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ANNEXIN V:Biotin ASSAY KIT

MBS239211

100 Tests

480 USD

Kits

ANNEXIN V:Biotin ASSAY KIT

ANNEXIN V KIT

annexin A5; Annexin A5; annexin A5; CBP-I; PAP-I; VAC-alpha; anchorin CII; annexin V; annexin-5; calphobindin I; endonexin II; epididymis secretory protein Li 7; lipocortin V; placental anticoagulant protein 4; placental anticoagulant protein I; thromboplastin inhibitor; vascular anticoagulant-alpha; annexin A5; Anchorin CII; Annexin V; Annexin-5; Calphobindin I; CBP-I; Endonexin II; Lipocortin V; Placental anticoagulant protein 4; PP4; Placental anticoagulant protein I; PAP-I; Thromboplastin inhibitor; Vascular anticoagulant-alpha; VAC-alpha

ANXA5; ANXA5; PP4; ANX5; ENX2; RPRGL3; HEL-S-7; ANX5; ENX2; PP4; CBP-I; PP4; PAP-I; VAC-alpha

ANXA5_HUMAN

320

Biotin

Store at 4 degree C. DO NOT FREEZE. This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use. Shelf Life: 12 months from date of despatch.

Flow cytometry (FC/FACS)

Flow Cytometry: Maximum Dilution: Neat

Reagents In The Kits: Annexin V:Biotin 1 x 0.5ml vial, Propidium Iodide 1 x 1.6ml vial at 20ug/ml, Binding buffer 1 x 50ml vial at x 4 concentrate. Note: This assay also requires streptavidin:FITC conjugate for visualisation (not supplied - see recommended useful reagents section). Perservative Stabilisers: 0.02% Sodium Azide (NaN3). 1% Bovine Serum Albumin . Instructions For Use: 1) Dilute the binding buffer 1:4 in distilled water (50ml binding buffer + 150ml distilled water).2) Wash cells in PBS by gentle shaking or pipetting up and down. 3) Resuspend cells in 200ul pre-diluted binding buffer, adjusting to a cell density of 2-5 x 105 cells/ml. 4) Add 5ul Annexin V:Biotin to 195ul of the cell suspension prepared in step 3. 5) Mix and incubate for 15 minutes at room temperature. 6) Wash cells twice in 190ul of pre-diluted binding buffer.7) Resuspend cells in 190ul pre-diluted binding buffer.8) Add streptavidin:FITC conjugate.9) Mix and incubate for 30 minutes in the dark, at room temperature.10) Wash cells in 200ul pre-diluted binding buffer.11) Resuspend cells in 190ul pre-diluted binding buffer. 12) Add 10ul of the Propidium Iodide solution. 13) Analyse by flow cytometry. The flow cytometer is preferably set such that the Mean Fluorescence Intensity of the Annexin V negative population is between 1 and 10. Optimal parameter settings can be found using a positive control. For a positive control, incubate the cells with 3% formaldehyde in buffer during 30 minutes on ice. Wash away the formaldehyde and suspend the cells in cold binding buffer at 2-5 x 105 cells/ml. Proceed with step 2 as described above.

This test employs the property of Annexin V:Biotin to bind to the membrane phospholipid phosphatidylserine (PS) in the presence of Ca2+. PS is exposed at the cell surface during the early stages of apoptosis. Detection of PS is a very sensitive method for detecting cells entering apoptosis, at a time point considerably ahead of nuclear changes such as DNA degradation. The conjugation protocol used to prepare Annexin V:Biotin has not changed the native phospholipid binding properties of Annexin V. This protocol is designed to measure apoptosis easily and quickly in a sample of suspended cells.

4502107

NP_001145.1

NM_001154.3

P08758

35,937 Da

Prostaglandin Synthesis And Regulation Pathway (198912)

The protein encoded by this gene belongs to the annexin family of calcium-dependent phospholipid binding proteins some of which have been implicated in membrane-related events along exocytotic and endocytotic pathways. Annexin 5 is a phospholipase A2 and protein kinase C inhibitory protein with calcium channel activity and a potential role in cellular signal transduction, inflammation, growth and differentiation. Annexin 5 has also been described as placental anticoagulant protein I, vascular anticoagulant-alpha, endonexin II, lipocortin V, placental protein 4 and anchorin CII. The gene spans 29 kb containing 13 exons, and encodes a single transcript of approximately 1.6 kb and a protein product with a molecular weight of about 35 kDa. [provided by RefSeq, Jul 2008]

ANXA5: a calcium/phospholipid-binding protein and an anticoagulant protein that acts as an indirect inhibitor of the thromboplastin-specific complex, which is involved in the blood coagulation cascade. The binding of labeled ANXA5 to phosphatidylserine is used as a marker of apoptosis. Annexins are a family of structurally related proteins whose common property is calcium-dependent binding to phospholipids. There are at least ten different annexins in mammalian species. Annexins do not contain signal peptides, yet some annexins (A1, A2 and A5) appear to be secreted in a physiologically regulated fashion. Protein type: Inhibitor; Apoptosis; Lipid-binding; Calcium-binding. Chromosomal Location of Human Ortholog: 4q27. Cellular Component: focal adhesion; membrane; cytoplasm; intracellular; external side of plasma membrane. Molecular Function: phospholipase inhibitor activity; calcium-dependent phospholipid binding; phospholipid binding; calcium ion binding. Biological Process: response to organic substance; negative regulation of catalytic activity; signal transduction; blood coagulation; negative regulation of coagulation; negative regulation of apoptosis. Disease: Pregnancy Loss, Recurrent, Susceptibility To, 3

100 Tests

480 USD