Anti-Vimentin antibody [EPR3776] - BSA and Azide free

33 product images

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  Image from Prestigiacomo V et al. PLoS One. 2017;12(6):e0179995. Fig 7.; doi: 10.1371/journal.pone.0179995.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Immunostaining of formalin fixed paraffin embedded human microtissues after exposure to MTX, TAA and TGF-β1.
  

  Formalin fixed paraffin embedded slides of HepaRG/THP-1 macrophages/hTERT-HSC microtissues were stained with Hematoxylin & Eosin (H&E) and vimentin after 14 days of treatment with MTX, TAA and TGF-β1. Microtissues were fixed in 4% PFA and embedded in 2% agarose prior to paraffinization. Microtissues showed increase in the vimentin positive cells after MTX, TAA and TGF-β1 exposure. Vimentin stainings show proliferation of stellate cells and THP-1 macrophages in the microtissues, suggesting the onset of inflammation process.

  For full image see PMID 28665955.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Vimentin immunohistochemistry staining of rhesus monkey retina tissue using rabbit anti-Vimentin antibody
  
  

  Immunohistochemical staining of paraffin embedded paraformaldehyde fixed rhesus monkey retina tissue with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 (green) at a working dilution of 1/200. The sample was incubaded with the primary antibody fro 20 hours at 4°C in 2.5% serum. The secondary antibody used is a Goat anti-rabbit AlexaFluor 488 at 1/400. Heat mediated antigen retrieval was perfomed using citrate pH 6. Tissue was blocked with 5% serum for 1 hour 30 minutes at 25°C

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Clone EPR3776 (ab193555) has been successfully conjugated by Abcam. This image was generated using Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab185030 for protocol details.

  Alexa Fluor® 488 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab185030 staining Vimentin in wild-type HAP1 cells (top panel) and Vimentin knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 488 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab185030 at a 1/500 dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  This image is courtesy of a customer review submitted by Thomas Read.

  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Unpurified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 staining vimentin in human Schlemms Canal Endothelium cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with Triton X-100 0.2% and blocked with 10% serum for 30 minutes at 20°C. Samples were incubated with primary antibody (1/200 in DPBS) for 3 hours at 20°C. An undiluted Alexa Fluor^®488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Clone EPR3776 (ab193555) has been successfully conjugated by Abcam. This image was generated using Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (PE). Please refer to PE Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab209446 for protocol details.

  PE Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab209446 staining Vimentin in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with PE Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab209446 at 1/500 dilution (Pseudocolored in green) and Alexa Fluor® 647 Anti-Tubulin antibody [YOL1/34] - Microtubule Marker ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).

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  Image from Muniz-Talavera H and Schmidt JV. PLoS One. 2017;12(12):e0184957. Fig 3.; doi: 10.1371/journal.pone.0184957.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Jhy^lacZ/lacZ mice exhibit delayed radial glial to ependymal cell differentiation.

  Immunohistochemical analysis of P10 lateral ventricle coronal sections from Jhy^+/+ (A, E) and Jhy^lacZ/lacZ (I, M) mice for expression of Vimentin (pink, Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547), Glast (green) and Acα-Tub (orange) in dorsal (A-D, I-L) and ventral (E-H, M-P) brain regions. Lower right panels (D, L, H, P) represent a higher magnification view of the merged image. In Jhy^+/+, medial wall dorsal and ventral cells express the differentiated ependymal markers Vimentin (A, B, E, F) and Acα-Tub (A, D, E, H), but are negative for the radial glial marker Glast (A, C, E, G). In Jhy^lacZ/lacZ brains, some dorsal cells remain positive for the undifferentiated marker Glast (I, K), while also expressing the differentiated markers Vimentin and Acα-Tub (I, J, L). Jhy^lacZ/lacZ ventral cells express only Vimentin and Acα-Tub (M-P). The dotted line indicates the medial wall ependymal cells in (C, G, K, O). (Q-R) Graphical representation of the percentage of Glast(-)Vimentin(+)Acα-Tub(+) (black bar) and Glast(+)Vimentin(+)Acα-Tub(+) (grey bar) cells in dorsal (Q) and ventral (R) ependymal cells. MW, medial wall; LW, lateral wall; LV, lateral ventricle; * denotes p≤0.05. Scale bars: 50μm (A-P).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Clone EPR3776 (ab193555) has been successfully conjugated by Abcam. This image was generated using Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab194719 for protocol details.

  Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab194719 staining Vimentin in NIH3T3 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab194719 at 1/100 dilution(shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Western blot - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  This data was developed using Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547, the same antibody clone in a different buffer formulation. Different batches of Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 were tested on HEK-293 (Human embryonic kidney epithelial cell) lysate at 0.02 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 54 kDa.

  All lanes: Western blot - Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547)

  Predicted band size: 53 kDa

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  Flow Cytometry (Intracellular) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Overlay histogram showing HAP1 wildtype (green line) and HAP1-VIM knockout cells (red line) stained with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547, 0.5μg/ml) for 30 min at 22°C. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody at 1/2000 dilution for 30 min at 22°C. A Rabbit IgG isotype control antibody (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-VIM knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).
  

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  This image is courtesy of an anonymous customer review.

  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 staining Vimentin in human adenocarcinoma cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% TX-100 in PBS and blocked with 5% serum for 1 at 21°C. Samples were incubated with primary antibody (1/400) for 12 hours at 21°C. A CY3^® conjugated donkey anti-rabbit polyclonal was used as the secondary antibody at 1/200.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Anti-vimentin (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547) staining in E17 rat cheek sections using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/2000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

  Image courtesy of Mr Carl Hobbs, Kings College London.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Formalin-fixed, paraffin-embedded human AD brain tissue stained for vimentin using Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at 1/2000 dilution in immunohistochemical analysis.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Anti-vimentin (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547) staining in adult mouse brain (the dentate gyrus region of the hippocampus) using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/2000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

  Image courtesy of Mr Carl Hobbs, Kings College London.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Anti-vimentin (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547) staining in human ovarian cancer tissue using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/2000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

  Image courtesy of Mr Carl Hobbs, Kings College London.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  This image is courtesy of an anonymous customer review.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 staining Vimentin in rat skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% buffered normal formalin and blocked with 5% serum for 60 minutes at 21°C; antigen retrieval was by heat mediation in a 10mM Sodium citrate buffer. Samples were incubated with primary antibody (1/200 in blocking buffer) for 12 hours at 4°C. A Cy3^®-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Flow Cytometry (Intracellular) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Vimentin flow cytometry staining of HeLa cells using rabbit anti-Vimentin antibody
  
  Overlay histogram showing HeLa cells fixed in 2% PFA and stained with purified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at a dilution of 1 in 50 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Vimentin immunofluorescence staining of HeLa cells using rabbit anti-Vimentin antibody
  
  Immunofluorescence staining of HeLa (human epithelial cell line from cervix adenocarcinoma) cells with purified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at a working dilution of 1/250 counter-stained with DAPI. The secondary antibody was Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 a mouse anti-tubulin antibody (1/1000) was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) 1/1000 shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1 purified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 was used at a dilution of 1/500 followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2 Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at a dilution of 1/400.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Vimentin immunohistochemistry staining of mouse kidney using rabbit anti-Vimentin antibody
  
  Immunohistochemical staining of paraffin embedded mouse kidney with purified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at a working dilution of 1/250. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Vimentin immunofluorescence staining of HeLa cells using rabbit anti-Vimentin antibody
  
  Unpurified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 staining Vimentin in HeLa cells. The cells were fixed with 100% methanol (5 min) permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at a working concentration of 5μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 594 shown in red) at 1/250 dilution overnight at +4°C followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Vimentin immunofluorescence staining of HeLa cells using rabbit anti-Vimentin antibody
  
  Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 staining Vimentin in HeLa (human epithelial cell line from cervix adenocarcinoma) cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at 5μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1μg/ml overnight at +4°C followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody at 2 μg/ml (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

  Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Flow Cytometry (Intracellular) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Overlay histogram showing HeLa cells stained with unpurified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 , 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton X-100 used under the same conditions.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded human cervical carcinoma tissue sections labeling Vimentin with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547.
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded human kidney tissue sections labeling Vimentin with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Vimentin immunohistochemistry staining of human colon using rabbit anti-Vimentin antibody
  
  Fluorescent immunohistochemical analysis of paraffin-embedded human normal colon tissue using unpurified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547. Green- Vimentin red-PI

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Vimentin immunohistochemistry staining of human kidney using rabbit anti-Vimentin antibody
  
  Fluorescent immunohistochemical analysis of paraffin-embedded human normal kidney tissue using unpurified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547. Green- Vimentin red-PI.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  OI-RD Scanning - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  This ICC data was generated using the same anti-Vimentin antibody clone, EPR3776, in a different buffer formulation (cat# Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).
  

  Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 staining Vimentin in wild-type HAP1 cells (top panel) and VIM knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at 0.5μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  This IHC data was generated using the same anti-Vimentin antibody clone, EPR3776, in a different buffer formulation (cat# Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).
  

  IHC image of unpurified Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 staining Vimentin in human breast adenocarcinoma formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

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  Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Vimentin immunofluorescence staining of VIM KO cells using rabbit anti-Vimentin antibody
  
  Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 staining VIM in wild-type HeLa cells with negative expression in VIM knockout HeLa cells. The cells were fixed with 4% formaldehyde (10 min) permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at 2 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488) pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119 Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 647) pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.This product also work with 100% methanol (5 min) fixation under the same testing conditions.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547).

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  Multiplex immunohistochemistry - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Immunofluorescence analysis of human rectum adenocarcinoma labelled with ab193555. Image was acquired using Cell DIVE Multiplexed Imaging Solution (Leica Microsystems).

  Formalin fixed paraffin embedded (FFPE) sections were incubated with ab193555 at 1/100 dilution, for 1h at room temperature. Nuclear DNA was labelled with DAPI (Red). Deparaffinization, antigen retrieval, biomarker labeling and imaging was performed following Cell DIVE recommended standard protocol. Cell DIVE automatically processed the image to perform autofluorescence removal, distortion correction, blank glass subtraction, flat-field correction, and stitching.

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  Multiplex immunohistochemistry - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue.
  
  Panel A: Merged staining of anti-Vimentin (ab193555; red; Opal™690), anti-CYP11A1 (Anti-CYP11A1 antibody [EPR24868-86] ab272494; cyan; Opal™520) and anti-DDX4 / MVH (Anti-DDX4 / MVH antibody [EPR24148-58] ab270534; green; Opal™570) on human testis.
  
  Panel B: Anti-CYP11A1 stained on Leydig cells.
  
  Panel C: Anti-DDX4 / MVH stained on all spermatogenic cell types.
  
  Panel D: Anti-Vimentin stained on Sertoli cells and fibroblasts.
  
  Key protocol steps: The section was incubated in three rounds of staining: in the order of ab193555 (1:2000 dilution) and Anti-CYP11A1 antibody [EPR24868-86] ab272494 (1:10000 dilution) for 30 mins, then Anti-DDX4 / MVH antibody [EPR24148-58] ab270534 (1:2000 dlilution) for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
  
  DAPI was used as a nuclear counter stain. Opal Polymer HRP Ms + Rb was used as a secondary.
  
  Antigen retrieval: Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)

  This data was developed using Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547, the same antibody clone in a different buffer formulation.
  
  Tissue Microarrays stained for Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker using Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested. The section was incubated with Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547 at 4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) secondary antibody (Goat Anti-Rabbit IgG H&L (HRP polymer) ab214880).
  
  Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).