Anti-TACC3 antibody [EPR7756] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TACC3 antibody [EPR7756] - BSA and Azide free (AB224525)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TACC3 with purified ab134154 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

Control : primary antibody (1/200) and secondary antibody, ab150113, an Alexa Fluor^® 488-conjugated goat anti-mouse IgG (1/500).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134154).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-TACC3 antibody [EPR7756] - BSA and Azide free (AB224525)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TACC3 with unpurified ab134154 at 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134154).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TACC3 antibody [EPR7756] - BSA and Azide free (AB224525)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling TACC3 with unpurified ab134154 at 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134154).

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-TACC3 antibody [EPR7756] - BSA and Azide free (AB224525)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling TACC3 with unpurified ab134154 at 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134154).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TACC3 antibody [EPR7756] - BSA and Azide free (AB224525)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling TACC3 with purified ab134154 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134154).

• WB

Lab

Western blot - Anti-TACC3 antibody [EPR7756] - BSA and Azide free (AB224525)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134154).

Blocking and diluting buffer and concentration : 5% NFDM/TBST

In Western blot, ab134154 was shown to bind specifically to TACC3. Target of interest was observed at 140 kDa in wild-type 293T cell lysates (lane 1) with no signal observed at this size in TACC3 knockout cell line (lane 2) (lane 2, knockout cell line ab266316).

The bands beneath the target band (140 kDa) are likely to be degraded target fragments.

ab181602 was used as a GAPDH loading control at 1/200000 dilution.

All lanes:

Western blot - Anti-TACC3 antibody [EPR7756] (<a href='/en-us/products/primary-antibodies/tacc3-antibody-epr7756-ab134154'>ab134154</a>) at 1/1000 dilution

Lane 1:

Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TACC3 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-tacc3-knockout-hek-293t-cell-line-ab266316'>ab266316</a>) at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 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 1/100000 dilution

Predicted band size: 90 kDa

Observed band size: 140 kDa,37 kDa

false

Exposure time: 81s

• WB

Lab

Western blot - Anti-TACC3 antibody [EPR7756] - BSA and Azide free (AB224525)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134154).

Blocking and diluting buffer and concentration : 5% NFDM/TBST

To minimize protein degradation, tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

All lanes:

Western blot - Anti-TACC3 antibody [EPR7756] (<a href='/en-us/products/primary-antibodies/tacc3-antibody-epr7756-ab134154'>ab134154</a>) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Rat testis tissue lysate at 20 µg

Lane 3:

Rat spleen tissue lysate at 20 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 1/100000 dilution

Predicted band size: 90 kDa

Observed band size: 140 kDa

false

Exposure time: 125s