Immunocytochemistry:Article Title: The calcium binding protein S100β marks hedgehog-responsive resident vascular stem cells within vascular lesions
Article Snippet: .. Rat anti-mouse Sca1/Ly6A/E antibody [D7] (FITC) antibody Abcam (ab25031) 1/200 Rabbit anti-mouse/rat alpha smooth muscle cell alpha-actin antibody Abcam (ab5694) 1/200 Anti-Patched / PTCH1 antibody, Mouse monoclonal Abcam (Ab55629) 1/100 Anti-actin, α-Smooth Muscle antibody, Mouse monoclonal Sigma (A5228) 1/200 Anti-S100-β (CT) Antibody, clone EP1576Y, rabbit monoclonal Millipore (04-1054) 1/100 Anti-Gli2 antibody, rabbit polyclonal Novus Biologicals (NBP2-23602SS) 1/50 anti–alpha-smooth muscle actin (α-SMA) antibody (Abcam ab7817), 1/200 anti-eNOS antibody (Abcam ab76198) 1/200 anti-CD31 (Abcam ab24590) 1/100 Chicken anti-GFP antibody Abcam (ab13970) 1/500 Rabbit Anti-RFP/dT antibody Abcam (ab62341) 1/500 Goat anti-rabbit IgG secondary Alexa Fluor 647® conjugate Invitrogen (Cat # S32357) 1/1000 Antibodies and their corresponding dilutions used in immunocytochemistry Mouse anti-mouse/rat nestin [Rat-401] Abcam (ab11306) 1/200 Rabbit anti-mouse Calponin [EP798Y] Abcam (ab46794) 1/200 Goat anti-mouse/rat/human smooth muscle Myosin heavy chain Santa Cruz (sc-79079) 1/200 Mouse anti-human/rat SOX10 R&D System (MAB2864) 1/100 3 nature research | reporting sum m ary April 2020 Mouse anti-human SOX17 R&D System (MAB1924) 1/100 Rabbit Anti-S100β Merck Millipore (ABN59) 1/100 Rabbit Anti-mouse/rat S100β [EP1576Y] Abcam (ab52642) 1/100 Rabbit anti-mouse/rat Sca1 Millipore (AB4336) 1/100 Rabbit polyclonal anti-Oct4, Abcam (ab18976)1/100 Recombinant Anti-S100 beta antibody Abcam (ab52642) [EP1576Y] Recombinant Anti-Calponin 1 antibody [EP798Y] (ab46794) 1/100 Alexa Fluor 488 Goat anti-mouse IgG Invitrogen (A-11001) 1/1000 Alexa Fluor 488 Goat anti-rabbit IgG Invitrogen (A-11008) 1/1000 Alexa Fluor 488 Donkey anti-goat IgG Invitrogen (A-11055) 1/1000 Antibodies used in Chromatin Immunoprecipitation (ChIP) Rabbit anti-mouse Tri-Methyl-Histone H3 (Lys27) [C36B11] Cell Signalling Technology (9733S) Rabbit anti-mouse Di-Methyl-Histone H3 (Lys4) [C64G9] Cell Signalling Technology (9725S) Normal Rabbit IgG (ChIP graded) Cell Signalling Technology (2729) Antibodies and their corresponding dilutions used in Flow Cytometry. .. Rat anti-mouse Sca1 (Ly-6A/E) [E13-161.7] STEMCELL Technology (60032) 1/100 Rat anti-mouse IgG2a, kappa Isotype [RTK2758] STEMCELL Technology (60076) 1/100 Rabbit Anti-mouse/rat S100β [EP1576Y] Abcam (ab52642) 1/100 Normal Rabbit IgG (F graded) Cell Signalling Technology (2729) 1/100 Alexa Fluor 647 Goat anti-mouse (H+L) Life Technologies (A- A-21235) 1/100 Validation For validation, we introduced background blocking controls as well as specificity controls that included secondary antibodies alone.
Article Title: The calcium binding protein S100β marks hedgehog-responsive resident vascular stem cells within vascular lesions
Article Snippet: .. Antibody/Product Name Supplier/Product Number Dilutions Rat anti-mouse Sca1/Ly6A/E antibody [D7] (FITC) antibody Abcam (ab25031) 1/200 Rabbit anti-mouse/rat alpha smooth muscle cell ⍺-actin antibody Abcam (ab5694) 1/200 Anti-Patched / PTCH1 antibody, Mouse monoclonal Abcam (Ab55629) 1/100 Anti-Actin, α-Smooth Muscle antibody, Mouse monoclonal Sigma (A5228) 1/200 Anti-S100-β (CT) Antibody, clone EP1576Y, rabbit monoclonal Millipore (04-1054) 1/100 Anti-Gli2 antibody, rabbit polyclonal Novus Biologicals (NBP2-23602SS) 1/50 Chicken anti-GFP antibody Abcam (ab13970) 1/1000 Rabbit Anti-RFP/dT antibody Abcam (ab62341) 1/1000 Supplementary Table II Antibodies and their corresponding dilutions used in immunocytochemistry/western blot. .. Antibody/Product Name Supplier/Product Number Dilutions Mouse anti-mouse/rat nestin [Rat-401] Abcam (ab11306) 1/200 Rabbit anti-mouse Calponin [EP798Y] Abcam (ab46794) 1/200 Goat anti-mouse/rat/human smooth muscle Myosin heavy chain Santa Cruz (sc-79079) 1/200 Mouse anti-human/rat SOX10 R&D System (MAB2864) 1/100 Mouse anti-human SOX17 R&D System (MAB1924) 1/100 Rabbit Anti-S100β Merck Millipore (ABN59) 1/100 Rabbit Anti-mouse/rat S100β [EP1576Y] Abcam (ab52642) 1/100 Rabbit anti-mouse/rat Sca1 Millipore (AB4336) 1/100 Alexa Fluorâ 488 Goat anti-mouse IgG Invitrogen (A-11001) 1/1000 Alexa Fluorâ 488 Goat anti-rabbit IgG Invitrogen (A-11008) 1/1000 Alexa Fluorâ 488 Donkey anti-goat IgG Invitrogen (A-11055) 1/1000 Supplementary Table III Antibodies used in Chromatin Immunoprecipitation (ChIP) Antibody/Product Name Supplier/Product Number Rabbit anti-mouse Tri-Methyl-Histone H3 (Lys27) [C36B11] Cell Signalling Technology (9733S) Rabbit anti-mouse Di-Methyl-Histone H3 (Lys4) [C64G9] Cell Signalling Technology (9725S) Normal Rabbit IgG (ChIP graded) Cell Signalling Technology (2729) Supplementary Table IV Customised primers used in this study from Integrity DNA Technology (IDT).
