Anti-beta Tubulin antibody [EPR1330] - Loading Control

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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 >5000 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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.

• WB

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Western blot - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

All lanes:

Western blot - Anti-beta Tubulin antibody [EPR1330] - Loading Control (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

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• ICC/IF

AbReview40712****

Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

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.

This image is courtesy of an Abreview submitted by Kirk McManus

• WB

Unknown

Western blot - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

Blocking/Diluting buffer 5% NFDM /TBST

All lanes:

Western blot - Anti-beta Tubulin antibody [EPR1330] - Loading Control (ab108342) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma) whole cell lysate at 20 µg

Lane 2:

C6 (rat glioma) whole cell lysate at 20 µg

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

false

• WB

Unknown

Western blot - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

Blocking/Diluting buffer 5% NFDM /TBST

All lanes:

Western blot - Anti-beta Tubulin antibody [EPR1330] - Loading Control (ab108342) at 1/5000 dilution

Lane 1:

Hela (human cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Human fetal brain tissue lysate at 20 µg

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

false

• WB

CiteAb

Western blot - Anti-beta Tubulin antibody [EPR1330] - Loading Control (AB108342)

beta Tubulin western blot using anti-beta Tubulin antibody [EPR1330] - Microtubule Marker ab108342. Publication image and figure legend from Hur, J. H., Park, S. Y., et al., 2016, Sci Rep, PubMed 27976696.

ab108342 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab108342 please see the product overview.

Pld1 expression is decreased in HFD-induced hepatic steatosis.(a) C57BL/6 mice were fed either RC or a HFD for 4 weeks (n = 10 per group) starting at the age of 13 weeks and fasted overnight prior to collection of the liver. mRNA was isolated from the liver for qRT-PCR. (b) Mice were maintained as in (a) and the lysates were analyzed by western blot. Western blot results were analyzed by densitometry to obtain the relative ratio of either PLD1 or PLD2 to tubulin. The data are presented as means ± SE or representative blots from 3 to 5 independent experiments. *p < 0.05 versus the RC group. Abbreviations : HFD, high-fat diet; PLD2, phospholipase D2; RC, regular chow; SE, standard error; qRT-PCR quantitative real-time polymerase chain reaction.

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