Anti-Sumo 3 antibody [1H9L17]

• Flow Cyt

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Flow Cytometry - Anti-Sumo 3 antibody [1H9L17] (AB203570)

Flow cytometric analysis of fixed and permealized Jurkat cells labeling Sumo 3 using ab203570 at 0.5 μg/test (Black) followed by secondary detection with Alexa Fluor® 488 Goat anti-Rabbit. Cells incubated without primary antibody (grey) for control. Preincubation with immunizing peptide decreased the staining (red).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 3 antibody [1H9L17] (AB203570)

Immunohistochemical analysis of formalin fixed, paraffin embedded Human colon carcinoma tissue labeling Sumo 3 using ab203570 at 5 μg/mL, followed by DAB detection.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Sumo 3 antibody [1H9L17] (AB203570)

Immunofluorescence analysis of 70% confluent log phase MCF7 cell labelled for SMT3/ SUMO3 using ab207570 at 1μg/mL. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ab203570 in 1% BSA and incubated for 3 hours at room temperature and then labeled with secondary antibody for 45 minutes at room temperature (Panel A : green). Nuclei (Panel B : blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel C : red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin(1/300). Panel D is a merged image showing Nuclear localization. Panel E is a no primary antibody control. The images were captured at 60X magnification.

• WB

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Western blot - Anti-Sumo 3 antibody [1H9L17] (AB203570)

Western blot analysis of SUMO-3 expression in Mouse Esophagus, Mouse Skin, Mouse Skeletal muscle, and human cell lines Jurkat and HepG2. Used Novex® NuPAGE®4-12 % Bis-Tris gel, XCell SureLock™ Electrophoresis System, Novex® Sharp Pre-Stained Protein Standard, and iBlot® Dry Blotting System. Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature.

SUMO-3 was detected at ~ 20 kDa using ab203570 in 5 % skim milk at 4°C overnight on a rocking platform. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate.

All lanes:

Western blot - Anti-Sumo 3 antibody [1H9L17] (ab203570) at 0.1 µg/mL

Lane 1:

Mouse Esophagus at 20 µg

Lane 2:

Mouse skin at 20 µg

Lane 3:

Mouse Skeletal muscle at 20 µg

Lane 4:

Jurkat at 20 µg

Lane 5:

Hep G2 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG - HRP Secondary Antibody at 0.02 µg/mL

Predicted band size: 11 kDa

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• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Sumo 3 antibody [1H9L17] (AB203570)

Immunofluorescent analysis of U2OS cells labeling Sumo 3 using ab203570 at 5 μg/mL (left image). Right image : sample with immunogenic phosphopeptide. Alexa Fluor® 488 Goat anti-Rabbit at 1/1000 dilution was used as secondary antibody.