Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free

10 product images

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  Flow Cytometry (Intracellular) - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  Overlay histogram showing HeLa cells stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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-alpha Tubulin antibody [DM1A] - Loading Control ab7291, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was an anti mouse DyLight® 488 (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291).

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  Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 staining alpha Tubulin in SV40LT-SMC 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 Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at a working concentration of 0.5μg/ml and Alexa Fluor® 647 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab190573, Rabbit monoclonal [EP1332Y] to alpha Tubulin (Alexa Fluor® 647, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
  

  This product also gave a positive signal in 100% methanol (5 min) fixed SV40 cells under the same testing conditions.

  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, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291)

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  Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 staining alpha Tubulin in NIH3T3 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-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1μl/ml and Anti-beta Tubulin antibody - Loading Control ab6046 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 μg/ml (shown in green) and anti-rabbit AlexaFluor® 594 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088) 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, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  IHC image of Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 staining alpha Tubulin in human colon 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-alpha Tubulin antibody [DM1A] - Loading Control ab7291, 5ug/ml working concentration, 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 secondary only 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
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291)

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  Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 staining alpha-Tubulin in Caco-2 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-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1μg/ml and Anti-beta Tubulin antibody - Loading Control ab6046 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 μg/ml (shown in green) and anti-rabbit AlexaFluor® 594 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088) 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, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291).

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  Western blot - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  Merged signal (red and green). Green - Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 observed at 52 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, observed at 38 kDa.
  

  All samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/1,000 and 1/20,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291)

  All lanes: Western blot - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution

  Lane 1: HeLa cell lysate at 20 µg

  Lane 2: PC12 cell lysate at 20 µg

  Lane 3: SV40LT-SMC cell lysate at 20 µg

  Lane 4: NIH/3T3 cell lysate at 20 µg

  Lane 5: Rat liver tissue lysate at 20 µg

  Lane 6: Rat heart tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 50 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  IHC image of Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 staining alpha Tubulin in rat colon 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-alpha Tubulin antibody [DM1A] - Loading Control ab7291, 0.5ug/ml working concentration, 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 secondary only 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.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291).

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  Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  FABP4 (green) was detected using FABP4 primary antibody (Anti-FABP4 antibody [EPR3579] ab92501; diluted 1/1000). Alpha tubulin (red) was detected using the mouse monoclonal (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) antibody. Cells were imaged by confocal microscopy, using z-stack for adipocyte-like cells.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291).

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  Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  ICC/IF image of Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, 1μg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291).

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  Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

  Immunofluorescent imaging of human cells (U2OS) with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 reveals a delicate network of alpha-tubulin (green) located exclusively in the cytoplasm. The nucleus is stained blue.
  

  IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour. Primary antibody used at 1/200 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes. All blocking and incubation steps carried out at 37 degrees.

  Unfortunately, due to size constraints for images on our website, we are unable to show the full uncompressed picture.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291).