rabbit polyclonal anti glucose transporter 1  (Proteintech)

Journal: Nature Communications

Article Title: SPNS2 exports sphingosine-1-phosphate and imports glucose

doi: 10.1038/s41467-026-71659-7

Figure Lengend Snippet: a Immunofluorescence localization of SPNS2 (red) on plasma membrane of SVEC4-10. b Glucose increases S1P secretion by SVEC4-10 cells (n = 3, N = 3). c SVEC4-10 cells were treated with S1P lyase inhibitor A6770 (200 µM) or with SPNS2 inhibitor SLF1081851 (2 µM) and levels of S1P in cells (n = 8–10, N = 5) and medium (n = 9, N = 3) as well as glucose uptake were determined (n = 7, N = 3). d SPNS2 expression in two SPNS2 stably overexpressing SVEC4-10 cell lines generated by CRISPR activation plasmids (CTL1, SPNS2-OE1) or lentiviral activation particles (CTL2, SPNS2-OE2) and in two SPNS2 deleted SVEC4-10 cell lines (SPNS2-KO1 generated with double nickase plasmids, and SPNS2-KO2 via CRISPR/Cas9 knock-out and homology-directed repair plasmids) compared to their controls. e – h S1P levels in cells and medium, and glucose uptake were measured in SPNS2-OE1 ( e ), SPNS2-OE2 ( f ), SPNS2-KO1 ( g ), SPNS2-KO2 ( h ). (n = 6, N = 3). i , j SPNS2-OE1, SPNS2-KO1 cells and their controls were treated with S1P (500 nM), and phosphorylation of p42/44 ( i ) and glucose uptake ( j ) were determined (n = 6, N = 3). k , l Glucose uptake in SPNS2-OE1, SPNS2-KO1 and their control cells treated with insulin (100 nM) or GLUT1 inhibitor BAY-876 (10 nM) (n = 3, N = 3). Data are means ± s.d. b One-way analysis of variance test followed by Šídák’s multiple comparisons test. c – l two-tailed unpaired t-test. Source data are available for this figure in the Source Data file.

Article Snippet: To overexpress TurboGFP-tagged SPNS2 for ligand binding studies, SVEC4-10 cells were transfected with 2 μg of SPNS2 ( NM_001124758 ) Human Tagged ORF Clone in pCMV6-AC-GFP (#RG225940, Origene), 2 μg of Spns2 ( NM_153060 ) Mouse Tagged ORF Clone in pCMV6-AC-GFP (# MG218672 , Origene), or as controls with 2 μg of the pCMV6-AC-GFP vector (# PS100010 , Origene) or 2 μg of glucose transporter GLUT1 (SLC2A1) ( NM_006516 ), or Human Tagged ORF Clone in pCMV6-AC-GFP (#RG222696, Origene).

Techniques: Immunofluorescence, Clinical Proteomics, Membrane, Expressing, Stable Transfection, Generated, CRISPR, Activation Assay, Knock-Out, Phospho-proteomics, Control, Two Tailed Test

Journal: Nature Communications

Article Title: SPNS2 exports sphingosine-1-phosphate and imports glucose

doi: 10.1038/s41467-026-71659-7

Figure Lengend Snippet: a Representative image showing localization of SPNS2-TurboGFP and GLUT1-TurboGFP. b Temperature shift melting curves for purified hSPNS2-TurboGFP in the absence (blue) and presence of glucose (black). (n = 3, N = 3). Values are means ± s.d. c Apparent melting temperatures (T m ) for hSPNS2-TurboGFP, purified untagged hSPNS2 and purified mSPNS2-FLAG were calculated from the inflection points of the fitting curves. (n = 3, N = 3). Values are means ± s.e.m. Two-tailed unpaired t-test. d Schematic representation of the scintillation proximity assay (SPA). TurboGFP-tagged transport proteins are bound to scintillation beads. When radiolabeled glucose binds to these bead-bound transport proteins, emitted β-rays are close enough to stimulate the scintillation beads to emit light. Illustration created by Luciana Giono. e SPA signals of [ 3 H]glucose (0.8 μCi) binding to TurboGFP (vector), GLUT1-TurboGFP mSPNS2-TurboGFP, and hSPNS2-TurboGFP and its mutants E433A and T329A immobilized on the surfaces of polyvinyl toluene protein A-coated scintillation beads (500 μg per well) were measured by a scintillation counter. Values are means ± s.e.m. (n = 7; N = 2) and (n = 3; N = 3). One-way analysis of variance test followed by Dunnett’s multiple comparisons test. Source data are available for this figure in the Source Data file.

