Annexin V Antibody Review

Introduction

a marker of cells in an early stage of apoptosis. PMID: 11708469, PMID: 9298227
an in vivo marker of cellular injury and death. PMID: 14985868
a calcium-dependent phospholipid-binding protein. PMID: 15173196
an intracellular platelets glycoprotein. PMID: 10174688
has an important role in the regulation of apoptosis. PMID: 14616799
play a role in developmental processes such as cardiac growth processes. PMID: 9546604
binds phosphatidylserine that becomes exposed on the cell membrane in apoptotic cells. PMID: 16813956, PMID: 12920851
has a high affinity for exposed phosphatidylserine on apoptotic cells. PMID: 14676140
pivotal in the maintenance of pregnancy by preventing the activation of blood coagulation. PMID: 15175799
can be used to detect apoptotic cells in vitro and in vivo, based on its ability to identify extracellular phosphatidylserine, which arises during apoptosis. PMID: 14666384

Normal Expression

Annexin V content in the lungs and myocardium in normal rats was extremely high in comparison to that in brain and skeletal muscle. PMID: 12745185
A potent anticoagulant, annexin V, is abundant in placental tissues. PMID: 10203671
Immunohistochemical analysis of Anx V in growth plate cartilage, confirmed by in situ hybridization, suggests that Anx V is prominently expressed and forms a major constituent of growth plate chondrocytes. Anx V epitopes are also located in the pericellular matrix of hypertrophic cartilage. In adult articular cartilage the expression is downregulated, with the highest levels of immunostaining found in the upper third of the articular cartilage layers and almost no antigen found in the deep layers. Osteoarthritic (OA) cartilage is characterized by a significant upregulation of message and protein throughout the entire depth of the tissue, an accumulation of cytoplasmic annexin V epitopes, and a release of epitopes into the pericellular and interterritorial matrix, in part co-localized with granular structures. PMID: 9889256
Annexin V staining was observed in all arrested and/or fragmented human embryos, but not in cryopreserved embryos which continued to develop normally after thawing. PMID: 9733435
Annexin V is associated with the sarcolemma and intercalated discs of cardiac myocytes in sections of adult porcine and rat heart. Annexin V associates tightly with the sarcolemma and suggests that components in addition to phospholipid are involved in binding annexin V to the membrane. In these differentiating neonatal cells, annexin V is also located in the nucleoplasm and at the periphery of the nucleus. PMID: 9299563
Immunocytochemistry revealed that annexin V was localized in the nucleus and throughout the cytoplasm in human foreskin fibroblasts. PMID: 8843166
Since annexin V has been reported as an inhibitor of protein kinase C, its nuclear localization in association with the internal matrix, which plays an important role in several nuclear processes, indicates its involvement in the regulation of signal transduction. PMID: 8753782
annexin V was stained as a striated pattern along myofibrils on frozen sections of both atria and ventricles of adult rats. The striations were seen more clearly in cultured rat atrial myocytes. Examination of doubly stained cardiocytes with anti-annexin V and anti-alpha-actinin by a confocal laser scanning microscope suggests that annexin V is localized in association with the Z-line of rat cardiac myocytes. PMID: 7585827
Annexin V was found to localize to the microvillar surface of the villous syncytiotrophoblasts. PMID: 7824446
Annexin V is an intracellular protein recently shown to be localized to nucleoli and cytosol. In this study we show that cytosolic annexin V is associated with mitochondria. PMID: 8354413
found diffusely in the cytoplasm and associated with plasma membranes, membranes delimiting cytoplasmic vacuoles, membranes of the endoplasmic reticulum, and filamentous structures the identity of which remains to be established. PMID: 8495746
Immunofluorescent localization studies indicate that in primary cultures the protein is abundant in the cytoplasm of the cells and also extends into the nucleus. PMID: 8484739
Indirect immunofluorescence and immunoblotting studies using affinity-purified anti-annexin V antibodies revealed that annexin V is located within the cytoplasm and nucleus of these cells. PMID: 1468579
in the cerebellum the protein is restricted to glial cells, where it is found diffusely in the cytoplasm as well as associated with plasma membranes. Bergmann glial cell bodies and processes and astrocytes in the cerebellar cortex and oligodendrocytes in the cerebellar white matter displayed an intense immune reaction product. In sciatic nerves, the protein was exclusively found in Schwann cells with a subcellular localization similar to that seen in glial cells in the cerebellum. Pituicytes in the neurohypophysis were intensely immunostained, whereas axons were not. In the heart, annexin V was restricted to the sarcolemma, transverse tubules, and intercalated discs. In skeletal muscles the protein was localized to the sarcolemma and transverse tubules. PMID: 1387136
Glial cells in the cerebellum and in the optic nerve, the corneal epithelium, the posterior epithelium in the iris, chondrocytes, skeletal muscle cells and cardiomyocytes, the capillary endothelial cells in many organs, the muscularis mucosae and the muscular layer in the intestinal tract, hepatocytes, Muller cells in the retina, the lens fibers, Sertoli and Leydig cells in the testis, and smooth muscle cells in the epididymis and bronchi displayed intense immunostaining. In the adrenal gland, only the cortex showed immunoreaction product. In the kidney, no apparent staining of renal cells was observed, whereas endothelial cells of peritubular capillaries were stained. In the heart, annexin V was found associated exclusively with the sarcolemma and intercalated discs, as opposed to the diffuse distribution of the protein in skeletal muscle cells. In the spleen, only reticular elements in the white pulp and endothelial cells in the red pulp appeared to be immunostained. PMID: 1833446

