Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

This data was developed using the same antibody clone in a different buffer formulation (ab52866).

Immunohistochemical analysis of formalin fixed paraffin embedded human kidney staining alpha Tubulin with ab52866 at a concentration of 0.01 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab52866 anti- alpha Tubulin [EP1332Y] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

Immunohistochemistry analysis of paraffin-embedded Human breast cancer labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on cancer cells shown. Secondary antibody ab97051 Goat Anti-Rabbit IgG H&L (HRP) used at a 1/500 dilution. Counter stained with Hematoxylin.

Inset image : negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

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

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

AJAP1 co-localizes with microtubules in HUVECs

The association of AJAP1 with microtubules in HUVECs is lost upon microtubule destruction. Treatment with 12.5 µM nocodazole for 24 h shows destruction of the microtubule network and loss of AJAP1 tubular localization. For a negative control, HUVECs are treated with DMSO for 24 h. Cell nuclei were counterstained with DAPI (cyan). Microscope : Zeiss LSM 780; objective lens : 63×/1.40 oil; scale bar : 25 µm.

Incubated overnight at 4°C with ab52866.

(From Figure 3E of Hotte et al)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

Image from Hotte K. et al Biol Open. 2017 Jun 15;6(6):723-731. doi: 10.1242/bio.022335.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

This ICC data was generated using the same anti-alpha Tubulin antibody clone, EP1332Y, in a different buffer formulation (ab52866).

Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling alpha Tubulin with ab52866 at 1/500 dilution. The cells were permeabilised with 0.1% Triton X-100. Anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows microtubules staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution (red).
The negative controls are as follows :
1. ab52866 at 1/500 dilution followed by anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution.

• ICC/IF

AbReview20358****

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

ab52866 staining alpha Tubulin in 293 Human embryonic kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 2 hours at 23°C. Samples were incubated with primary antibody (1/200 in 0.5% saponin) for 2 hours at 23°C. An Alexa Fluor^®555-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were counterstained with DAPI.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

This image is courtesy of an anonymous Abreview.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

Clone EP1332Y (ab216650) has been successfully conjugated by Abcam. This image was generated using Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Alexa Fluor® 488). Please refer to ab185031 for protocol details.

ab185031 staining alpha-Tubulin in HeLa 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 ab185031 at a working dilution of 1 in 100 overnight at +4°C (shown in green). Alexa Fluor^® 350 WGA was used at a 1/200 dilution and incubated for 1h with the cells, to label plasma membranes (shown in blue). Nuclear DNA was labelled in red with 1.25 μM DRAQ5™ (ab108410).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

Clone EP1332Y (ab216650) has been successfully conjugated by Abcam. This image was generated using Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (PE). Please refer to ab208752 for protocol details.

ab208752 staining alpha Tubulin in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab208752 at 1/500 dilution (Pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

Clone EP1332Y (ab216650) has been successfully conjugated by Abcam. This image was generated using Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Alexa Fluor® 647). Please refer to ab190573 for protocol details.

ab190573 staining alpha Tubulin in HeLa 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 ab190573 at a working dilution of 1 in 100 (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

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

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

Intracellular Flow Cytometry analysis of 2% paraformaldehyde fixed HepG2 (human liver hepatocellular carcinoma cell line) cells labeling alpha Tubulin with ab52866 at 1/130 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Rat kidney tubule and weak on glomerulus shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.

Inset image : negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

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-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

Immunohistochemistry analysis of paraffin-embedded Mouse kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Mouse kidney tubule shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.

Inset image : negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

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-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (AB216650)

This IHC data was generated using the same anti-alpha Tubulin antibody clone, EP1332Y, in a different buffer formulation (cat# ab52866).

Immunohistochemistry analysis of paraffin-embedded Pig kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Pig kidney tubule and weak on glomerulus shown. Anti-Rabbit HRP (ab97051) used at a 1/100 dilution. Counter stained with Hematoxylin.

Inset image : negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/100 dilution.

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