Article Snippet: To overexpress TurboGFP-tagged SPNS2 for ligand binding studies, SVEC4-10 cells were transfected with 2 μg of SPNS2 ( NM_001124758 ) Human Tagged ORF Clone in pCMV6-AC-GFP (#RG225940, Origene), 2 μg of Spns2 ( NM_153060 ) Mouse Tagged ORF Clone in pCMV6-AC-GFP (# MG218672 , Origene), or as controls with 2 μg of the pCMV6-AC-GFP vector (# PS100010 , Origene) or 2 μg of glucose transporter GLUT1 (SLC2A1) ( NM_006516 ), or Human Tagged ORF Clone in pCMV6-AC-GFP (#RG222696, Origene).

Techniques: Purification, Two Tailed Test, Scintillation Proximity Assay, Binding Assay, Plasmid Preparation

Journal: Nature Communications

Article Title: SPNS2 exports sphingosine-1-phosphate and imports glucose

doi: 10.1038/s41467-026-71659-7

Figure Lengend Snippet: a , b Identification of key SPNS2 residues involved in glucose transport. a Western blots and representative images of localization of hSPNS2-TurboGFP and its mutants. b Glucose uptake activities of SPNS2 variants with mutations in potential key residues involved in glucose or S1P engagement. hSPN S2 , GLUT1 vector, or the indicated mutants were overexpressed in SPNS2-KO1 cells lacking endogenous SPNS2. Glucose uptake was normalized to SPNS2 expression determined by GFP fluorescence ( n = 5, N = 3). Data are means ± s.e.m. One-way analysis of variance test followed by Dunnett’s multiple comparison test. c – f Direct glucose and S1P transport by SPNS2 proteoliposomes. c Illustration of cell-free preparation of SPNS2 proteoliposomes for functional transport analysis. d Comparable levels of hSPNS2 and its variants by western blots. e , f Proteoliposomes of WT hSPNS2 and variants were loaded without or with glucose ( e ) or S1P ( f ) as indicated and uptake of 1 μM NBD-S1P ( e ) or 1 µM NBD-glucose ( f ) determined. Arbitrary units (a.u.) (n = 3, N = 3). Nonspecific uptake was measured using protein-free liposomes (empty), vector containing proteoliposomes (vector), and T1R1 containing proteoliposomes (control). (n = 3, N = 3). One-way analysis of variance test followed by Dunnett’s multiple comparisons test. g Illustration of SPNS2-mediated export of S1P out of cells while transporting glucose inward. Illustrations in panels c and g created by Luciana Giono. h – j SPNS2-mediated D-[3H]glucose uptake. h Time-dependent specific uptake of D-[ 3 H]glucose into hSPNS2-containing proteoliposomes that were loaded without or with S1P (n = 3–5, N = 3). Nonspecific uptake measured using protein-free liposomes (empty) was subtracted from the specific uptake. i Uptake of [ 3 H]glucose by hSPNS2 or empty liposomes at 40 sec (n = 4, N = 3). j Kinetics of D-glucose uptake by hSPNS2. Specific uptake measured at 40 sec was calculated by subtraction of nonspecific [ 3 H]glucose uptake by empty liposomes and fitted to a non-linear regression analysis using Michaelis–Menten enzyme kinetics plot with K M , V max , and k cat values calculated (n = 4, N = 4). Data are means ± s.e.m. of independent experiments. Source data are available for this figure in the Source Data file.