Recombinant:Article Title: The calcium binding protein S100β marks hedgehog-responsive resident vascular stem cells within vascular lesions
Article Snippet: .. Rat anti-mouse Sca1/Ly6A/E antibody [D7] (FITC) antibody Abcam (ab25031) 1/200 Rabbit anti-mouse/rat alpha smooth muscle cell alpha-actin antibody Abcam (ab5694) 1/200 Anti-Patched / PTCH1 antibody, Mouse monoclonal Abcam (Ab55629) 1/100 Anti-actin, α-Smooth Muscle antibody, Mouse monoclonal Sigma (A5228) 1/200 Anti-S100-β (CT) Antibody, clone EP1576Y, rabbit monoclonal Millipore (04-1054) 1/100 Anti-Gli2 antibody, rabbit polyclonal Novus Biologicals (NBP2-23602SS) 1/50 anti–alpha-smooth muscle actin (α-SMA) antibody (Abcam ab7817), 1/200 anti-eNOS antibody (Abcam ab76198) 1/200 anti-CD31 (Abcam ab24590) 1/100 Chicken anti-GFP antibody Abcam (ab13970) 1/500 Rabbit Anti-RFP/dT antibody Abcam (ab62341) 1/500 Goat anti-rabbit IgG secondary Alexa Fluor 647® conjugate Invitrogen (Cat # S32357) 1/1000 Antibodies and their corresponding dilutions used in immunocytochemistry Mouse anti-mouse/rat nestin [Rat-401] Abcam (ab11306) 1/200 Rabbit anti-mouse Calponin [EP798Y] Abcam (ab46794) 1/200 Goat anti-mouse/rat/human smooth muscle Myosin heavy chain Santa Cruz (sc-79079) 1/200 Mouse anti-human/rat SOX10 R&D System (MAB2864) 1/100 3 nature research | reporting sum m ary April 2020 Mouse anti-human SOX17 R&D System (MAB1924) 1/100 Rabbit Anti-S100β Merck Millipore (ABN59) 1/100 Rabbit Anti-mouse/rat S100β [EP1576Y] Abcam (ab52642) 1/100 Rabbit anti-mouse/rat Sca1 Millipore (AB4336) 1/100 Rabbit polyclonal anti-Oct4, Abcam (ab18976)1/100 Recombinant Anti-S100 beta antibody Abcam (ab52642) [EP1576Y] Recombinant Anti-Calponin 1 antibody [EP798Y] (ab46794) 1/100 Alexa Fluor 488 Goat anti-mouse IgG Invitrogen (A-11001) 1/1000 Alexa Fluor 488 Goat anti-rabbit IgG Invitrogen (A-11008) 1/1000 Alexa Fluor 488 Donkey anti-goat IgG Invitrogen (A-11055) 1/1000 Antibodies used in Chromatin Immunoprecipitation (ChIP) Rabbit anti-mouse Tri-Methyl-Histone H3 (Lys27) [C36B11] Cell Signalling Technology (9733S) Rabbit anti-mouse Di-Methyl-Histone H3 (Lys4) [C64G9] Cell Signalling Technology (9725S) Normal Rabbit IgG (ChIP graded) Cell Signalling Technology (2729) Antibodies and their corresponding dilutions used in Flow Cytometry. .. Rat anti-mouse Sca1 (Ly-6A/E) [E13-161.7] STEMCELL Technology (60032) 1/100 Rat anti-mouse IgG2a, kappa Isotype [RTK2758] STEMCELL Technology (60076) 1/100 Rabbit Anti-mouse/rat S100β [EP1576Y] Abcam (ab52642) 1/100 Normal Rabbit IgG (F graded) Cell Signalling Technology (2729) 1/100 Alexa Fluor 647 Goat anti-mouse (H+L) Life Technologies (A- A-21235) 1/100 Validation For validation, we introduced background blocking controls as well as specificity controls that included secondary antibodies alone.
