Anti-O-Linked N-Acetylglucosamine 抗体 [RL2]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (AB2739)

ab2739 stained in MCF7 cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab2739 at 5μg/ml and ab6046 (Rabbit polyclonal to beta Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150080 (pseudo-colored red) and ab150117 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43μM for 1hour at room temperature.

• WB

Lab

Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (AB2739)

Jurkat cells were treated with either 50 uM PugNAc (ab144670) or 4 mM glucosamine + 50 uM PugNAc (ab144670) for three hours prior to harvest to stimulate O-linked glycosylation. The expected increase in glycosylation is observed in the treated lanes 2 & 4.

All lanes:

Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739) at 1 µg/mL

Lanes 1 and 3:

Jurkat cells treated with 0 uM PugNAc at 20 µg

Lane 2:

Jurkat cells treated with 50 uM PugNAc (3 hours) at 20 µg

Lane 4:

Jurkat cells treated with 4 mM glucosamine and 50 uM PugNAc (3 hours) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/50000 dilution

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Exposure time: 12min

• WB

Project

Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (AB2739)

The antibody was tested against the immunogen (isolated rat liver nuclear envelopes, which contain 8 O-linked glycoproteins in the nuclear pore complex).

All lanes:

Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739) at 1 µg/mL

All lanes:

Rat Liver Nuclear Envelope at 10 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

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Exposure time: 1min

• WB

AbReview42315****

Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (AB2739)

All lanes:

Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739) at 1/3000 dilution

Lane 1:

Human neuroblastoma (SH-SY5Y) whole cell lysate - treated with 50µM z-Pugnac for 24 hours at 20 µg

Lane 2:

Human neuroblastoma (SH-SY5Y) whole cell lysate - untreated at 20 µg

Secondary

All lanes:

HRP-conjugated horse anti-mouse IgG polyclonal

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Exposure time: 30s

This image is courtesy of an anonymous Abreview