Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free

20 product images

• 

  Zhu, Y. et al PLoS Pathog. 2016 May 17;12(5):e1005648. doi: 10.1371/journal.ppat.1005648. eCollection 2016 May Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  GLUT1 and GLUT3 are downregulated in KSHV-infected cells in human KS tumors
  

  Representative illustration of dual immunofluorescence detection of LANA and GLUT1 or in a normal human skin section and a Karposi Sarcoma (KS) tumor section. Tissues were fixed with paraformaldehyhde and paraffin-embedded.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

• 

  Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Blocking buffer and concentration: 5% NFDM/TBST
  

  Diluting buffer and concentration: 5% NFDM/TBST

  All lanes: Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Lane 1: HepG2 (human hepatocellular carcinoma) whole cell lysate at 15 µg

  Lane 2: HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051)

  Predicted band size: 54 kDa

  Observed band size: 40-60 kDa

  Exposure time: 3min

• 

  Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196357).
  
  

  Lanes 1 - 2: Merged signal (red and green). Green - ab196357 observed at 54 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

  ab196357 was shown to recognize in wild-type A549 cells as signal was lost at the expected MW in SLC2A1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SLC2A1 knockout samples were subjected to SDS-PAGE. ab196357 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730) at 1 µg/mL

  Lane 1: Wild-type A549 whole cell lysate at 20 µg

  Lane 2: Western blot - Human SLC2A1 (Glucose Transporter GLUT1) knockout A549 cell line (Human SLC2A1 (Glucose Transporter GLUT1) knockout A549 cell line ab261869) at 20 µg

  Predicted band size: 54 kDa

• 

  Khaom, R. et al PLoS One. 2016 Aug 11;11(8):e0161163. doi: 10.1371/journal.pone.0161163. eCollection 2016 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Immunohistochemical expression of Glut1 in normal tongue epithelium and tongue cancer. Expression was greatest in lymphocytes (arrows in left upper and lower panels). In the normal oral epithelium, Glut1 was weakly expressed in the basal and spinous cells (left upper panel). In OSCC, Glut1 was upregulated, showing a level of expression comparable with lymphocytes (left and right lower panels). Scale bar, 100 μm.

  Note: Glut1 = SLC2A (alternative names for the same target).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Immunofluorescence staining of HepG2 cells with purified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

• 

  Flow Cytometry (Intracellular) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 at a dilution of 1/40 (red line). The secondary antibody used was Alexa Fluorr^® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Immunohistochemical staining of paraffin embedded human lung carcinoma with purified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 at a working dilution of 1/500. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 at a working dilution of 1/500. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

• 

  Flow Cytometry (Intracellular) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Overlay histogram showing HeLa cells stained with unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 at 1/250 dilution staining Glucose Transporter GLUT1 in Paraffin-embedded human colonic adenocarcinoma tissue by Immunohistochemistry.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 showing positive staining in normal liver tissue.
  
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 at 1/250 dilution staining Glucose Transporter GLUT1 in Paraffin-embedded human cervical carcinoma tissue by Immunohistochemistry.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 showing positive staining in normal breast tissue.
  
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 showing positive staining in normal colon tissue.
  
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 showing positive staining in kidney carcinoma tissue.
  
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 showing negative staining in skeletal muscle tissue.
  
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 showing positive staining in urinary bladder transitional carcinoma tissue.
  
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Unpurified Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 showing negative staining in normal heart tissue.
  
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  OI-RD Scanning - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free (ab196357)

  Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 staining SLC2A1 in wild-type A549 cells, with negative expression in SLC2A1 knockout A549 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730 at 1 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab115730).