Western blot - Vimentin antibody - Neural Stem Cell Marker (ab45939)

All lanes : Anti-Vimentin antibody - Neural Stem Cell Marker (ab45939) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
Lane 4 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 54 kDa
Observed band size : 54 kDa

Western blot - Vimentin antibody - Neural Stem Cell Marker (ab45939)

All lanes : Anti-Vimentin antibody - Neural Stem Cell Marker (ab45939) at 1 µg/ml

Lane 1 : Vimentin protein (Human) (ab73843) at 0.1 µg
Lane 2 : Vimentin protein (Human) (ab73843) at 0.01 µg

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Observed band size : 54 kDa


Exposure time : 10 seconds

Ab45939 recognizes full length recombinant Human vimentin (ab73843) which has an expected molecular weight of 54 kDa.

Immunocytochemistry/ Immunofluorescence - Vimentin antibody - Neural Stem Cell Marker (ab45939)

ICC/IF image of ab45939 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab45939, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Vimentin antibody - Neural Stem Cell Marker (ab45939)

Immunohistochemical detection (formaldehyde/paraffin-embedded sections) of vimentin protein using vimentin antibody - Neural Stem Cell Marker (ab45939) on human ovarian carcinoma sections. Antigen retrieval step:Heat mediated. Blocking step: 1% BSA for 10 mins at RTºC. Ab45939 was used at 1/700 for 1 hour. Secondary Antibody: Biotin-conjugated anti rabbit IG (1/300).

Carl Hobbs, CARD, KCL, UK

Immunohistochemistry (Frozen sections) - Vimentin antibody - Neural Stem Cell Marker (ab45939)

ab45939 staining Vimentin in human squamous cell carcinoma tissue sections by IHC-Fr (Frozen sections). Tissue samples were fixed with formaldehyde and blocked with 4% milk for 20 minutes at 22ºC. The sample was incubated with primary antibody (1/50 in milk) at 4ºC for 18 hours. A Biotin-conjugated Goat polyclonal to rabbit IgG (1/200) was used as secondary antibody.

This image is courtesy of an anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Vimentin antibody - Neural Stem Cell Marker (ab45939)

ab45939 staining Vimentin in murine kidney tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 10% serum for 30 minutes at room temperature followed by incubation with the primary antibody at a 1/2100 dilution for 18 hours at 4ºC. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/200 dilution.

Image courtesy of Mike Forbes by Abreview.

Immunocytochemistry/ Immunofluorescence - Vimentin antibody - Neural Stem Cell Marker (ab45939)

ICC/IF image of ab45939 stained Hela cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45939, 1µg/ml) overnight at +4ºC. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.