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st3gal1 (R&D Systems)

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R&D Systems st3gal1
St3gal1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/st3gal1/product/R&D Systems
Average 94 stars, based on 4 article reviews

st3gal1 - by Bioz Stars, 2026-03

94/100 stars

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(A) α-2,3 sialylated glycans generated by ST3GAL1 and ST3GAL2 and recognized by diCBM40 lectin. (B) Whisker plot illustrating significant upregulation of ST3GAL1 and S T3GAL2 mRNA expression in melanoma samples compared to nevi in multiple datasets: GSE3189 , GSE46517 , GSE12391 . Two-tailed unpaired t-test. (C) Representative images of IHC staining with ST3GAL1 and ST3GAL2 antibodies in 15 nevi and 50 melanoma samples show a perinuclear staining pattern (Fast red counterstaining). IHC score was calculated by combining the signal intensity and percentage of positive cells within the section. The histogram shows the distribution of ST3GAL1 and ST3GAL2 IHC scores in nevi and melanoma samples. Scale bar, 10 µm. (D) Representative images of diCBM40 lectin fluorescence microscopy of nevi and melanoma FFPE tissues (n = 17 for nevi and 68 for melanomas). diCBM40-Alexa 647 (magenta) and DAPI-stained sections of TMA. Scale bar, 100 µm. Dot plots represent the average fluorescence intensity of five fields per image, two-tailed unpaired t-test.

Journal: bioRxiv

Article Title: Integrated in vivo functional screens and multi-omics analyses identify α-2,3-sialylation as essential for melanoma maintenance

doi: 10.1101/2024.03.08.584072

Figure Lengend Snippet: (A) α-2,3 sialylated glycans generated by ST3GAL1 and ST3GAL2 and recognized by diCBM40 lectin. (B) Whisker plot illustrating significant upregulation of ST3GAL1 and S T3GAL2 mRNA expression in melanoma samples compared to nevi in multiple datasets: GSE3189 , GSE46517 , GSE12391 . Two-tailed unpaired t-test. (C) Representative images of IHC staining with ST3GAL1 and ST3GAL2 antibodies in 15 nevi and 50 melanoma samples show a perinuclear staining pattern (Fast red counterstaining). IHC score was calculated by combining the signal intensity and percentage of positive cells within the section. The histogram shows the distribution of ST3GAL1 and ST3GAL2 IHC scores in nevi and melanoma samples. Scale bar, 10 µm. (D) Representative images of diCBM40 lectin fluorescence microscopy of nevi and melanoma FFPE tissues (n = 17 for nevi and 68 for melanomas). diCBM40-Alexa 647 (magenta) and DAPI-stained sections of TMA. Scale bar, 100 µm. Dot plots represent the average fluorescence intensity of five fields per image, two-tailed unpaired t-test.

Article Snippet: Unconjugated anti-human ST3GAL1 and ST3GAL2, raised against human ST3GAL1 (Sigma HPA040466) and ST3GAL2 (Abcam ab96028), were used for IHC.

Techniques: Generated, Whisker Assay, Expressing, Two Tailed Test, Immunohistochemistry, Staining, Fluorescence, Microscopy

(A) Representative ST3GAL1 IHC images depict IHC scoring categories. ( B ) diCBM40 fluorescence microscopy of 5B1 cells with or without α-2,3-neuraminidase (NEB, P0743) treatment. 5B1 cells were incubated with his tagged diCBM40 after neuraminidase treatment and visualized by 6x-His Tag Monoclonal Antibody (HIS.H8), Alexa Fluor™ 647.

Journal: bioRxiv

Article Title: Integrated in vivo functional screens and multi-omics analyses identify α-2,3-sialylation as essential for melanoma maintenance

doi: 10.1101/2024.03.08.584072

Figure Lengend Snippet: (A) Representative ST3GAL1 IHC images depict IHC scoring categories. ( B ) diCBM40 fluorescence microscopy of 5B1 cells with or without α-2,3-neuraminidase (NEB, P0743) treatment. 5B1 cells were incubated with his tagged diCBM40 after neuraminidase treatment and visualized by 6x-His Tag Monoclonal Antibody (HIS.H8), Alexa Fluor™ 647.

