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|Antigen||Vimentin (VIM) Antibodies|
|Reactivity||Dog (Canine), Human, Mouse (Murine), Rat (Rattus) Alternatives|
|Conjugate||This Vimentin antibody is un-conjugated Alternatives|
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
|19 references available|
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Product Details anti-Vimentin AntibodyTarget Details Vimentin Application Details Handling References for anti-Vimentin antibody (ABIN152563) Images
|Purification||Ammonium sulfate precipitation|
|Immunogen||Recombinant human Vimentin purified from E. coli. [Swiss-Prot# P08670]|
Target Details VimentinProduct Details anti-Vimentin Antibody Application Details Handling References for anti-Vimentin antibody (ABIN152563) Images back to top
|Alternative Name||Vimentin (VIM Antibody Abstract)|
|Background||Gene Symbol: VIM|
|Molecular Weight||Theoretical MW: 50 kDa|
|Pathways||Caspase Cascade in Apoptosis|
Application DetailsProduct Details anti-Vimentin Antibody Target Details Vimentin Handling References for anti-Vimentin antibody (ABIN152563) Images back to top
|Application Notes||Western Blot 1:10000-1:20000, Immunohistochemistry 1:150, Immunocytochemistry/Immunofluorescence 1:5000, Immunohistochemistry-Paraffin 1:150, Immunohistochemistry-FrozenThis Vimentin antibody is useful for Immunohistochemistry paraffin embedded sections, Western blot and Immunocytochemistry/Immunofluorescence. Use in Immunohistochemistry-Frozen reported in scientific literature (PMID 25626686) The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.|
The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.
Immunostaining of cells in tissue culture protocol specific for Vimentin Antibody Immunostaining of cells in tissue cultureThe purpose of fixation is to denature the components of cells enough so that they stay on the dish and can be bound by antibodies, hopefully without destroying cellular morphology. Fixatives such as formalin, paraformaldehyde and glutaraldehyde chemically cross-link proteins, by binding to amino acid side chains. This chemical modification can also have the consequence of blocking antibody binding sites. Substances such as acetone and methonal are not true fixatives but are denaturants, which precipitate proteins without covalently modifying them. We routinely use a combination of mild formalin fixation followed by cold methanol for neurons, mixed neuron/glial cultures and most used cell lines. The formalin preserves the cellular morphology quite well, while the methanol further denatures the proteins of the cells and helps keep what is left of the cell adherent to the dish. For soluble proteins it may be necessary to skip the methanol step, but in this case you have to be very careful with the washing steps, as the cells tend to wash off the dish. Certain antibodies may be very sensitive to formalin fixation, so you may have to experiment a little, perhaps skipping that step. The following procedure work for antibodies to most cytoskeletal and signaling molecules. This procedure is good for cells in 6 well dishes or in 35mm tissue culture plates.
1. Draw the culture medium with aspirator and add 1 mL of 3.7 % formalin in PBS solution to the dish (from 10mls Fisher 37 % formalin plus 90mls PBS, the Fisher formalin contains 37 % formaldehyde plus about 1 % methanol which may be relevant sometimes). Let sit at room temp for 1 minute (can add 0.1 % Tween 20 to PBS used here and all subsequent steps to reduce background, probably best not to do this first time around, though, as it may extract your antigen or help wash your cells off the dish).
. Take off the formalin/PBS and add 1ml of cold methanol (-20C, kept in well sealed bottle in fridge). Let sit for no more than 1 minute.
. Take off methanol and add 1ml of PBS, not letting the specimen dry out. To block nonspecific antibody binding can add ~10ml (=1 %) of goat serum (Sigma), and can incubate for 30 minutes. Then add antibody reagents. Typically 100ml of hybridoma tissue culture supernatent or 1ml of mouse ascites fluid or crude serum. Incubate for 1 hour at room temp. (or 37C for 30 minutes to 1 hour, or 4C overnight, exact time not too critical). Shake gently for well adherent cell lines (3T3, HEK293, etc.).
. Remove primary antibody and replace with 1 mL of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS.