Article Title: Ergothioneine improves healthspan of aged animals by enhancing cGPDH activity through CSE-dependent persulfidation.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-4-HNE, rabbit polyclonal Invitrogen Cat# BS-6313R; RRID: AB_2827741 anti-SIRT1, rabbit polyclonal Proteintech Cat# 13161-1-AP; RRID: AB_10646436 anti-TOM20, rabbit polyclonal Proteintech Cat# 11802-1-AP; RRID: AB_2207530 anti-GPD1, rabbit polyclonal Proteintech Cat# 13451-1-AP; RRID: AB_10646471 anti-PGC1 alpha, rabbit polyclonal Novus Biologicals Cat# NBP1-04676; RRID: AB_1522118 anti-VEGFR2, mouse monoclonal Santa Cruz Biotechnology Cat# sc-6251; RRID: AB_628431 anti-GAPDH, mouse monoclonal Santa Cruz Biotechnology Cat# sc-47724; RRID: AB_627678 anti-PAX-7, mouse monoclonal Santa Cruz Biotechnology Cat# sc-81648; RRID: AB_2159836 WGA-AF488 Invitrogen Cat# W11261 Anti-Ac-Lysine, mouse monoclonal Santa Cruz Biotechnology Cat# sc-32268; RRID: AB_627898 anti- b-tubulin Sigma Aldrich Cat# T0198; RRID: AB_477556 anti-CSE Proteintech Cat# 12217-1-AP; RRID: AB_2087497 anti-Vinculin Invitrogen Cat# MA5-11690; RRID: AB_10976821 anti-S6 Cell Signaling Technology Cat# 2217; RRID: AB_331355 anti-pS6 Cell Signaling Technology Cat# 2211; RRID: AB_331679 anti-mouse-AF488, goat polyclonal Abcam Cat# ab150113; RRID: AB_2576208 anti-mouse-AF680, goat polyclonal Abcam Cat# ab175775 anti-rabbit-AF680, donkey polyclonal Abcam Cat# ab175772 anti-rabbit-CY5, goat polyclonal Invitrogen Cat# A10523; RRID: AB_2534032 anti-rabbit-HRP Abcam Cat# ab205718; RRID: AB_2819160 anti-mouse-HRP Cell Signaling Technology Cat# 7076S;RRID: AB_330924 Bacterial and virus strains OP50-1 Caenorhabditis Genetics Center N/A Chemicals, peptides, and recombinant proteins Chelex-100 Sigma Aldrich Cat# C7901 HEPES VWR Cat# 441485H 1M HEPES solution Sigma Aldrich Cat# H0887 EDTA Sigma Aldrich Cat# E5134 Neocuproine Sigma Aldrich Cat# N1501 IGEPAL Thermofisher Cat# J61055 SDS Carl-Roth Cat# 2326.2 TRIS Carl-Roth Cat# 5429.3 Sodium deoxycholate Sigma Aldrich Cat# D6750 Glycine Carl-Roth Cat# 3187.5 Ponceau S Sigma Aldrich Cat# P3504 Tween Sigma Aldrich Cat# P9416 Triton X-100 Sigma Aldrich Cat# T8787 Protease inhibitor cocktail Sigma Aldrich Cat# P8340 4-Chloro-7-nitrobenzofurazan (NBF-Cl) Sigma Aldrich Cat# 163260 1,4-dithiothreitol (DTT) ThermoFisher Cat# R0861 Iodoacetamide Sigma-Aldrich Cat# I1149 Ammonium bicarbonate Sigma Aldrich Cat# 09830 Calcium chloride Euromedex Cat# T885-A Guanidine hydrochloride Sigma Aldrich Cat# G3272 (Continued on next page) e1 Cell Metabolism 37, 1–15.e1–e14, February 4, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Triethylammonium bicarbonate buffer Sigma Aldrich Cat# 18597 Hydroxylamine solution Sigma Aldrich Cat# 467804 Trypsin Promega Cat# V5117 TMT10plex Thermo Scientific Cat# 90110 DCP-Bio1 Merck Cat# NS1226 Copper (II)-TBTA complex Lumiprobe Cat# 21050 L-ascorbic acid Sigma-Aldrich Cat# A92902 Formic acid Suprapur Supelco Cat# 111670 TCEP Sigma-Aldrich Cat# 646547 AQC Synchem Cat# s041 AccQ-Tag Ultra Borate Buffer Waters Cat# 186009283 Cyanine5 alkyne Lumirpobe Cat# A30B0 Laemmli (4X) buffer BioRad Cat# 1610747 b-mercaptoethanol Sigma Aldrich Cat# M3148 Western Blotting Luminol Reagent Santa Cruz Biotechnology Cat# sc-2048 Ponceau S Sigma Aldrich Cat# P3504 Hydrogen Peroxide Solution Sigma Aldrich Cat# 216763 Erastin Sigma Aldrich Cat# E7781 1S,3R-RSL 3 Sigma Aldrich Cat# SML2234 Thiazolyl Blue Tetrazolium Bromide Sigma Aldrich Cat# M5655 Methyl viologen dichloride hydrate Sigma Aldrich Cat# 856177 Sodium (meta)arsenite Santa Cruz Biotechnology Cat# sc-250986 Nile Red Sigma Aldrich Cat# 72485 Oil Red O Sigma Aldrich Cat# O0625 Bovine Serum Albumin Sigma Aldrich Cat# A7906 L-(+)-Ergothioneine Thermo Scientific Cat# J67861.03 L-Hercynine Sigma Aldrich