Abnormal Expression

Annexin V localized to the cytoplasm of injured neurons ipsilateral to the site of injury as well as to otherwise normal-appearing neurons of the contralateral hemisphere as confirmed by dual fluorescent microscopy. PMID: 14985868
a massive number of circulating annexin-V-binding neutrophils in the absence of apoptosis can be demonstrated in Barth syndrome (BTHS). PMID: 14764526
high levels of plasma annexin V in patients with acute myocardial infarction (AMI), cardiac arrest and severe trauma reflect the severity of damage of the myocardium and/or other visceral organs, PMID: 12745185
annexin V was detected on trophoblasts in the placentae and the staining intensity of annexin V in the placentae from such patients was reduced when compared to normal placenta. PMID: 10629367
At electron microscopy, gold-labeling of Annexin V was located on the plasma membrane only of apoptotic thymocytes and on cytoplasmic debris, likely resulting from the typical apoptotic blebbing. PMID: 9359032

Expression Alteration

presence of annexin V in nuclei reflects translocation rather than catabolism and resynthesis. Inhibition of tyrosine kinase activities with genistein attenuates the relocation of annexin V from the cytoplasm to the nucleus. Thus, the nuclear location of annexin V is controlled by signaling pathways involving serum factors and tyrosine kinases. The results argue for an important role for annexin V in the cell nucleus. PMID: 9223374

Function

membrane-binding activity of annexin V is much more sensitive to amine-directed chemical modification than previously realized. New annexin V molecules labeled by site-specific methods will greatly improve sensitivity for detecting cell death in vivo. PMID: 16954565
annexin V has a function in M cell-mediated transcytosis. PMID: 16552796
Overexpression of annexin V is sufficient to stimulate these terminal differentiation events in growth plate chondrocytes, whereas suppression of annexin V expression inhibits these events. PMID: 15777796
[18F] labeled annexin V can be readily prepared by the conjugation of annexin V with [18F]SFB and that the positron-emitting compound is biologically active in detecting apoptosis. PMID: 15246365
a potential role for 99mTc-Annexin-V (ANX) imaging in the diagnosis of myocarditis. PMID: 15181144
Anx V helps to prevent clotting in the placenta during pregnancy. PMID: 15175799
regulates coagulability in the blood stream by binding not only to phosphatidylserine but also to sulfatide. PMID: 15173196
inhibits prothrombin activation and is able to prevent thrombus formation under normal venous and arterial blood flow conditions. PMID: 15652780
a human protein with a high affinity for phosphatidylserine, labeled with (99m)Tc can detect apoptosis in vivo. PMID: 14960654
a calcium-dependent phospholipid-binding protein that exhibits anticoagulant activity on binding to phosphatidylserine exposed on the activated surfaces of endothelial cells and platelets, inhibiting activation of factor X and prothrombin in the blood coagulation cascade. PMID: 15173196
an in vivo marker of cellular injury and death, could be used to noninvasively monitor neuronal injury following focal middle cerebral artery (MCA) occlusion/reperfusion injury. PMID: 14985868
Because annexin V clears rapidly from blood and targets apoptotic macrophage population, it should constitute an attractive imaging agent for the noninvasive detection of unstable atherosclerotic plaques. PMID: 14676140
in newt testis annexin V is a cold-sensitive protein, suggesting the possibility that annexin V might have a cold stress-related function in newt germ cells. PMID: 12832824
the Annexin-V binding assay was able to detect deleterious changes in the sperm plasma membrane at an earlier point than PI staining, thus representing a novel approach to investigating membrane integrity in this species. PMID: 12832017
labeled annexin-V is useful for in situ detection of cell death in an in vivo model of I/R in the heart and for the evaluation of cell death-blocking strategies. PMID: 11004148
AV binding is an excellent marker for apoptotic cells, but that these cells already have fragmented DNA. PMID: 10969816
Anx-V-biotin labels dental cells in early stages of cell death and indirectly cells that have ingested labelled apoptotic cells during the course of the experiment. PMID: 10857481
annexin-V binding assay represents a powerful new flow cytometric method to monitor mast cell degranulation for functional analysis. PMID: 10404150
When the cell membrane is disturbed, phospholipid phosphatidylserine (PS) is translocated from the inner to the outer leaflet of the plasma membrane. This is one of the earliest signs of apoptosis and can be monitored by the calcium-dependent binding of annexin V. PMID: 10065865
Annexin V binding assay as a tool to measure apoptosis in differentiated neuronal cells. PMID: 9894786