. Add 0.5 mLs of secondary antibody. These are fluorescently labeled goat anti-chicken antibodies and are conjugated to ALEXA dyes and are from Molecular probes (Eugene Oregon, the ALEXA dyes are sulphonated rhodamine compounds and are much more stable to UV than FITC, TRITC, Texas red etc.). Typically make 1:2,000 dilutions of these secondaries in PBS plus 1 % goat serum, BSA or non fat milk carrier. Incubate for 1 hour at room temp. (or 37C for 30 minutes to 1 hour, or 4C overnight). Shake gently for well adherent cell lines (3T3, HEK293, etc.).
. Remove secondary antibody and replace with 1 mL of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS.
. Drop one drop of Fisher mounting medium onto dish and apply 22mm square coverslip. View in the microscope.
|Restrictions||For Research Use only|
HandlingProduct Details anti-Vimentin Antibody Target Details Vimentin Application Details References for anti-Vimentin antibody (ABIN152563) Images back to top
Buffer contains: 5 mM 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||Avoid freeze-thaw cycles|
|Storage||4 °C,-20 °C|
|Storage Comment||Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.|
References for anti-Vimentin antibody (ABIN152563)Product Details anti-Vimentin Antibody Target Details Vimentin Application Details Handling Images back to top
|Product cited in:||
Smith, Hammers, Sweeney, Barton: "Increased collagen cross-linking is a signature of dystrophin-deficient muscle." in: Muscle & nerve, Vol. 54, Issue 1, pp. 71-8, 2016 Further details: Immunohistochemistry (Frozen Sections)
Nakamura, Koshiba-Takeuchi, Tsuchiya, Kojima, Miyazawa, Ito, Ogawa, Takeuchi: "Expression analysis of Baf60c during heart regeneration in axolotls and neonatal mice." in: Development, growth & differentiation, Vol. 58, Issue 4, pp. 367-82, 2016 (Sample species: Mouse (Murine)). Further details: Immunocytochemistry,Immunofluorescence
Behrens, Solga, Ziemann, Rastetter, Berwanger, Herrmann, Noegel, Clemen: "Coronin 1C-free primary mouse fibroblasts exhibit robust rearrangements in the orientation of actin filaments, microtubules and intermediate filaments." in: European journal of cell biology, Vol. 95, Issue 8, pp. 239-51, 2016 (Sample species: Mouse (Murine)). Further details: Immunocytochemistry,Immunofluorescence
Marcuzzo, Bonanno, Kapetis, Barzago, Cavalcante, DAlessandro, Mantegazza, Bernasconi: "Up-regulation of neural and cell cycle-related microRNAs in brain of amyotrophic lateral sclerosis mice at late disease stage." in: Molecular brain, Vol. 8, pp. 5, 2015 (Sample species: Mouse (Murine)). Further details: Immunohistochemistry (Frozen Sections)
Morissette Martin, Maux, Laterreur, Mayrand, L Gagné, Moulin, Fradette: "Enhancing repair of full-thickness excisional wounds in a murine model: Impact of tissue-engineered biological dressings featuring human differentiated adipocytes." in: Acta biomaterialia, Vol. 22, pp. 39-49, 2015 (Sample species: Mouse (Murine)). Further details: Immunohistochemistry (Frozen Sections)
Wong, Chen, Huang, Sehba, Friedel, Zou: "Attenuation of Cerebral Ischemic Injury in Smad1 Deficient Mice." in: PLoS ONE, Vol. 10, Issue 8, pp. e0136967, 2015 (Sample species: Mouse (Murine)). Further details: Immunohistochemistry (Frozen Sections)
Ge, Pettan-Brewer, Morton, Carter, Fatemi, Rabinovitch, Ladiges: "Mitochondrial catalase suppresses naturally occurring lung cancer in old mice." in: Pathobiology of aging & age related diseases, Vol. 5, pp. 28776, 2015 (Sample species: Mouse (Murine)). Further details: Immunohistochemistry
Pócsai, Kálmán: "Glial and perivascular structures in the subfornical organ: distinguishing the shell and core." in: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 63, Issue 5, pp. 367-83, 2015 Method employed by authors: Western Blotting (WB)