Cat# SMB00978 DMEM Gibco Cat# 21969-035 DMEM, high glucose, no glutamine, no methionine, no cystine Gibco Cat# 21969-035 DMEM/F-12 Gibco Cat# 21041025 DMEM, no phenol-red Gibco Cat# 31053028 Endothelial Cell Growth Medium 2 (ECGM2) PromoCell Cat# C-22011 Fetal bovine serum Pan Biotech Cat# P30-3031 L-glutamine Pan Biotech Cat# P64-80100 Penicillin-streptomycin Pan Biotech Cat# P66-07100 Polybrene Santa Cruz Biotechnology Cat# 28728-55-4 Lipofectamine RNAiMAX Invitrogen Cat# 13778075 DPBS Sigma Aldrich Cat# D8537 Hoechst 33342 Sigma Aldrich Cat# B2261 L-Cystine Sigma Aldrich Cat# 30200 Pierce High Capacity NeutrAvidin Agarose beads Thermofisher Cat# 20357 Pierce NeutrAvidin Agarose beads Thermofisher Cat# 29200 DTAB Thermo Scientific Cat# A10761.09 Sodium disulfide Dojindo Molecular Technologies Cat# SB-02 Dihydroxyacetone phosphate hemimagnesium salt hydrate Sigma Aldrich Cat# 51269 (Continued on next page) Cell Metabolism 37, 1–15.e1–e14, February 4, 2025 e2
Chromatin Immunoprecipitation:Article Title: The calcium binding protein S100β marks hedgehog-responsive resident vascular stem cells within vascular lesions
Article Snippet: .. Rat anti-mouse Sca1/Ly6A/E antibody [D7] (FITC) antibody Abcam (ab25031) 1/200 Rabbit anti-mouse/rat alpha smooth muscle cell alpha-actin antibody Abcam (ab5694) 1/200 Anti-Patched / PTCH1 antibody, Mouse monoclonal Abcam (Ab55629) 1/100 Anti-actin, α-Smooth Muscle antibody, Mouse monoclonal Sigma (A5228) 1/200 Anti-S100-β (CT) Antibody, clone EP1576Y, rabbit monoclonal Millipore (04-1054) 1/100 Anti-Gli2 antibody, rabbit polyclonal Novus Biologicals (NBP2-23602SS) 1/50 anti–alpha-smooth muscle actin (α-SMA) antibody (Abcam ab7817), 1/200 anti-eNOS antibody (Abcam ab76198) 1/200 anti-CD31 (Abcam ab24590) 1/100 Chicken anti-GFP antibody Abcam (ab13970) 1/500 Rabbit Anti-RFP/dT antibody Abcam (ab62341) 1/500 Goat anti-rabbit IgG secondary Alexa Fluor 647® conjugate Invitrogen (Cat # S32357) 1/1000 Antibodies and their corresponding dilutions used in immunocytochemistry Mouse anti-mouse/rat nestin [Rat-401] Abcam (ab11306) 1/200 Rabbit anti-mouse Calponin [EP798Y] Abcam (ab46794) 1/200 Goat anti-mouse/rat/human smooth muscle Myosin heavy chain Santa Cruz (sc-79079) 1/200 Mouse anti-human/rat SOX10 R&D System (MAB2864) 1/100 3 nature research | reporting sum m ary April 2020 Mouse anti-human SOX17 R&D System (MAB1924) 1/100 Rabbit Anti-S100β Merck Millipore (ABN59) 1/100 Rabbit Anti-mouse/rat S100β [EP1576Y] Abcam (ab52642) 1/100 Rabbit anti-mouse/rat Sca1 Millipore (AB4336) 1/100 Rabbit polyclonal anti-Oct4, Abcam (ab18976)1/100 Recombinant Anti-S100 beta antibody Abcam (ab52642) [EP1576Y] Recombinant Anti-Calponin 1 antibody [EP798Y] (ab46794) 1/100 Alexa Fluor 488 Goat anti-mouse IgG Invitrogen (A-11001) 1/1000 Alexa Fluor 488 Goat anti-rabbit IgG Invitrogen (A-11008) 1/1000 Alexa Fluor 488 Donkey anti-goat IgG Invitrogen (A-11055) 1/1000 Antibodies used in Chromatin Immunoprecipitation (ChIP) Rabbit anti-mouse Tri-Methyl-Histone H3 (Lys27) [C36B11] Cell Signalling Technology (9733S) Rabbit anti-mouse Di-Methyl-Histone H3 (Lys4) [C64G9] Cell Signalling Technology (9725S) Normal Rabbit IgG (ChIP graded) Cell Signalling Technology (2729) Antibodies and their corresponding dilutions used in Flow Cytometry. .. Rat anti-mouse Sca1 (Ly-6A/E) [E13-161.7] STEMCELL Technology (60032) 1/100 Rat anti-mouse IgG2a, kappa Isotype [RTK2758] STEMCELL Technology (60076) 1/100 Rabbit Anti-mouse/rat S100β [EP1576Y] Abcam (ab52642) 1/100 Normal Rabbit IgG (F graded) Cell Signalling Technology (2729) 1/100 Alexa Fluor 647 Goat anti-mouse (H+L) Life Technologies (A- A-21235) 1/100 Validation For validation, we introduced background blocking controls as well as specificity controls that included secondary antibodies alone.
