| Description | Background information 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 Background information 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). We offer recombinant Annexin V conjugated to a numerous fluorophores, as well as an Annexin V biotin conjugate which can be detected with fluorophore-labeled streptavidin. By binding to PS, fluorophores labeled Annexin V can be used to detect and quantify apoptotic cells via flow cytometry or fluorescence microscopy. The excitation and emission maxima of the Annexin V conjugates are summarized in the following table. Cat.No.Ex/Em (nm)Format1rp226056NABiotin2rp225998343/441AF3503rp225999401/422AF4054rp226057490/525AF4885rp226001590/618AF5946rp226060650/668AF6477rp226002681/704AF6808rp226003752/776AF7509rp226053498/517FITC10rp226004410/455Pacific Blue11rp226006647/665Cy512rp226054650/660APC13rp226055565/575PE14rp226058565/670PE-Cy515rp226059565/774PE-Cy7Precautions1. Please try to avoid light when using to slow down the quenching of fluorescence.2. Propidium Iodide Solution is toxigenic and mutagenic; handle with care.3. Due to the calcium dependence of the Annexin V:PS interaction, it is critical to avoid buffers containing EDTA or other calcium chelators during Annexin V experiments.Instruction for use1. Dilute 10X Binding Buffer (A1372288) 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 amounts of cells in Annexin V Binding Buffer at a concentration of 1.0-5.0 x 106 cells/mL.3. Add 5 µL of Annexin V-AF350 to 100 µL of the cell suspension. Stain with a viability dye, such as PI (P1373641; P1372285), 7-AAD (A1372406), or DAPI (D1372407) dyes, if desired.4. Gently vortex the cells and incubate for 10 min at RT (25°C) in the dark.5. Add 100 µL of 1X Binding Buffer to each assay. Analyze by flow cytometry within 1 hr... Read More | Biochemical Test:SDS-PAGE (purity> 80%); Western blot with patient sample.Calculated Isoelectric Point:pH 8.83 | IRE1α kinase-IN-2 is a potent IRE1α kinase inhibitor, with an EC 50 of 0.82 µM. IRE1α kinase-IN-2 inhibits IRE1α kinase autophosphorylation (IC 50 =3.12 µM). IRE1α kinase-IN-2 inhibits XBP1 mRNA splicing in the WT cell lines.In VitroIRE1α kinase-IN-2 (compoundIRE1α kinase-IN-2 is a potent IRE1α kinase inhibitor, with an EC 50 of 0.82 µM. IRE1α kinase-IN-2 inhibits IRE1α kinase autophosphorylation (IC 50 =3.12 µM). IRE1α kinase-IN-2 inhibits XBP1 mRNA splicing in the WT cell lines.In VitroIRE1α kinase-IN-2 (compound 3) inhibits XBP1 mRNA splicing, even during ER stress. MCE has not independently confirmed the accuracy of these methods. They are for reference only.Form:Solid... Read More | Inquire | Keratinocyte growth factor (KGF) is a cytokine found by Rubin et al. (1989) from the culture supernatant of embryonic lung fibroblasts, which is a member of the FGF family, namely FGF-7. KGF is an effective epithelial-specific growth factor secreted by mesenchymal cells and distributed in epithelialKeratinocyte growth factor (KGF) is a cytokine found by Rubin et al. (1989) from the culture supernatant of embryonic lung fibroblasts, which is a member of the FGF family, namely FGF-7. KGF is an effective epithelial-specific growth factor secreted by mesenchymal cells and distributed in epithelial cells. Its mitotic activity is mainly manifested in keratinocytes, which can specifically promote the proliferation, migration and differentiation of epithelial cells. It is closely related to organ development, wound repair, tumor genesis and immune reconstruction.Activity definition: The ED50 value is less than 1.0 ng/ml, that is, the corresponding activity unit is greater than or equal to 1 x 10*6 units/mg, as determined by the proliferation method of cultured MCF-7 cells... Read More |