Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68193).

Flow cytometry overlay histogram showing wild-type HeLa (green line) and TUBB3 knockout HeLa stained with ab68193 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab68193) (1x 106 in 100μl at 0.04 μg/ml (1/49250)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type HeLa - black line, TUBB3 knockout HeLa - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling beta III Tubulin with Purified ab68193 at 1 : 8000 dilution (0.014 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.

ab68193 staining beta III Tubulin in wild-type HAP1 cells (top panel) and TUBB3 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), 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 with ab68193 at 1μg/ml concentration and ab7291, a mouse anti-tubulin antibody, at 1μg/ml overnight at +4°C. This is followed by a further incubation at room temperature for 1h with a goat anti-rabbit IgG Alexa Fluor® 488 (ab150081) at 2μg/ml (shown in green) and a goat anti-mouse IgG Alexa Fluor® 647 (ab150119) at 2μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.

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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling beta III Tubulin with Purified ab68193 at 1 : 20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.

Overlay histogram showing HeLa cells stained with ab68193 (red line). The cells were fixed with 80% methanol (5 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 (ab68193, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Anti-rabbit Alexa Fluor® 488 (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 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.

ab68193 staining beta III Tubulin in Neuro-2a 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 ab68193 at 1μg/ml and ab7291 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls : 1– Rabbit primary 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of Neuro-2a(Mouse neuroblastoma neuroblast) cells labeling beta III Tubulin with Purified ab68193 at 1 : 50 dilution (2.2 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue sections labeling beta III Tubulin with Purified ab68193 at 1 : 8000 dilution (0.014 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling beta III Tubulin with Purified ab68193 at 1 : 8000 dilution (0.014 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• WB

Unknown

Western blot - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-beta III Tubulin antibody [EPR1568Y] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/beta-iii-tubulin-antibody-epr1568y-neuronal-marker-ab68193'>ab68193</a>) at 1/50000 dilution

Lane 1:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Human cerebellum lysate at 20 µg

Lane 3:

Mouse brain lysate at 20 µg

Lane 4:

Rat brain lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

• WB

Unknown

Western blot - Anti-beta III Tubulin antibody [EPR1568Y] - BSA and Azide free (AB247375)

This data was developed using ab68193, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-beta III Tubulin antibody [EPR1568Y] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/beta-iii-tubulin-antibody-epr1568y-neuronal-marker-ab68193'>ab68193</a>) at 1/50000 dilution

All lanes:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false