All lanes : Anti-beta Tubulin antibody [EPR1330] (ab108342) at 1/5000 dilution

Lane 1 : Hela (human cervix adenocarcinoma) whole cell lysate
Lane 2 : Human fetal brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 49 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?



Blocking/Diluting buffer 5% NFDM /TBST

Immunohistochemical analysis of paraffin-embedded human kidney tissue sections labelling beta Tubulin with purified ab108342 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. Sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

Immunocytochemistry/Immunofluorescence staining of HeLa cells labelling beta Tubulin with purified ab108342 at a working dilution of 1/500. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. DAPI was used as nuclear counterstain (bottom left hand panel). The cells were fixed in 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, rabbit primary antibody was used followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120). For negative control 2, ab7291 (mouse anti-tubulin) was used followed by an Alexa Fluor® 488 goat anti-rabbit secondary (ab150077).

Overlay histogram showing 4% paraformaldehyde fixed Hela (human cervix adenocarcinoma) cells labelling beta Tubulin with purified ab108342 at dilution of 1/20. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG at dilution of 1/2000. A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black line). The blue line shows cells without incubation with primary antibody and secondary antibody. 

All lanes : Anti-beta Tubulin antibody [EPR1330] (ab108342) at 1/1000 dilution

Lane 1 : Neuro-2a (mouse neuroblastoma) whole cell lysate
Lane 2 : C6 (rat glioma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 49 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?



Blocking/Diluting buffer 5% NFDM /TBST

Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue sections labelling beta Tubulin with purified ab108342 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. Sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

All lanes : Anti-beta Tubulin antibody [EPR1330] (ab108342) at 1/1000 dilution

Lane 1 : MCF-7 cell lysates
Lane 2 : Jurkat cell lysates
Lane 3 : Ramos cell lysates
Lane 4 : HeLa cell lysates

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 49 kDa

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue sections labelling beta Tubulin with purified ab108342 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. Sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

Immunofluorescent staining of HeLa cells using ab108342 at 1/100 dilution.

ab108342 staining beta Tubulin in human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/200 in PBS) for 1 hour at 22°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counterstained with DAPI.

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Overlay histogram showing HeLa cells stained with ab108342 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab108342, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

ab108342 at 1/250 dilution staining beta Tubulin in Human brain tissue by Immunohistochemistry Formalin-fixed, Paraffin-embedded tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab108342 at 1/250 dilution staining beta Tubulin in Human tonsil tissue by Immunohistochemistry Formalin-fixed, Paraffin-embedded tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab108342 showing positive staining in Normal human kidney tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab108342 showing positive staining in human Breast carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.