Anti-Src antibody (ab47405) at 1/2000 dilution + HeLa cells

Predicted band size: 60 kDa

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Src antibody, in the presence and absence of blocking peptide.

All lanes : Anti-Src antibody (ab47405) at 1/500 dilution

Lane 1 : HEK-293 cell extracts
Lane 2 : HEK-293 cell extracts with Blocking peptide

Predicted band size: 60 kDa
Observed band size: 60 kDa



Western blot analysis of extracts from 293 cells using Src antibody in the presence and absence of blocking peptide. Western blot analysis of extracts from HEK-293 cells using Src antibody at 1/500 in the presence and absence of blocking peptide.

ab47405 staining Src in human HEK-293 cells by Immunocytochemistry/ Immunofluorescence. The cells were paraformaldehyde fixed, permeabilised in 0.5% Triton X-100 and then blocked using 5% serum for 20 minutes at 25°C. Samples were then incubated with primary antibody at 1/1000 for 16 hours at 4°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/5000 dilution. DAPI was used to stain the cell nuclei (blue).

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ab47405 staining Src in Pig retinal pigment epithelium primary cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilzed with 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/1000 in 0.1% TX100, 1% goat serum, 1XPBS) for 16 hours at 4°C. An Alexa Fluor^®488-conjugated goat anti-rabbit IgG polyclonal (1/5000) was used as the secondary antibody. Nuclei were counterstained with DAPI.

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ICC/IF image of ab47405 stained MCF7 cells (ab3871). 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 (ab47405, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) used at a 1/1000 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.

Anti-Src antibody (ab47405) at 1/500 dilution + Recombinant human Src protein (ab79635) at 0.001 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 60 kDa


Exposure time: 10 seconds

Anti-Src antibody (ab47405) at 1/1000 dilution + whole cell lysate prepared from mouse primary astrocytes at 15 µg

Secondary
Goat abti-rabbit IgG conjugated to HRP at 1/5000 dilution

Performed under reducing conditions.

Predicted band size: 60 kDa
Observed band size: 60 kDa


Exposure time: 5 seconds


Primary antibody incubated for 1 hour at 25°C.Blocked with 3& BSA for 1 hour at 25°C.Gel run under denaturing conditions.Detection method: ImmunoStar.

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