Applications

Electron Microscopy (EM)

Application of biotinilated annexin V for EM detection with gold-conjugated anti-biotin, showed that only clear-cut apoptotic, apoptotic necrotic and oncotic cells showed the gold-label at their membranes. PMID: 11773704
we describe a method for the immunogold-labeling of biotin-conjugated Annexin V, to detect apoptotic thymocytes at electron microscopy. PMID: 9359032
Both immunoblot and immunoelectron microscopic analyses with intact tissues, synaptosomes and purified synaptic vesicles showed that annexin V was expressed in neurons, preferentially concentrated in axon terminals and associated with synaptic vesicles. PMID: 8931014
We evaluated the histological and ultrastructural localization of the potent anticoagulant protein, annexin V, at the light and electron microscopic levels, using immunohistochemistry and an immunogold method. PMID: 7824446
ultrastructural localization of annexin V a Ca(2+)-dependent phospholipid- and membrane-binding protein in the nervous system, heart, and skeletal muscles. PMID: 1387136

ELISA

A serum sample was taken from 1038 consecutive healthy women at the beginning of pregnancy. IgG and IgM anti-AnxV antibodies were measured by an ELISA method. PMID: 16014551
To evaluate the role of anti-annexin V antibodies (AA5A) in Takayasu's arteritis (TA), we investigated these antibodies in the sera of 66 TA patients, 50 healthy controls and in the follow-up sera of 12 active TA patients by enzyme-linked immunosorbent assay. PMID: 14616799
localization of annexin V, a calcium binding protein, was immunochemically and immunohistologically studied in experimental rat glomerulonephritis using annexin V polyclonal antibody. Plasma and urinary annexin V levels were measured by a sandwich enzyme-linked immunosorbent assay (ELISA). PMID: 11271515
Measurement of urinary annexin V by ELISA and its significance as a new urinary-marker of kidney disease. PMID: 10876002
Measurement of plasma annexin V by ELISA in the early detection of acute myocardial infarction. PMID: 8814351

Flow Cytometry (FC)