Article Snippet: To overexpress TurboGFP-tagged SPNS2 for ligand binding studies, SVEC4-10 cells were transfected with 2 μg of SPNS2 ( NM_001124758 ) Human Tagged ORF Clone in pCMV6-AC-GFP (#RG225940, Origene), 2 μg of Spns2 ( NM_153060 ) Mouse Tagged ORF Clone in pCMV6-AC-GFP (# MG218672 , Origene), or as controls with 2 μg of the pCMV6-AC-GFP vector (# PS100010 , Origene) or 2 μg of glucose transporter GLUT1 (SLC2A1) ( NM_006516 ), or Human Tagged ORF Clone in pCMV6-AC-GFP (#RG222696, Origene).

Techniques: Western Blot, Plasmid Preparation, Expressing, Fluorescence, Comparison, Functional Assay, Liposomes, Control

Journal: BMC Cancer

Article Title: Glucose-transporter 1 (GLUT1) as a prognostic biomarker: evidence from 14,966 human tumors across 134 cancer types

doi: 10.1186/s12885-025-15527-5

Figure Lengend Snippet: Comparison of GLUT1 immunostaining results with previous GLUT1 studies. An „X“ indicates the fraction of GLUT1 positive cancer cells in the present study, dots indicate the reported frequencies from the literature for comparison: red dots mark studies with ≤10 analyzed tumors, yellow dots mark studies with ≥11 ≤ 25 analyzed tumors and green dots mark studies with > 25 analyzed tumors. All studies are listed in supplementary table 1. Locations of specific tumors types include seminoma of the testis, soft tissue rhabdoid tu-mors, teratomas of the testis, and typical as well as typical and atypical neuroendocrine of the lungs

Article Snippet: For the purpose of antibody validation, the normal tissue TMA was also analyzed by the rabbit polyclonal GLUT1 antibody 355 A-15 (Cell Marque, Rocklin, CA, pH 6.0, 1:200) on a DAKO autostainer Link48 according to a protocol suggested by Agilent DAKO.

Techniques: Comparison, Immunostaining

Journal: BMC Cancer

Article Title: Glucose-transporter 1 (GLUT1) as a prognostic biomarker: evidence from 14,966 human tumors across 134 cancer types

doi: 10.1186/s12885-025-15527-5

Figure Lengend Snippet: GLUT1 immunostaining in normal tissue. GLUT1 staining was typically membranous but also cytoplasmic. The panels show strong GLUT1 staining in amnion and chorion cells of the placenta ( A ), predominantly membranous staining in cyto- and syncytiotrophoblasts of a mature placenta ( B ), lack of GLUT1 staining of cerebral cells with a strong staining of small vessel cells in the grey cerebrum ( C ), moderate GLUT1 staining of suprabasal cell layers of the anal skin ( D ), focal weak GLUT1 staining in the surface epithelium of the gallbladder ( E ), weak to moderate GLUT1 staining of few collecting ducts in the renal cortex ( F ), weak GLUT1 staining in basal cells of the prostate ( G ), and lack of GLUT1 staining in sinusoidal cells with strong staining of erythrocytes in the liver ( H )

Article Snippet: For the purpose of antibody validation, the normal tissue TMA was also analyzed by the rabbit polyclonal GLUT1 antibody 355 A-15 (Cell Marque, Rocklin, CA, pH 6.0, 1:200) on a DAKO autostainer Link48 according to a protocol suggested by Agilent DAKO.

Techniques: Immunostaining, Staining

Journal: BMC Cancer

Article Title: Glucose-transporter 1 (GLUT1) as a prognostic biomarker: evidence from 14,966 human tumors across 134 cancer types

doi: 10.1186/s12885-025-15527-5

Figure Lengend Snippet: GLUT1 immunostaining in cancer. Strong GLUT1 immunostaining in clear cell renal cell carcinoma ( A ), urothelial carcinoma ( B ), colorectal carcinoma ( C ), serous high-grade ovarian carcinoma ( D ), squamous cell carcinoma of the vulva ( E ), endometrioid endometrial carcinoma ( F ), enbryonel carcinoma of the testis ( G ), and absence of GLUT1 immunostaining in hepatocellular carcinoma ( H )

Article Snippet: For the purpose of antibody validation, the normal tissue TMA was also analyzed by the rabbit polyclonal GLUT1 antibody 355 A-15 (Cell Marque, Rocklin, CA, pH 6.0, 1:200) on a DAKO autostainer Link48 according to a protocol suggested by Agilent DAKO.

