Anti-GGT1/GGT + GGT3 antibody [EPR28715-82]

8 product images

• 

  Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768)

  Amongst human GGT homologous proteins, this antibody reacts with GGT1 and GGT3 but not GGT2.

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate 7 at 7 µg with 5% NFDM/TBST

  Lane 2: 293T cells transfected with a human GGT1 expression vector containing a myc-His-tag®, whole cell lysate at 7 µg with 5% NFDM/TBST

  Lane 3: 293T cells transfected with a human GGT2 expression vector containing a myc-His-tag®, whole cell lysate at 7 µg with 5% NFDM/TBST

  Lane 4: 293T cells transfected with a human GGT3 expression vector containing a myc-His-tag®, whole cell lysate at 7 µg with 5% NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 60-75 kDa, 36 kDa, 60 kDa, 75 kDa

  Exposure time: 37s

• 

  Immunoprecipitation - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768)

  GGT1/GGT + GGT3 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab316768 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316768 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

  Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
  
  Lane 2: ab316768 IP in HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
  
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab316768 in HepG2 whole cell lysate

  All lanes: Immunoprecipitation - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768) at 1/30 dilution

  All lanes: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 76s

• 

  Immunocytochemistry/ Immunofluorescence - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling GGT1/GGT + GGT3 with ab316768 at 1/50 (9.98 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

  Confocal image showing mainly membranous staining in HepG2 cells.
  
  Low expression: HEK-293 (PMID: 23682772).
  
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768)

  Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling GGT1/GGT + GGT3 with ab316768 at 1/100 (4.99 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on bile canaliculi of human liver (PMID: 8813074).
  
  The section was incubated with ab316768 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
  
  Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768)

  Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling GGT1/GGT + GGT3 with ab316768 at 1/100 (4.99 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on proximal tubules of human kidney (PMID: 8813074, 32474759).
  
  The section was incubated with ab316768 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768)

  Low expression: skeletal muscle (PMID: 7689219).

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  All lanes: Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768) at 1/1000 dilution

  Lane 1: Human kidney tissue lysate at 20 µg with 5% NFDM/TBST

  Lane 2: Human liver tissue lysate at 20 µg with 5% NFDM/TBST

  Lane 3: Human skeletal muscle tissue lysate at 20 µg with 5% NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 60-75 kDa, 36 kDa

  Exposure time: 37s

• 

  Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768)

  Low expression: HEK-293 (PMID: 23682772).

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  All lanes: Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768) at 1/1000 dilution

  Lane 1: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 2: HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 3: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 4: Untreated HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 5: HepG2 whole cell lysate treated with Protein Deglycosylation Mix II, at 20 µg with 5% NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 60-75 kDa, 36 kDa

  Exposure time: 81s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (ab316768)

  Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling GGT1/GGT + GGT3 with ab316768 at 1/100 (4.99 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Negative control: no staining on human cardiac muscle.
  
  The section was incubated with ab316768 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
  
  Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.