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Annexin A5 (ANXA5) Protein

Details for Product No. ABIN412007, Supplier: Log in to see
Protein Name
  • anx
  • Anx5
  • ANX5
  • anx5
  • anxa5
  • ANX V
  • cb989
  • ENX2
  • enx2
  • LC5
  • MGC89158
  • PP4
  • R74653
  • RPRGL3
  • wu:fa98f06
  • wu:fj10f10
Reactivity
44
15
10
3
3
3
3
3
1
1
Host
61
9
8
2
1
1
Application
Fluorescence Microscopy (FM), Flow Cytometry (FACS)
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Purpose Reagent for detecting early stages of apoptosis.
Sample Type Cell Culture
Characteristics The Annexin V-FITC conjugate can be used for detection of apoptosis by fluorescence microscopy or by flow cytometry.
Background During apoptosis, phosphatidylserine (PS) is translocated from the cytoplasmic face of the plasma membrane to the cell surface. Annexin V has a strong, Ca2+-dependent affinity for PS and therefore is used as a probe for detecting apoptosis.
Research Area Cardiovascular, Atherosclerosis, Apoptosis/Necrosis, Coagulation
Pathways Apoptosis
Application Notes Detection method: Flow cytometry (Ex/Em = 488/530 nm) using FL1 channel and fluorescence microscopy
Protocol A. Incubation of cells with Annexin V-FITC:
1. Induce apoptosis by desired methods.
2. Collect 1 x 10^5 cells by centrifugation.
3. Resuspend cells in 500 µL of 1X Annexin V Binding Buffer (ABIN412353).
4. Add 1 µL of Annexin V-FITC and 1 µL Propidium Iodide (ABIN412371).
5. Incubate at room temperature for 5 min in the dark.
Proceed to B or C below depending on method of analysis.

B. Quantification by Flow Cytometry:
Analyze cells by flow cytometry (Ex = 488 nm, Em = 530 nm) using FL1 channel for detecting Annexin v-FITC staining and FL2 channel for detetcing PI staining. For adherent cells, trypsinize and gently wash cells with serum-containing medium before incubation with Annexin V-FITC (A.3-5).

C. Detection by Fluorescence Microscopy:
1. Place the cell suspension from Step A.5 on a glass slide, and cover with a glass coverslip. For analyzing adherent cells, grow cells directly on a coverslip. Following incubation (A.5), invert coverslip on a glass slide and visualize cells. The cells can also be washed with 1X Annexin V Binding Buffer and fixed in 2 % formaldehyebefore visualization. (Cells must be incubated with Annexin V-FITC before fixation because any cell membrane disruption can cause nonspecific binding of annexin V to PS on the inner surface of the cell membrane.)
2. Observe the cells under a fluorescence microscope using a dual filter set for FITC and rhodamine or separate filters. Cells that have bound Annexin V-FITC will show bright green staining on the plasma membrane. Cells which have lost membrane integrity will show red staining (PI) throughout the nucleus and a halo of green staining (FITC) on the cell surface (plasma membrane).


TESTING RESULTS: Jurkat cells (treated with 2 µM camptothecin for 6 hours) were collected for annexin V assay according to the kit instructions. Results show 40-60 % apoptotic cells as analyzed by flow cytometry.
Restrictions For Research Use only
Format Liquid
Concentration 0.15 mg/mL
Buffer PBS containing 1 % BSA, 0.02 % sodium azide
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Do not freeze!
Storage 4 °C
Expiry Date 12 months
Product cited in: Cleophas, Crişan, Lemmers, Toenhake-Dijkstra, Fossati, Jansen, Dinarello, Netea, Joosten: "Suppression of monosodium urate crystal-induced cytokine production by butyrate is mediated by the inhibition of class I histone deacetylases." in: Annals of the rheumatic diseases, 2015

Crișan, Cleophas, Oosting, Lemmers, Toenhake-Dijkstra, Netea, Jansen, Joosten: "Soluble uric acid primes TLR-induced proinflammatory cytokine production by human primary cells via inhibition of IL-1Ra." in: Annals of the rheumatic diseases, 2015