Article Snippet: Unconjugated anti-human ST3GAL1 and ST3GAL2, raised against human ST3GAL1 (Sigma HPA040466) and ST3GAL2 (Abcam ab96028), were used for IHC.

Techniques: Fluorescence, Microscopy, Incubation

ST3GAL1 (A) and ST3GAL2 (B) protein levels in 5B1 and 12-273BM cells stably transduced with non-targeting scrambled control shRNA (shSCR), ST3GAL1 shRNAs (shA and shB) and ST3GAL2 shRNAs (shC and shD) were assessed by Western blotting. Western blot images are representative of three independent experiments. (C) Relative growth curves of 5B1 and 12-273BM cells stably transduced with non-targeting scrambled control shRNA (shSCR), ST3GAL1 shRNAs (shA and shB), and ST3GAL2 shRNAs (shC and shD). The data shown are representative of three independent experiments. Two-tailed unpaired t-tests and p-values are shown in the figures. (D) The percentage of melanoma cells positive for Annexin V only (early apoptosis) or PI only (necrosis), Experiment was performed in duplicates. (E) Representative Western blots for caspase 3 and PARP on lysates from 5B1 and 12-273BM cells with shRNA against ST3GAL1 and ST3GAL2.

Journal: bioRxiv

Article Title: Integrated in vivo functional screens and multi-omics analyses identify α-2,3-sialylation as essential for melanoma maintenance

doi: 10.1101/2024.03.08.584072

Figure Lengend Snippet: ST3GAL1 (A) and ST3GAL2 (B) protein levels in 5B1 and 12-273BM cells stably transduced with non-targeting scrambled control shRNA (shSCR), ST3GAL1 shRNAs (shA and shB) and ST3GAL2 shRNAs (shC and shD) were assessed by Western blotting. Western blot images are representative of three independent experiments. (C) Relative growth curves of 5B1 and 12-273BM cells stably transduced with non-targeting scrambled control shRNA (shSCR), ST3GAL1 shRNAs (shA and shB), and ST3GAL2 shRNAs (shC and shD). The data shown are representative of three independent experiments. Two-tailed unpaired t-tests and p-values are shown in the figures. (D) The percentage of melanoma cells positive for Annexin V only (early apoptosis) or PI only (necrosis), Experiment was performed in duplicates. (E) Representative Western blots for caspase 3 and PARP on lysates from 5B1 and 12-273BM cells with shRNA against ST3GAL1 and ST3GAL2.

Article Snippet: Unconjugated anti-human ST3GAL1 and ST3GAL2, raised against human ST3GAL1 (Sigma HPA040466) and ST3GAL2 (Abcam ab96028), were used for IHC.

Techniques: Stable Transfection, Transduction, shRNA, Western Blot, Two Tailed Test

( A and B ) ST3GAL1 and ST3GAL2 transcript expression in 5B1 and 12-273BM cells stably expressing shRNA targeting ST3GAL1 (shA or shB) or ST3GAL2 (shC or shD) or shSCR were assessed by real-time qPCR. qPCR graph shows average relative expression normalized to GAPDH , three replicates per condition, two-tailed unpaired t-tests. qPCR data are representative of three independent experiments. ( C ) diCBM40 fluorescence microscopy of 5B1 cells transduced by ST3GAL1 (shA or shB) or ST3GAL2 (shC or shD) or shSCR. 5B1 cells were visualized by incubating with diCBM40 lectin followed by 6x-His Tag Monoclonal Antibody (HIS.H8), Alexa Fluor™ 647. ( D ) Relative growth curves and ( E ) real-time qPCR of NHEM-A, A549, and HEK293T cells stably transduced with non-targeting scrambled control shRNA (shSCR), ST3GAL1 shRNAs (shA and shB) and ST3GAL2 shRNAs (shC and shD). ( F ) FACS analysis of cleaved caspase-3 on melanoma cells, 5B1 (left) and 12-273BM (right) transduced with two independent shST3GAL2 (shC and shD) treated with caspase inhibitor or vehicle.

