Anti-Dishevelled 3 antibody [EP1991Y] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Dishevelled 3 antibody [EP1991Y] - BSA and Azide free (AB284737)

This data was developed using ab76081, the same antibody clone in a different buffer formulation.

Flow cytometry analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) labelling Dishevelled 3 with ab76081 at 1/200 dilution, followed by blocking with 10% goat serum for 1h at room temperature. ab76081 was incubated for 30 min at room temperature (RT) and then Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) was incubated for 30 min at room temperature.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Dishevelled 3 antibody [EP1991Y] - BSA and Azide free (AB284737)

This data was developed using ab76081, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 cells labelling Dishevelled 3 with ab76081 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) at 1/1000 dilution at RT for 45 min. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889) was used to counterstain tubulin at 1/200 dilution. Nucleus were visualized using DAPI.

• WB

Lab

Western blot - Anti-Dishevelled 3 antibody [EP1991Y] - BSA and Azide free (AB284737)

This data was developed using ab76081, the same antibody clone in a different buffer formulation.

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

Exposure time : Lane 1, 20 seconds; Lane 2 and Lane 3, 40 seconds.

Double bands of DVL3 could be observed which was supposed to be caused by phosphorylation supported by literatures (PMID : 31870452, 22647544).

All lanes:

Western blot - Anti-Dishevelled 3 antibody [EP1991Y] (<a href='/en-us/products/primary-antibodies/dishevelled-3-antibody-ep1991y-ab76081'>ab76081</a>) at 1/1000 dilution

Lane 1:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 15 µg

Lane 2:

RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 15 µg

Lane 3:

C6 (Rat glial tumor glial 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

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