Busch, Moser, Gurley, Kelly-Spratt, Liggitt, Kemp: "ARF inhibits the growth and malignant progression of non-small-cell lung carcinoma." in: Oncogene, Vol. 33, Issue 20, pp. 2665-73, 2014 (Sample species: Mouse (Murine)). Further details: Immunohistochemistry (Paraffin-embedded Sections)
Phamduong, Rathore, Crews, DAngelo, Leinweber, Kappera, Krenning, Rendell, Belcheva, Coscia: "Acute and chronic mu opioids differentially regulate thrombospondins 1 and 2 isoforms in astrocytes." in: ACS chemical neuroscience, Vol. 5, Issue 2, pp. 106-14, 2014 (Sample species: Rat (Rattus)). Further details: Immunocytochemistry,Immunofluorescence
Muir, Lim, Benitez, Modayur Chandramouleeswaran, Lee, Ruchelli, Spergel, Wang: "Esophageal epithelial and mesenchymal cross-talk leads to features of epithelial to mesenchymal transition in vitro." in: Experimental cell research, Vol. 319, Issue 6, pp. 850-9, 2013 (Sample species: Human). Further details: Immunocytochemistry,Immunofluorescence,Immunohistochemistry
Stadler, Vincent, Fedorov, Patsialou, Cherrington, Wakshlag, Mohanan, Zee, Zhang, Garcia, Condeelis, Brown, Coonrod, Allis: "Dysregulation of PAD4-mediated citrullination of nuclear GSK3β activates TGF-β signaling and induces epithelial-to-mesenchymal transition in breast cancer cells." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, Issue 29, pp. 11851-6, 2013 (Sample species: Human). Further details: Immunocytochemistry,Western Blotting,Immunofluorescence
Murray, Mendez, Janmey: "Substrate stiffness regulates solubility of cellular vimentin." in: Molecular biology of the cell, Vol. 25, Issue 1, pp. 87-94, 2013 (Sample species: Human). Further details: Immunocytochemistry,Western Blotting,Immunofluorescence
Zamanian, Xu, Foo, Nouri, Zhou, Giffard, Barres: "Genomic analysis of reactive astrogliosis." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 32, Issue 18, pp. 6391-410, 2012 (Sample species: Mouse (Murine)). Further details: Immunocytochemistry,Immunofluorescence,Immunohistochemistry
Ponzo, Lesurf, Petkiewicz, OMalley, Pinnaduwage, Andrulis, Bull, Chughtai, Zuo, Souleimanova, Germain, Omeroglu, Cardiff, Hallett, Park: "Met induces mammary tumors with diverse histologies and is associated with poor outcome and human basal breast cancer." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 106, Issue 31, pp. 12903-8, 2009
Johnstone, Mongroo, Rich, Schupp, Bowser, Delemos, Tobias, Liu, Hannigan, Rustgi: "Parvin-beta inhibits breast cancer tumorigenicity and promotes CDK9-mediated peroxisome proliferator-activated receptor gamma 1 phosphorylation." in: Molecular and cellular biology, Vol. 28, Issue 2, pp. 687-704, 2008 (Sample species: Human). Further details: Immunohistochemistry
Okawa, Michaylira, Kalabis, Stairs, Nakagawa, Andl, Johnstone, Klein-Szanto, El-Deiry, Cukierman, Herlyn, Rustgi et al.: "The functional interplay between EGFR overexpression, hTERT activation, and p53 mutation in esophageal epithelial cells with activation of stromal fibroblasts induces tumor development, invasion, and ..." in: Genes & development, Vol. 21, Issue 21, pp. 2788-803, 2007 (Sample species: Human). Further details: Western Blotting
Rankin, Tomaszewski, Haase: "Renal cyst development in mice with conditional inactivation of the von Hippel-Lindau tumor suppressor." in: Cancer research, Vol. 66, Issue 5, pp. 2576-83, 2006 (Sample species: Mouse (Murine)). Further details: Immunohistochemistry
Anstine, Horne, Horwitz, Lincoln: "Contribution of Extra-Cardiac Cells in Murine Heart Valves is Age-Dependent." in: Journal of the American Heart Association, Vol. 6, Issue 10, 1970 Method employed by authors: Immunofluorescence (fixed cells) (IF/ICC) (Sample species: Mouse (Murine)).
ImagesProduct Details anti-Vimentin Antibody Target Details Vimentin Application Details Handling References for anti-Vimentin antibody (ABIN152563) back to top