Anti-TRIM33 antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TRIM33 antibody (AB47062)

ab47062 staining TRIM33 in MCF7 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-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 ab47062 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Unknown

Western blot - Anti-TRIM33 antibody (AB47062)

All lanes:

Western blot - Anti-TRIM33 antibody (ab47062) at 1 µg/mL

Lane 1:

SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 3:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Lane 4:

SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

Lane 5:

K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 122 kDa

Observed band size: 150 kDa,220 kDa

true

Exposure time: 30s

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-TRIM33 antibody (AB47062)

ICC/IF image of ab47062 stained mouse embryonic stem cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47062, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).

• WB

Project

Western blot - Anti-TRIM33 antibody (AB47062)

The 150 kDa band observed is comparable to molecular weights seen with other commercially available antibodies to TRIM33.

All lanes:

Western blot - Anti-TRIM33 antibody (ab47062) at 1 µg/mL

Lane 1:

F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate (<a href='/en-us/products/unavailable/f9-mouse-embryonic-carcinoma-cell-line-whole-cell-lysate-ab27193'>ab27193</a>) at 10 µg

Lane 2:

E14tG2a (Mouse embryonic stem cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 122 kDa

Observed band size: 150 kDa,220 kDa

false

• WB

Lab

Western blot - Anti-TRIM33 antibody (AB47062)

Gel type : TA Blocking buffer : 1% milk block

All lanes:

Western blot - Anti-TRIM33 antibody (ab47062) at 1 µg/mL

Lane 1:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

A431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

Mouse embryonic stem cell lysate at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 122 kDa

Observed band size: 150 kDa

false

Exposure time: 8min