylserine (PS). Under normal physiologic conditions, PS is predominantly located in the inner leaflet of the plasma membrane. Upon initiation of apoptosis, PS loses its asymmetric Annexins are a family of calcium-dependent phospholipid-binding proteins that preferentially bind phosphatidylserine (PS). Under normal physiologic conditions, PS is predominantly located in the inner leaflet of the plasma membrane. Upon initiation of apoptosis, PS loses its asymmetric distribution across the phospholipid bilayer and is translocated to the extracellular membrane leaflet marking cells as targets of phagocytosis. Once on the outer surface of the membrane, PS can be detected by fluorescently labeled Annexin V in a calcium-dependent manner. In early-stage apoptosis, the plasma membrane excludes viability dyes such as propidium iodide (PI), 7-AAD. These cells will stain with Annexin V but not a viability dye, thus distinguishing cells in early apoptosis. However, in late stage apoptosis, the cell membrane loses integrity thereby allowing Annexin V to also access PS in the interior of the cell. A viability dye can be used to resolve these late-stage apoptotic and necrotic cells (Annexin V, viability dye-positive) from the early-stage apoptotic cells (Annexin V positive, viability dye-negative). This kit is suitable for the identification and enumeration of dead cells, such as apoptotic or necrotic cells, by flow cytometry. A1456530Components20T50T100TStorageQuantity Per TestA1456530A10X Annexin V Binding Buffer5 mL10 mL20 mL2-8℃200 µL per 0.5-1.0x10⁵ cellsA1456530BAnnexin V-FITC40 µL100 µL200 µL2-8℃. Store in the dark.2 µL per 0.5-1.0x10⁵ cellsA1456530C7-AAD Staining Solution 100 µL250 µL500 µL2-8℃. Store in the dark.5 µL per 0.5-1.0x10⁵ cellsNote: The recommended number of cells to stain per test is 0.5-1.0x10⁵ cells.Precautions 1. Please try to avoid light when using to slow down the quenching of fluorescence. 2. 7-AAD Solution is toxigenic and mutagenic; handle with care. 3. Since the binding of annexin V to phosphatidylserine (PS) is calcium-dependent, it is critical to avoid buffers containing EDTA or other calcium chelators during Annexin V experiments. Instruction for use 1. Dilute 10x Binding Buffer to 1x using distilled water (1 mL 10x Binding Buffer + 9 mL ddH2O). 2. Wash cells twice with cold PBS and then resuspend the desired amount of cells in Annexin V Binding Buffer at a concentration of 0.5-1.0x10⁶ cells /mL . 3. Add 2 µL of FITC Annexin V and 5 µL 7-AAD to 100 µL of the cell suspension. 4. Add 100 µL of 1x Binding Buffer to each assay. Gently vortex the cells and incubate for 10 min at RT (25°C) in the dark. 5. Analyze by flow cytometry within 1 hr... Read More | Alanine Aminotransferase (ALT), also known as serum glutamic-pyruvic transaminase (SGPT), is a pyridoxal-phosphate-dependent enzyme that catalyzes the reversible transfer of an amino group from alanine to α-ketoglutarate, generating pyruvate and glutamate. ALT is found primarily in liver and Alanine Aminotransferase (ALT), also known as serum glutamic-pyruvic transaminase (SGPT), is a pyridoxal-phosphate-dependent enzyme that catalyzes the reversible transfer of an amino group from alanine to α-ketoglutarate, generating pyruvate and glutamate. ALT is found primarily in liver and serum, but occurs in other tissues as well. Hepatocellular injury often results in an increase of serum ALT levels and serum ALT levels can be used as a marker for liver injury.ALT Activity Assay kit has been used to determine the activity of alanine aminotransferase (ALT) in serum samples... Read More | Inquire | Inquire | This plant protein extraction kit can extract soluble plant proteins from fresh, frozen, or dried plant tissues. Suitable for protein extraction from various plants and different parts of plants (such as roots, stems, leaves, flowers, seeds, etc.), with high extraction efficiency, high protein yieldThis plant protein extraction kit can extract soluble plant proteins from fresh, frozen, or dried plant tissues. Suitable for protein extraction from various plants and different parts of plants (such as roots, stems, leaves, flowers, seeds, etc.), with high extraction efficiency, high protein yield, high activity, and fast speed. The extracted protein can be directly subjected to protein electrophoresis analysis, immunoprecipitation, Western Blot, protein activity determination, and protein purification experiments. The concentration of the extracted protein can be determined using the BCA protein quantification kit. P665757Component100 TStorageP665757APlant Protein Extraction Reagent100 mLRTP665757BProtease Inhibitor Cocktail (100×)1 mL-20℃. Avoid freeze/thaw cycle. Precautions：1. This product contains 1mM EDTA.2. To prevent protein degradation, all operations should be carried out on ice as much as possible.3. After extracting protein using this product, the BCA method can be used for protein quantification.4. To achieve the best experimental results, please adjust the optimal usage amount according to the experiment.Operation steps:1. Please remove the required Plant Protein Extraction Agent for pre cooling before protein extraction.2. Weigh the weight of the experimental plant tissue. Add 5 ml of Plant Protein Extraction Agent to 1 g of tissue (add Protein Inhibitor Cocktail in a 1:99 ratio before protein extraction).Attention:1) Before homogenization, cut large pieces of plant tissue into small pieces and homogenize them with a mechanical homogenizer for 10 seconds, with an interval of 10 seconds. Repeat the process three times and select the appropriate homogenization method according to the different tissue samples.2) The amount of lysate used is adjusted according to different parts of the plant. If concentrated protein extracts are needed, the amount of Plant Protein Extraction Agent used can be appropriately reduced.3. After homogenization, incubate on ice for 20-30 minutes.4.4 ℃ 13400 × g, centrifuge for 20 minutes.5. Collect soluble proteins from the supernatant for further purification or downstream analysis... Read More |