Shetty, Matalia, Nuijts, Subramani, Dhamodaran, Pandian, Jayadev, Das: "Safety profile of accelerated corneal cross-linking versus conventional cross-linking: a comparative study on ex vivo-cultured limbal epithelial cells." in: The British journal of ophthalmology, Vol. 99, Issue 2, pp. 272-80, 2015

Brentnall, Weir, Rongvaux, Marcus, Boise: "Procaspase-3 regulates fibronectin secretion and influences adhesion, migration and survival independently of catalytic function." in: Journal of cell science, Vol. 127, Issue Pt 10, pp. 2217-26, 2014

Gao, Dong, Lin, Sheng, Zhang, Su, Xu: "Reduction of AUF1-mediated follistatin mRNA decay during glucose starvation protects cells from apoptosis." in: Nucleic acids research, Vol. 42, Issue 16, pp. 10720-30, 2014

Gaudette, Iwakoshi, Boise: "Bcl-xL protein protects from C/EBP homologous protein (CHOP)-dependent apoptosis during plasma cell differentiation." in: The Journal of biological chemistry, Vol. 289, Issue 34, pp. 23629-40, 2014

Herrmann, Cherryholmes, Schroeder, Phallen, Alizadeh, Xin, Wang, Lee, Lahtz, Swiderski, Armstrong, Kowolik, Gallia, Lim, Brown, Badie, Forman, Kortylewski, Jove, Yu: "TLR9 is critical for glioma stem cell maintenance and targeting." in: Cancer research, Vol. 74, Issue 18, pp. 5218-28, 2014

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Pal, Cao, Gaisina, Bhattacharya, Dutta, Wang, Gunosewoyo, Kozikowski, Billadeau, Mukhopadhyay: "Inhibition of GSK-3 induces differentiation and impaired glucose metabolism in renal cancer." in: Molecular cancer therapeutics, Vol. 13, Issue 2, pp. 285-96, 2014

Tanaka-Nakanishi, Yasunaga, Takai, Matsuoka: "HTLV-1 bZIP factor suppresses apoptosis by attenuating the function of FoxO3a and altering its localization." in: Cancer research, Vol. 74, Issue 1, pp. 188-200, 2014

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Kim, Park, Kim, Choi, Lee, Lee, Choe, Kim, Jeoung: "miR-200b and cancer/testis antigen CAGE form a feedback loop to regulate the invasion and tumorigenic and angiogenic responses of a cancer cell line to microtubule-targeting drugs." in: The Journal of biological chemistry, Vol. 288, Issue 51, pp. 36502-18, 2013

Liu, Shoji-Kawata, Sumpter, Wei, Ginet, Zhang, Posner, Tran, Green, Xavier, Shaw, Clarke, Puyal, Levine: "Autosis is a Na+,K+-ATPase-regulated form of cell death triggered by autophagy-inducing peptides, starvation, and hypoxia-ischemia." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, Issue 51, pp. 20364-71, 2013

E, Cao, Bhattacharya, Dutta, Wang, Mukhopadhyay: "Endogenous vascular endothelial growth factor-A (VEGF-A) maintains endothelial cell homeostasis by regulating VEGF receptor-2 transcription." in: The Journal of biological chemistry, Vol. 287, Issue 5, pp. 3029-41, 2012

Dondeti, Wubbenhorst, Lal, Gordan, DAndrea, Attiyeh, Simon, Nathanson: "Integrative genomic analyses of sporadic clear cell renal cell carcinoma define disease subtypes and potential new therapeutic targets." in: Cancer research, Vol. 72, Issue 1, pp. 112-21, 2012

Chen, Huang, Fan-Chiang, Liu, Lee: "Apoptosis of Ascogregarina taiwanensis (Apicomplexa: Lecudinidae), which failed to migrate within its natural host." in: The Journal of experimental biology, Vol. 216, Issue Pt 2, pp. 230-5, 2012

Qi, Cooke, Liu, Rimsza, Roe, Persky, Miller, Mahadevan: "Aurora inhibitor MLN8237 in combination with docetaxel enhances apoptosis and anti-tumor activity in mantle cell lymphoma." in: Biochemical pharmacology, 2011