Flow Cytometry:Article Title: The calcium binding protein S100β marks hedgehog-responsive resident vascular stem cells within vascular lesions
Article Snippet: .. Rat anti-mouse Sca1/Ly6A/E antibody [D7] (FITC) antibody Abcam (ab25031) 1/200 Rabbit anti-mouse/rat alpha smooth muscle cell alpha-actin antibody Abcam (ab5694) 1/200 Anti-Patched / PTCH1 antibody, Mouse monoclonal Abcam (Ab55629) 1/100 Anti-actin, α-Smooth Muscle antibody, Mouse monoclonal Sigma (A5228) 1/200 Anti-S100-β (CT) Antibody, clone EP1576Y, rabbit monoclonal Millipore (04-1054) 1/100 Anti-Gli2 antibody, rabbit polyclonal Novus Biologicals (NBP2-23602SS) 1/50 anti–alpha-smooth muscle actin (α-SMA) antibody (Abcam ab7817), 1/200 anti-eNOS antibody (Abcam ab76198) 1/200 anti-CD31 (Abcam ab24590) 1/100 Chicken anti-GFP antibody Abcam (ab13970) 1/500 Rabbit Anti-RFP/dT antibody Abcam (ab62341) 1/500 Goat anti-rabbit IgG secondary Alexa Fluor 647® conjugate Invitrogen (Cat # S32357) 1/1000 Antibodies and their corresponding dilutions used in immunocytochemistry Mouse anti-mouse/rat nestin [Rat-401] Abcam (ab11306) 1/200 Rabbit anti-mouse Calponin [EP798Y] Abcam (ab46794) 1/200 Goat anti-mouse/rat/human smooth muscle Myosin heavy chain Santa Cruz (sc-79079) 1/200 Mouse anti-human/rat SOX10 R&D System (MAB2864) 1/100 3 nature research | reporting sum m ary April 2020 Mouse anti-human SOX17 R&D System (MAB1924) 1/100 Rabbit Anti-S100β Merck Millipore (ABN59) 1/100 Rabbit Anti-mouse/rat S100β [EP1576Y] Abcam (ab52642) 1/100 Rabbit anti-mouse/rat Sca1 Millipore (AB4336) 1/100 Rabbit polyclonal anti-Oct4, Abcam (ab18976)1/100 Recombinant Anti-S100 beta antibody Abcam (ab52642) [EP1576Y] Recombinant Anti-Calponin 1 antibody [EP798Y] (ab46794) 1/100 Alexa Fluor 488 Goat anti-mouse IgG Invitrogen (A-11001) 1/1000 Alexa Fluor 488 Goat anti-rabbit IgG Invitrogen (A-11008) 1/1000 Alexa Fluor 488 Donkey anti-goat IgG Invitrogen (A-11055) 1/1000 Antibodies used in Chromatin Immunoprecipitation (ChIP) Rabbit anti-mouse Tri-Methyl-Histone H3 (Lys27) [C36B11] Cell Signalling Technology (9733S) Rabbit anti-mouse Di-Methyl-Histone H3 (Lys4) [C64G9] Cell Signalling Technology (9725S) Normal Rabbit IgG (ChIP graded) Cell Signalling Technology (2729) Antibodies and their corresponding dilutions used in Flow Cytometry. .. Rat anti-mouse Sca1 (Ly-6A/E) [E13-161.7] STEMCELL Technology (60032) 1/100 Rat anti-mouse IgG2a, kappa Isotype [RTK2758] STEMCELL Technology (60076) 1/100 Rabbit Anti-mouse/rat S100β [EP1576Y] Abcam (ab52642) 1/100 Normal Rabbit IgG (F graded) Cell Signalling Technology (2729) 1/100 Alexa Fluor 647 Goat anti-mouse (H+L) Life Technologies (A- A-21235) 1/100 Validation For validation, we introduced background blocking controls as well as specificity controls that included secondary antibodies alone.
