Anti-Smg1 antibody

• ICC/IF

AbReview6772****

Immunocytochemistry/ Immunofluorescence - Anti-Smg1 antibody (AB30916)

ab30916 (1/200) staining Smg1 by ICC/IF in assynchronous HeLa Cells (green). Cells were countersatined with DAPI (red) in order to show the nucleus. Secondary antibody : Goat anti-Rabbit conjugated to Cy3®. Please refer to abreview for further details.

This image is courtesy of an Abreview submitted by Dr Kirk McManus

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Smg1 antibody (AB30916)

ICC/IF image of ab30916 stained HepG2 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 (ab30916, 5µg/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) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml

• WB

Project

Western blot - Anti-Smg1 antibody (AB30916)

The band at 340kDa is likely to represent isoform 3 of Smg1. The immunogen is present in all three isoforms of Smg1.

All lanes:

Western blot - Anti-Smg1 antibody (ab30916) at 1 µg/mL

Lane 1:

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

Lane 2:

Western blot - Jurkat whole cell lysate (<a href='/en-us/products/cell-lysates/jurkat-whole-cell-lysate-ab7899'>ab7899</a>) at 10 µg

Lane 3:

Western blot - A-431 whole cell lysate (<a href='/en-us/products/cell-lysates/a-431-whole-cell-lysate-ab7909'>ab7909</a>) at 20 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

Predicted band size: 411 kDa

Observed band size: 340 kDa

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