Journal: bioRxiv

Article Title: Integrated in vivo functional screens and multi-omics analyses identify α-2,3-sialylation as essential for melanoma maintenance

doi: 10.1101/2024.03.08.584072

Figure Lengend Snippet: ( A and B ) ST3GAL1 and ST3GAL2 transcript expression in 5B1 and 12-273BM cells stably expressing shRNA targeting ST3GAL1 (shA or shB) or ST3GAL2 (shC or shD) or shSCR were assessed by real-time qPCR. qPCR graph shows average relative expression normalized to GAPDH , three replicates per condition, two-tailed unpaired t-tests. qPCR data are representative of three independent experiments. ( C ) diCBM40 fluorescence microscopy of 5B1 cells transduced by ST3GAL1 (shA or shB) or ST3GAL2 (shC or shD) or shSCR. 5B1 cells were visualized by incubating with diCBM40 lectin followed by 6x-His Tag Monoclonal Antibody (HIS.H8), Alexa Fluor™ 647. ( D ) Relative growth curves and ( E ) real-time qPCR of NHEM-A, A549, and HEK293T cells stably transduced with non-targeting scrambled control shRNA (shSCR), ST3GAL1 shRNAs (shA and shB) and ST3GAL2 shRNAs (shC and shD). ( F ) FACS analysis of cleaved caspase-3 on melanoma cells, 5B1 (left) and 12-273BM (right) transduced with two independent shST3GAL2 (shC and shD) treated with caspase inhibitor or vehicle.

Article Snippet: Unconjugated anti-human ST3GAL1 and ST3GAL2, raised against human ST3GAL1 (Sigma HPA040466) and ST3GAL2 (Abcam ab96028), were used for IHC.

Techniques: Expressing, Stable Transfection, shRNA, Two Tailed Test, Fluorescence, Microscopy, Transduction

$(A) Schematic illustration of the experimental approach showing affinity enrichment of α-2,3 sialylated proteins by MAA lectin affinity chromatography. (B) MAA affinity chromatography of whole-cell lysates of 5B1 cells transfected with NTC or ST3GAL1/ST3GAL2 shRNA followed by lectin blot with MAA lectin. An equal concentration of crude protein and an equal volume of MAA-enriched fractions was loaded for MAA lectin blot. (C) A number of proteins were identified by mass spectrometry analysis of the MAA-enriched fractions from 5B1, 12-273BN, and MeWo cell lines. ( D ) Gene ontology enrichment analysis (biological processes category) of α-2,3 sialylated proteins common to the three cell lines. Also, see (E) Western blot analysis with α-TFR1 or α-CD98hc antibodies of MAA-pulldown and corresponding input from lysates of 5B1 cells transfected with NTC or ST3GAL1/ST3GAL2 shRNA. (F) Western blot analysis of TFR1 and CD98hc in lysates from 5B1 cells transfected with NTC or ST3GAL1 or ST3GAL2 shRNAs. ( G ) Densitometric analysis of CD98hc on lysates from 5B1 cells transfected with NTC or ST3GAL1 or ST3GAL2 shRNAs. The graph is representative of three replicates. Experiments in B, E, and F were performed in triplicate, and representative images were shown.$

Journal: bioRxiv

Article Title: Integrated in vivo functional screens and multi-omics analyses identify α-2,3-sialylation as essential for melanoma maintenance

doi: 10.1101/2024.03.08.584072

Figure Lengend Snippet: (A) Schematic illustration of the experimental approach showing affinity enrichment of α-2,3 sialylated proteins by MAA lectin affinity chromatography. (B) MAA affinity chromatography of whole-cell lysates of 5B1 cells transfected with NTC or ST3GAL1/ST3GAL2 shRNA followed by lectin blot with MAA lectin. An equal concentration of crude protein and an equal volume of MAA-enriched fractions was loaded for MAA lectin blot. (C) A number of proteins were identified by mass spectrometry analysis of the MAA-enriched fractions from 5B1, 12-273BN, and MeWo cell lines. ( D ) Gene ontology enrichment analysis (biological processes category) of α-2,3 sialylated proteins common to the three cell lines. Also, see (E) Western blot analysis with α-TFR1 or α-CD98hc antibodies of MAA-pulldown and corresponding input from lysates of 5B1 cells transfected with NTC or ST3GAL1/ST3GAL2 shRNA. (F) Western blot analysis of TFR1 and CD98hc in lysates from 5B1 cells transfected with NTC or ST3GAL1 or ST3GAL2 shRNAs. ( G ) Densitometric analysis of CD98hc on lysates from 5B1 cells transfected with NTC or ST3GAL1 or ST3GAL2 shRNAs. The graph is representative of three replicates. Experiments in B, E, and F were performed in triplicate, and representative images were shown.