other:
Whole Genome Amplification:Article Title: Ergothioneine improves healthspan of aged animals by enhancing cGPDH activity through CSE-dependent persulfidation.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-4-HNE, rabbit polyclonal Invitrogen Cat# BS-6313R; RRID: AB_2827741 anti-SIRT1, rabbit polyclonal Proteintech Cat# 13161-1-AP; RRID: AB_10646436 anti-TOM20, rabbit polyclonal Proteintech Cat# 11802-1-AP; RRID: AB_2207530 anti-GPD1, rabbit polyclonal Proteintech Cat# 13451-1-AP; RRID: AB_10646471 anti-PGC1 alpha, rabbit polyclonal Novus Biologicals Cat# NBP1-04676; RRID: AB_1522118 anti-VEGFR2, mouse monoclonal Santa Cruz Biotechnology Cat# sc-6251; RRID: AB_628431 anti-GAPDH, mouse monoclonal Santa Cruz Biotechnology Cat# sc-47724; RRID: AB_627678 anti-PAX-7, mouse monoclonal Santa Cruz Biotechnology Cat# sc-81648; RRID: AB_2159836 WGA-AF488 Invitrogen Cat# W11261 Anti-Ac-Lysine, mouse monoclonal Santa Cruz Biotechnology Cat# sc-32268; RRID: AB_627898 anti- b-tubulin Sigma Aldrich Cat# T0198; RRID: AB_477556 anti-CSE Proteintech Cat# 12217-1-AP; RRID: AB_2087497 anti-Vinculin Invitrogen Cat# MA5-11690; RRID: AB_10976821 anti-S6 Cell Signaling Technology Cat# 2217; RRID: AB_331355 anti-pS6 Cell Signaling Technology Cat# 2211; RRID: AB_331679 anti-mouse-AF488, goat polyclonal Abcam Cat# ab150113; RRID: AB_2576208 anti-mouse-AF680, goat polyclonal Abcam Cat# ab175775 anti-rabbit-AF680, donkey polyclonal Abcam Cat# ab175772 anti-rabbit-CY5, goat polyclonal Invitrogen Cat# A10523; RRID: AB_2534032 anti-rabbit-HRP Abcam Cat# ab205718; RRID: AB_2819160 anti-mouse-HRP Cell Signaling Technology Cat# 7076S;RRID: AB_330924 Bacterial and virus strains OP50-1 Caenorhabditis Genetics Center N/A Chemicals, peptides, and recombinant proteins Chelex-100 Sigma Aldrich Cat# C7901 HEPES VWR Cat# 441485H 1M HEPES solution Sigma Aldrich Cat# H0887 EDTA Sigma Aldrich Cat# E5134 Neocuproine Sigma Aldrich Cat# N1501 IGEPAL Thermofisher Cat# J61055 SDS Carl-Roth Cat# 2326.2 TRIS Carl-Roth Cat# 5429.3 Sodium deoxycholate Sigma Aldrich Cat# D6750 Glycine Carl-Roth Cat# 3187.5 Ponceau S Sigma Aldrich Cat# P3504 Tween Sigma Aldrich Cat# P9416 Triton X-100 Sigma Aldrich Cat# T8787 Protease inhibitor cocktail Sigma Aldrich Cat# P8340 4-Chloro-7-nitrobenzofurazan (NBF-Cl) Sigma Aldrich Cat# 163260 1,4-dithiothreitol (DTT) ThermoFisher Cat# R0861 Iodoacetamide Sigma-Aldrich Cat# I1149 Ammonium bicarbonate Sigma Aldrich Cat# 09830 Calcium chloride Euromedex Cat# T885-A Guanidine hydrochloride Sigma Aldrich Cat# G3272 (Continued on next page) e1 Cell Metabolism 37, 1–15.e1–e14, February 4, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Triethylammonium bicarbonate buffer Sigma Aldrich Cat# 18597 Hydroxylamine solution Sigma Aldrich Cat# 467804 Trypsin Promega Cat# V5117 TMT10plex Thermo Scientific Cat# 90110 DCP-Bio1 Merck Cat# NS1226 Copper (II)-TBTA complex Lumiprobe Cat# 21050 L-ascorbic acid Sigma-Aldrich Cat# A92902 Formic acid Suprapur Supelco Cat# 111670 TCEP Sigma-Aldrich Cat# 646547 AQC Synchem Cat# s041 AccQ-Tag Ultra Borate Buffer Waters Cat# 186009283 Cyanine5 alkyne Lumirpobe Cat# A30B0 Laemmli (4X) buffer BioRad Cat# 1610747 b-mercaptoethanol Sigma Aldrich Cat# M3148 Western Blotting Luminol Reagent Santa Cruz Biotechnology Cat# sc-2048 Ponceau S Sigma Aldrich Cat# P3504 Hydrogen Peroxide Solution Sigma Aldrich Cat# 216763 Erastin Sigma Aldrich Cat# E7781 1S,3R-RSL 3 Sigma Aldrich Cat# SML2234 Thiazolyl Blue Tetrazolium Bromide Sigma Aldrich Cat# M5655 Methyl viologen dichloride hydrate Sigma Aldrich Cat# 856177 Sodium (meta)arsenite Santa Cruz Biotechnology Cat# sc-250986 Nile Red Sigma Aldrich Cat# 72485 Oil Red O Sigma Aldrich Cat# O0625 Bovine Serum Albumin Sigma Aldrich Cat# A7906 L-(+)-Ergothioneine Thermo Scientific Cat# J67861.03 L-Hercynine Sigma Aldrich Cat# SMB00978 DMEM Gibco Cat# 21969-035 DMEM, high glucose, no glutamine, no methionine, no cystine Gibco Cat# 21969-035 DMEM/F-12 Gibco Cat# 21041025 DMEM, no