Flow cytometric analysis of agonist-induced annexin V, factor Va and factor Xa binding to human platelets. PMID: 16793697
propidium-iodide and annexin-V index using flow-cytometry is a quick, useful, and easily accessible method for the evaluation of plasma cell kinetics and thus prognosis of the disease, multiple myeloma. PMID: 16601769
Flow cytometry was used to assess endothelial cell apoptosis by quantification of circulating CD31+/annexin V+ apoptotic microparticles in peripheral blood. PMID: 16239600
Annexin V binding to human umbilical vein endothelial cells (HUVECs) as determined by flow cytometry after 24-hour culture with plasma was decreased when plasma from SLE cases was used (SLE cases versus population controls: P=0.002; SLE cases versus SLE controls P=0.02). PMID: 15539620
annexin V binding by leukocytes was determined daily using flow cytometry and FITC-labeled annexin V in 33 postoperative patients with severe sepsis or septic shock during their intensive care unit stay. PMID: 15033821
Plasma cell proliferative activity was measured by means of a propidium iodide index (PC-PI) examined by flow cytometry using a DNA/CD138 double staining technique. For detection of plasma cells entering apoptosis (PC-AI) flow cytometry method with annexin V FITC and MoAb CD138 was used. PMID: 14628090
The cells of Burkitt lymphoma (BL) cell line Raji were incubated with 1.0 micromol/L dexamethasone (DEX) for 2, 4 and 8 h respectively, then stained with Annexin V-FITC (fluorescein isothiocyanate conjugated) which was used to detect the exposed phosphatidylserine (PS) on the epimembrane resulting from a loss of phospholipid asymmetry in the early stage of apoptosis, and also stained with propidium iodide (PI) which allows analysis of secondary necrotic cells related with cell membrane and DNA damage that probably representlate stage of apoptosis, then apoptotic cells were quantified by flow cytometry (FCM). Annexin-V assay is a specific, sensitive, accurate, reproductive and quantitative method for analyzing apoptotic cells. PMID: 12894879
present study evaluated the ability of an Annexin-V binding assay to detect early changes in sperm membrane integrity using flow cytometry (FC) in two different portions of the boar ejaculate, in cryopreserved semen. PMID: 12832017
Comparison of the TUNEL, lamin B and annexin V methods for the detection of apoptosis by flow cytometry. PMID: 12553704
Analysis of radiation-induced apoptosis in human lymphocytes: flow cytometry using Annexin V and propidium iodide versus the neutral comet assay. PMID: 12116376
The dynamic process of apoptosis analyzed by flow cytometry using Annexin-V/propidium iodide and a modified in situ end labeling technique. PMID: 11774346
PS exposure was measured flow cytometrically using FITC-labelled annexin V, combined with PI. PMID: 11773704
Apoptotic index by Annexin V flow cytometry: adjunct to morphologic and cytogenetic diagnosis of myelodysplastic syndromes. PMID: 11241504
Flow cytometric analysis of apoptotic subpopulations with a combination of annexin V-FITC, propidium iodide, and SYTO 17. PMID: 11169578
Flow cytometric estimation of the plasma membrane diversity of transplantable melanomas, using annexin V. PMID: 10763124
Measurement of phosphatidylserine exposure in leukocytes and platelets by whole-blood flow cytometry with annexin V. PMID: 10744422
Quantitative measurement of mast cell degranulation using a novel flow cytometric annexin-V binding assay. PMID: 10404150
A novel method for measuring CTL and NK cell-mediated cytotoxicity using annexin V and two-color flow cytometry. PMID: 10357200
annexin V-binding, in conjunction with flow cytometry, was used to evaluate the integrity of the sperm plasma membrane after different cryostorage protocols: i.e. 10% (v/v) glycerol; sperm maintenance medium (MM); freezing medium TEST yolk buffer (TYB); or cryostorage without protection (cryoshock). PMID: 10065865
Use of intracellular pH and annexin-V flow cytometric assays to monitor apoptosis and its suppression by bcl-2 over-expression in hybridoma cell culture. PMID: 9894897
Flow cytometry using annexin V can detect early apoptosis in peripheral blood stem cell harvests from patients with leukaemia and lymphoma. PMID: 9535035
A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. PMID: 7622868
Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. PMID: 8068938

Immunocytochemistry (ICC)

the apoptotic cell population was identified immunocytochemically using Annexin V, a marker of cells in an early stage of apoptosis. PMID: 11708469
Immunocytochemistry revealed that annexin V was localized in the nucleus and throughout the cytoplasm in human foreskin fibroblasts. PMID: 8843166
subcellular distribution of annexin V, a calcium-dependent phospholipid- and membrane-binding protein, in a human-derived cell line, GL15, was investigated by immunocytochemistry at light and electron microscope levels. PMID: 8495746

Immunofluorescence (IF)

In vivo detection of apoptotic cells with fluorescently labeled annexin V is an emerging technique that we evaluated for detecting apoptotic germ cells in a mouse model of testicular torsion. PMID: 16813956
Assaying annexin V binding by indirect immunofluorescence was demonstrated to be more sensitive and faster than the JAM test, which is a well-described, sensitive and simple assay for DNA fragmentation and cell death. PMID: 9776575
Immunofluorescent localization studies indicate that in primary cultures the protein is abundant in the cytoplasm of the cells and also extends into the nucleus. PMID: 8484739
Indirect immunofluorescence and immunoblotting studies using affinity-purified anti-annexin V antibodies revealed that annexin V is located within the cytoplasm and nucleus of these cells. PMID: 1468579

Immunohistochemistry (IHC)

localization of annexin V, a calcium binding protein, was immunochemically and immunohistologically studied in experimental rat glomerulonephritis using annexin V polyclonal antibody. Plasma and urinary annexin V levels were measured by a sandwich enzyme-linked immunosorbent assay (ELISA). PMID: 11271515
Immunohistochemical study of annexin V expression in placentae of preeclampsia. PMID: 10629367
cellular distribution of annexin V (CaBP33) in rat tissues by immunohistochemistry. PMID: 1833446

Western Blot (WB)

sera in patients with scleroderma were examined for IgG anti-annexin V antibodies by ELISA and immunoblotting. PMID: 10529134
Western blot analysis of annexin V expression showed no differences between study patients and control subjects. PMID: 10411817
localization of annexin V and annexin VI during the development of rat fetal limb buds by immunoblot and immunocytochemical analysis.
Both immunoblot and immunoelectron microscopic analyses with intact tissues, synaptosomes and purified synaptic vesicles showed that annexin V was expressed in neurons, preferentially concentrated in axon terminals and associated with synaptic vesicles. PMID: 8931014