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

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

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  Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)
  All lanes : Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739) at 1 µg/ml
  
  Lane 1 : Jurkat cells treated with 0 uM PugNAc
  Lane 2 : Jurkat cells treated with 50 uM PugNAc (3 hours)
  Lane 3 : Jurkat cells treated with 0 uM PugNAc
  Lane 4 : Jurkat cells treated with 4 mM glucosamine and 50 uM PugNAc (3 hours)
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Exposure time : 12 minutes

  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. 

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  Immunocytochemistry/ Immunofluorescence - O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)
  ICC/IF image of ab2739 stained HeLa 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 (ab2739, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Mouse 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) HEK293, HepG2, and MCF-7 cells at 10µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, and MCF-7 cells at 5µg/ml.
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  Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)This image is courtesy of an anonymous Abreview
  All lanes : 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
  Lane 2 : Human neuroblastoma (SH-SY5Y) whole cell lysate - untreated
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  HRP-conjugated horse anti-mouse IgG polyclonal
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Exposure time : 30 seconds

  This image is courtesy of an anonymous Abreview

  See Abreview

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  Western blot - O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)
  Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739) at 1 µg/ml + Rat Liver Nuclear Envelope at 10 µg
  
  Secondary
  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 : 1 minuteThe antibody was tested against the immunogen (isolated rat liver nuclear envelopes, which contain 8 O-linked glycoproteins in the nuclear pore complex).