phenol-red Gibco Cat# 31053028 Endothelial Cell Growth Medium 2 (ECGM2) PromoCell Cat# C-22011 Fetal bovine serum Pan Biotech Cat# P30-3031 L-glutamine Pan Biotech Cat# P64-80100 Penicillin-streptomycin Pan Biotech Cat# P66-07100 Polybrene Santa Cruz Biotechnology Cat# 28728-55-4 Lipofectamine RNAiMAX Invitrogen Cat# 13778075 DPBS Sigma Aldrich Cat# D8537 Hoechst 33342 Sigma Aldrich Cat# B2261 L-Cystine Sigma Aldrich Cat# 30200 Pierce High Capacity NeutrAvidin Agarose beads Thermofisher Cat# 20357 Pierce NeutrAvidin Agarose beads Thermofisher Cat# 29200 DTAB Thermo Scientific Cat# A10761.09 Sodium disulfide Dojindo Molecular Technologies Cat# SB-02 Dihydroxyacetone phosphate hemimagnesium salt hydrate Sigma Aldrich Cat# 51269 (Continued on next page) Cell Metabolism 37, 1–15.e1–e14, February 4, 2025 e2
Virus:Article Title: Ergothioneine improves healthspan of aged animals by enhancing cGPDH activity through CSE-dependent persulfidation.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-4-HNE, rabbit polyclonal Invitrogen Cat# BS-6313R; RRID: AB_2827741 anti-SIRT1, rabbit polyclonal Proteintech Cat# 13161-1-AP; RRID: AB_10646436 anti-TOM20, rabbit polyclonal Proteintech Cat# 11802-1-AP; RRID: AB_2207530 anti-GPD1, rabbit polyclonal Proteintech Cat# 13451-1-AP; RRID: AB_10646471 anti-PGC1 alpha, rabbit polyclonal Novus Biologicals Cat# NBP1-04676; RRID: AB_1522118 anti-VEGFR2, mouse monoclonal Santa Cruz Biotechnology Cat# sc-6251; RRID: AB_628431 anti-GAPDH, mouse monoclonal Santa Cruz Biotechnology Cat# sc-47724; RRID: AB_627678 anti-PAX-7, mouse monoclonal Santa Cruz Biotechnology Cat# sc-81648; RRID: AB_2159836 WGA-AF488 Invitrogen Cat# W11261 Anti-Ac-Lysine, mouse monoclonal Santa Cruz Biotechnology Cat# sc-32268; RRID: AB_627898 anti- b-tubulin Sigma Aldrich Cat# T0198; RRID: AB_477556 anti-CSE Proteintech Cat# 12217-1-AP; RRID: AB_2087497 anti-Vinculin Invitrogen Cat# MA5-11690; RRID: AB_10976821 anti-S6 Cell Signaling Technology Cat# 2217; RRID: AB_331355 anti-pS6 Cell Signaling Technology Cat# 2211; RRID: AB_331679 anti-mouse-AF488, goat polyclonal Abcam Cat# ab150113; RRID: AB_2576208 anti-mouse-AF680, goat polyclonal Abcam Cat# ab175775 anti-rabbit-AF680, donkey polyclonal Abcam Cat# ab175772 anti-rabbit-CY5, goat polyclonal Invitrogen Cat# A10523; RRID: AB_2534032 anti-rabbit-HRP Abcam Cat# ab205718; RRID: AB_2819160 anti-mouse-HRP Cell Signaling Technology Cat# 7076S;RRID: AB_330924 Bacterial and virus strains OP50-1 Caenorhabditis Genetics Center N/A Chemicals, peptides, and recombinant proteins Chelex-100 Sigma Aldrich Cat# C7901 HEPES VWR Cat# 441485H 1M HEPES solution Sigma Aldrich Cat# H0887 EDTA Sigma Aldrich Cat# E5134 Neocuproine Sigma Aldrich Cat# N1501 IGEPAL Thermofisher Cat# J61055 SDS Carl-Roth Cat# 2326.2 TRIS Carl-Roth Cat# 5429.3 Sodium deoxycholate Sigma Aldrich Cat# D6750 Glycine Carl-Roth Cat# 3187.5 Ponceau S Sigma Aldrich Cat# P3504 Tween Sigma Aldrich Cat# P9416 Triton X-100 Sigma Aldrich Cat# T8787 Protease inhibitor cocktail Sigma Aldrich Cat# P8340 4-Chloro-7-nitrobenzofurazan (NBF-Cl) Sigma Aldrich Cat# 163260 1,4-dithiothreitol (DTT) ThermoFisher Cat# R0861 Iodoacetamide Sigma-Aldrich Cat# I1149 Ammonium bicarbonate Sigma Aldrich Cat# 09830 Calcium chloride Euromedex Cat# T885-A Guanidine hydrochloride Sigma Aldrich Cat# G3272 (Continued on next page) e1 Cell Metabolism 37, 1–15.e1–e14, February 4, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Triethylammonium bicarbonate buffer Sigma Aldrich Cat# 18597 Hydroxylamine solution Sigma Aldrich Cat# 467804 Trypsin Promega Cat# V5117 TMT10plex Thermo Scientific Cat# 90110 DCP-Bio1 Merck Cat# NS1226 Copper (II)-TBTA complex Lumiprobe Cat# 21050 L-ascorbic acid Sigma-Aldrich Cat# A92902 Formic acid Suprapur Supelco Cat# 111670 TCEP Sigma-Aldrich Cat# 646547 AQC Synchem Cat# s041 AccQ-Tag Ultra Borate Buffer Waters Cat# 186009283 Cyanine5 alkyne Lumirpobe Cat# A30B0 Laemmli (4X) buffer BioRad Cat# 1610747 b-mercaptoethanol Sigma Aldrich