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

4 product images

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  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 Anti-beta Tubulin antibody - Loading Control ab6046 (Rabbit polyclonal to beta Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 (pseudo-colored red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed 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.

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  Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)

  Jurkat cells were treated with either 50 uM PugNAc (PugNAc, hexosaminidase A and B inhibitor ab144670) or 4 mM glucosamine + 50 uM PugNAc (PugNAc, hexosaminidase A and B inhibitor 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 (Goat Anti-Mouse IgG H&L (HRP) preadsorbed ab97040) at 1/50000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Exposure time: 12min

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  This image is courtesy of an anonymous customer review

  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

  Developed using the ECL technique.

  Performed under reducing conditions.

  Exposure time: 30s

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  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

  Developed using the ECL technique.

  Performed under reducing conditions.

  Exposure time: 1min