Techniques: Immunostaining

Journal: BMC Cancer

Article Title: Glucose-transporter 1 (GLUT1) as a prognostic biomarker: evidence from 14,966 human tumors across 134 cancer types

doi: 10.1186/s12885-025-15527-5

Figure Lengend Snippet: Ranking order of GLUT1 immunostaining in tumors. Both the percentage of positive cases (blue dots) and the percentage of strongly positive cases (orange dots) are shown

Article Snippet: For the purpose of antibody validation, the normal tissue TMA was also analyzed by the rabbit polyclonal GLUT1 antibody 355 A-15 (Cell Marque, Rocklin, CA, pH 6.0, 1:200) on a DAKO autostainer Link48 according to a protocol suggested by Agilent DAKO.

Techniques: Immunostaining

Journal: BMC Cancer

Article Title: Glucose-transporter 1 (GLUT1) as a prognostic biomarker: evidence from 14,966 human tumors across 134 cancer types

doi: 10.1186/s12885-025-15527-5

Figure Lengend Snippet: GLUT1 immunostaining and prognosis in ( A - D ) clear cell renal cell cancer and in ( E - H ) papillary renal cell cancer. A , B ), E ), and F ) show overall survival, C ), D ), G ) and H ) show recurrence free survival. For B ), D ), F and H ), tumors were grouped in GLUT1 “low”, including negative, weak and moderate staining, and GLUT1 “high” (strong) staining

Article Snippet: For the purpose of antibody validation, the normal tissue TMA was also analyzed by the rabbit polyclonal GLUT1 antibody 355 A-15 (Cell Marque, Rocklin, CA, pH 6.0, 1:200) on a DAKO autostainer Link48 according to a protocol suggested by Agilent DAKO.

Techniques: Immunostaining, Staining

Journal: Histochemistry and Cell Biology

Article Title: Immunohistochemical analysis to detect a molecular signature in intervertebral disc degeneration

doi: 10.1007/s00418-025-02434-w

Figure Lengend Snippet: Detection of FOXO3a ( a ), SOD2 ( b ), HIF1α ( c ), GLUT1 ( d ), and Bry ( e ) expression by immunohistochemistry. Immunohistochemical analysis was performed on IVD tissues with different Pfirrmann grades of degeneration. Protein levels were quantified by densitometric analysis of immunostaining using ImageJ software and expressed as the percentage of positive cells per area (five sections per sample; Pfirrmann I–II group, n = 6; Pfirrmann III group, n = 40 and Pfirrmann IV–V group, n = 9). Results are reported as a whisker box plot representing the min to max (the line indicates median). * p < 0.01 (Pfirrmann III group vs. Pfirrmann I–II group); ^ p < 0.01 (Pfirrmann IV–V group vs. Pfirrmann I–II group); § p < 0.01 (Pfirrmann III group vs. Pfirrmann IV–V group). Scale bars 20 μm

Article Snippet: For immunohistochemical evaluation, sections were incubated overnight (4 °C) with a primary antibody against FOXO3a (#ab70315, rabbit anti-human, 1:100 dilution; Abcam, Cambridge, UK), SOD2 (#sc-133134, mouse anti-human, 1:100 dilution; Santa Cruz biotech., Dallas, USA), HIF1α (clone H1alpha67, mouse anti-human, 1:200 dilution; Novusbio, Centennial, USA), GLUT1 (#TA301678, mouse anti-human, 1:200 dilution; Origene, Rockville, USA), and BRY (#ab209665, rabbit anti-human, 1:500 dilution; Abcam), followed by treatment with Vectastain ABC solution (#MP-7500; Vectorlabs, Burlingame, USA) for 30 min. To achieve specificity, the working concentrations of primary antibodies were selected by following the dilution guidelines and technical specifications provided by the antibody’s manufacturer.

