Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
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.
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/
Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Lanes 1 - 2: Merged signal (red and green). Green - ab196357 observed at 54 kDa. Red - 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 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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed 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] (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 (ab261869) at 20 µg
Predicted band size: 54 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
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).
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/
Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Exposure time
Lane 1 to 2: 10 seconds
Lane 3 to 4: 30 seconds
We recommend not to boil the samples after lysis to get desired WB bands.
All lanes: Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) at 1/50000 dilution
Lane 1: HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates unboiled at 20 µg
Lane 2: HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates boiled at 20 µg
Lane 3: 3T3-L1 (Mouse embryonic fibroblast) whole cell lysates unboiled at 20 µg
Lane 4: 3T3-L1 (Mouse embryonic fibroblast) whole cell lysates boiled at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 54 kDa
Observed band size: 40-60 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Immunofluorescence staining of HepG2 cells with purified ab115730 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with 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 ab115730 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Alexa Fluor® 488 (ab195359) and Alexa Fluor® 647 (ab195020) conjugated versions are available for this clone.
Flow Cytometry (Intracellular) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified 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).
Alexa Fluorr®488 (ab195359) and Alexa Fluorr®647 (ab195020) conjugated versions are available for this clone.