Article Snippet: Unconjugated anti-human ST3GAL1 and ST3GAL2, raised against human ST3GAL1 (Sigma HPA040466) and ST3GAL2 (Abcam ab96028), were used for IHC.

Techniques: Affinity Chromatography, Transfection, shRNA, Concentration Assay, Mass Spectrometry, Western Blot

( A ) Western blot for CD98hc on lysates from 5B1 cells transfected with NTC or CD98hc shRNAs. ( B ) Relative growth curves of 5B1 cells stably transduced with non-targeting control shRNA (shSCR) and CD98hc shRNAs (shA and shB). The data shown are representative of two independent experiments. Two-tailed unpaired t-tests and p-values are shown in the figures. ( C ) Western blot of CD98hc in 5B1 cells stably overexpressing CD98hc or control vector. ( D ) Cell proliferation assay on 5B1 melanoma cells stably overexpressing CD98hc or empty vector and transduced with non-targeting control shSCR or shST3GAL1. ( E ) Cell proliferation assay on 5B1 melanoma cells stably overexpressing CD98hc or empty vector and transduced with non-targeting control shSCR or shST3GAL2. ( F ) Western blot of CD98hc in 5B1 cells treated with or without 200 uM 3Fax-peracetylNeu5Ac. Densitometric analysis is shown below. ( G-H ) Western blot analysis of CD98hc protein in 5B1 cells. 3Fax-peracetylNeu5Ac treated 5B1 cells were further treated with 10 μM CHX at indicated intervals and analyzed by western blot analysis. The intensity of CD98hc protein was quantified using a densitometer. Western blot analysis of SLC3A2 protein in 5B1 cells silenced for ST3GAL1 ( I-J ) or ST3GAL2 ( K-L ) and treated with 10 μM cycloheximide (CHX) at indicated intervals and analyzed by western blot analysis. Paired t-test is shown. Western blot images are representative of 2 independent experiments.

Journal: bioRxiv

Article Title: Integrated in vivo functional screens and multi-omics analyses identify α-2,3-sialylation as essential for melanoma maintenance

doi: 10.1101/2024.03.08.584072

Figure Lengend Snippet: ( A ) Western blot for CD98hc on lysates from 5B1 cells transfected with NTC or CD98hc shRNAs. ( B ) Relative growth curves of 5B1 cells stably transduced with non-targeting control shRNA (shSCR) and CD98hc shRNAs (shA and shB). The data shown are representative of two independent experiments. Two-tailed unpaired t-tests and p-values are shown in the figures. ( C ) Western blot of CD98hc in 5B1 cells stably overexpressing CD98hc or control vector. ( D ) Cell proliferation assay on 5B1 melanoma cells stably overexpressing CD98hc or empty vector and transduced with non-targeting control shSCR or shST3GAL1. ( E ) Cell proliferation assay on 5B1 melanoma cells stably overexpressing CD98hc or empty vector and transduced with non-targeting control shSCR or shST3GAL2. ( F ) Western blot of CD98hc in 5B1 cells treated with or without 200 uM 3Fax-peracetylNeu5Ac. Densitometric analysis is shown below. ( G-H ) Western blot analysis of CD98hc protein in 5B1 cells. 3Fax-peracetylNeu5Ac treated 5B1 cells were further treated with 10 μM CHX at indicated intervals and analyzed by western blot analysis. The intensity of CD98hc protein was quantified using a densitometer. Western blot analysis of SLC3A2 protein in 5B1 cells silenced for ST3GAL1 ( I-J ) or ST3GAL2 ( K-L ) and treated with 10 μM cycloheximide (CHX) at indicated intervals and analyzed by western blot analysis. Paired t-test is shown. Western blot images are representative of 2 independent experiments.

Article Snippet: Unconjugated anti-human ST3GAL1 and ST3GAL2, raised against human ST3GAL1 (Sigma HPA040466) and ST3GAL2 (Abcam ab96028), were used for IHC.

Techniques: Western Blot, Transfection, Stable Transfection, Transduction, shRNA, Two Tailed Test, Plasmid Preparation, Proliferation Assay

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