Techniques: Expressing, Immunohistochemistry, Immunohistochemical staining, Immunostaining, Software, Whisker Assay

Journal: Histochemistry and Cell Biology

Article Title: Immunohistochemical analysis to detect a molecular signature in intervertebral disc degeneration

doi: 10.1007/s00418-025-02434-w

Figure Lengend Snippet: Analysis of FOXO3a, SOD2, HIF1α, GLUT1, and Bry expression in IVD tissues with Pfirrmann grade III. Protein levels were assessed by immunohistochemistry and quantified by densitometric analysis of immunostaining using ImageJ software. Results were expressed as the percentage of positive cells per area, based on five representative sections per sample ( n = 40, Pfirrmann grade III). Data are presented as box-and-whisker plots, displaying min to max (the line indicates the median). Comparisons were made on the basis of the following parameters: a sex (female, male), age (18–40, 41–60, > 60 years), smoking status (non-smoker [NS], former smoker [FS], current smoker [CS]), and body mass index (BMI) (normal weight [NW], overweight [OW], obese [OB]); b anatomical site of surgery (L2–L3, L3–L4, L4–L5, L5–S1), duration of symptoms before surgery (< 6 months, ≥ 6 months), and area of infiltration (moderate, abundant)

Article Snippet: For immunohistochemical evaluation, sections were incubated overnight (4 °C) with a primary antibody against FOXO3a (#ab70315, rabbit anti-human, 1:100 dilution; Abcam, Cambridge, UK), SOD2 (#sc-133134, mouse anti-human, 1:100 dilution; Santa Cruz biotech., Dallas, USA), HIF1α (clone H1alpha67, mouse anti-human, 1:200 dilution; Novusbio, Centennial, USA), GLUT1 (#TA301678, mouse anti-human, 1:200 dilution; Origene, Rockville, USA), and BRY (#ab209665, rabbit anti-human, 1:500 dilution; Abcam), followed by treatment with Vectastain ABC solution (#MP-7500; Vectorlabs, Burlingame, USA) for 30 min. To achieve specificity, the working concentrations of primary antibodies were selected by following the dilution guidelines and technical specifications provided by the antibody’s manufacturer.

Techniques: Expressing, Immunohistochemistry, Immunostaining, Software, Whisker Assay

Journal: Histochemistry and Cell Biology

Article Title: Immunohistochemical analysis to detect a molecular signature in intervertebral disc degeneration

doi: 10.1007/s00418-025-02434-w

Figure Lengend Snippet: Correlation between FOXO3a, SOD2, HIF1α, GLUT1, and Bry expression levels and the onset of chronic pain or relapse. Results were expressed as percentage of positive cells per area, based on five representative sections per sample ( n = 24, Pfirrmann grade III). Data are presented as box-and-whisker plots, displaying min to max (the line indicates the median). Patients without chronic pain after surgery (green), with joint inflammatory symptoms after several months (blue), and with short-term relapses (red)

Article Snippet: For immunohistochemical evaluation, sections were incubated overnight (4 °C) with a primary antibody against FOXO3a (#ab70315, rabbit anti-human, 1:100 dilution; Abcam, Cambridge, UK), SOD2 (#sc-133134, mouse anti-human, 1:100 dilution; Santa Cruz biotech., Dallas, USA), HIF1α (clone H1alpha67, mouse anti-human, 1:200 dilution; Novusbio, Centennial, USA), GLUT1 (#TA301678, mouse anti-human, 1:200 dilution; Origene, Rockville, USA), and BRY (#ab209665, rabbit anti-human, 1:500 dilution; Abcam), followed by treatment with Vectastain ABC solution (#MP-7500; Vectorlabs, Burlingame, USA) for 30 min. To achieve specificity, the working concentrations of primary antibodies were selected by following the dilution guidelines and technical specifications provided by the antibody’s manufacturer.

Techniques: Expressing, Whisker Assay