Cat# M3148 Western Blotting Luminol Reagent Santa Cruz Biotechnology Cat# sc-2048 Ponceau S Sigma Aldrich Cat# P3504 Hydrogen Peroxide Solution Sigma Aldrich Cat# 216763 Erastin Sigma Aldrich Cat# E7781 1S,3R-RSL 3 Sigma Aldrich Cat# SML2234 Thiazolyl Blue Tetrazolium Bromide Sigma Aldrich Cat# M5655 Methyl viologen dichloride hydrate Sigma Aldrich Cat# 856177 Sodium (meta)arsenite Santa Cruz Biotechnology Cat# sc-250986 Nile Red Sigma Aldrich Cat# 72485 Oil Red O Sigma Aldrich Cat# O0625 Bovine Serum Albumin Sigma Aldrich Cat# A7906 L-(+)-Ergothioneine Thermo Scientific Cat# J67861.03 L-Hercynine Sigma Aldrich Cat# SMB00978 DMEM Gibco Cat# 21969-035 DMEM, high glucose, no glutamine, no methionine, no cystine Gibco Cat# 21969-035 DMEM/F-12 Gibco Cat# 21041025 DMEM, no phenol-red Gibco Cat# 31053028 Endothelial Cell Growth Medium 2 (ECGM2) PromoCell Cat# C-22011 Fetal bovine serum Pan Biotech Cat# P30-3031 L-glutamine Pan Biotech Cat# P64-80100 Penicillin-streptomycin Pan Biotech Cat# P66-07100 Polybrene Santa Cruz Biotechnology Cat# 28728-55-4 Lipofectamine RNAiMAX Invitrogen Cat# 13778075 DPBS Sigma Aldrich Cat# D8537 Hoechst 33342 Sigma Aldrich Cat# B2261 L-Cystine Sigma Aldrich Cat# 30200 Pierce High Capacity NeutrAvidin Agarose beads Thermofisher Cat# 20357 Pierce NeutrAvidin Agarose beads Thermofisher Cat# 29200 DTAB Thermo Scientific Cat# A10761.09 Sodium disulfide Dojindo Molecular Technologies Cat# SB-02 Dihydroxyacetone phosphate hemimagnesium salt hydrate Sigma Aldrich Cat# 51269 (Continued on next page) Cell Metabolism 37, 1–15.e1–e14, February 4, 2025 e2
Protease Inhibitor:Article Title: Ergothioneine improves healthspan of aged animals by enhancing cGPDH activity through CSE-dependent persulfidation.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-4-HNE, rabbit polyclonal Invitrogen Cat# BS-6313R; RRID: AB_2827741 anti-SIRT1, rabbit polyclonal Proteintech Cat# 13161-1-AP; RRID: AB_10646436 anti-TOM20, rabbit polyclonal Proteintech Cat# 11802-1-AP; RRID: AB_2207530 anti-GPD1, rabbit polyclonal Proteintech Cat# 13451-1-AP; RRID: AB_10646471 anti-PGC1 alpha, rabbit polyclonal Novus Biologicals Cat# NBP1-04676; RRID: AB_1522118 anti-VEGFR2, mouse monoclonal Santa Cruz Biotechnology Cat# sc-6251; RRID: AB_628431 anti-GAPDH, mouse monoclonal Santa Cruz Biotechnology Cat# sc-47724; RRID: AB_627678 anti-PAX-7, mouse monoclonal Santa Cruz Biotechnology Cat# sc-81648; RRID: AB_2159836 WGA-AF488 Invitrogen Cat# W11261 Anti-Ac-Lysine, mouse monoclonal Santa Cruz Biotechnology Cat# sc-32268; RRID: AB_627898 anti- b-tubulin Sigma Aldrich Cat# T0198; RRID: AB_477556 anti-CSE Proteintech Cat# 12217-1-AP; RRID: AB_2087497 anti-Vinculin Invitrogen Cat# MA5-11690; RRID: AB_10976821 anti-S6 Cell Signaling Technology Cat# 2217; RRID: AB_331355 anti-pS6 Cell Signaling Technology Cat# 2211; RRID: AB_331679 anti-mouse-AF488, goat polyclonal Abcam Cat# ab150113; RRID: AB_2576208 anti-mouse-AF680, goat polyclonal Abcam Cat# ab175775 anti-rabbit-AF680, donkey polyclonal Abcam Cat# ab175772 anti-rabbit-CY5, goat polyclonal Invitrogen Cat# A10523; RRID: AB_2534032 anti-rabbit-HRP Abcam Cat# ab205718; RRID: AB_2819160 anti-mouse-HRP Cell Signaling Technology Cat# 7076S;RRID: AB_330924 Bacterial and virus strains OP50-1 Caenorhabditis Genetics Center N/A Chemicals, peptides, and recombinant proteins Chelex-100 Sigma Aldrich Cat# C7901 HEPES VWR Cat# 441485H 1M HEPES solution Sigma Aldrich Cat# H0887 EDTA Sigma Aldrich Cat# E5134 Neocuproine Sigma Aldrich Cat# N1501 IGEPAL Thermofisher Cat# J61055 SDS Carl-Roth Cat# 2326.2 TRIS Carl-Roth Cat# 5429.3 Sodium deoxycholate Sigma Aldrich Cat# D6750 Glycine Carl-Roth Cat# 3187.5 Ponceau S Sigma Aldrich Cat# P3504 Tween Sigma Aldrich Cat# P9416 Triton X-100 Sigma Aldrich Cat# T8787 Protease inhibitor cocktail Sigma Aldrich Cat# P8340 4-Chloro-7-nitrobenzofurazan (NBF-Cl) Sigma Aldrich Cat# 163260 1,4-dithiothreitol (DTT) ThermoFisher Cat# R0861 Iodoacetamide Sigma-Aldrich Cat# I1149 Ammonium bicarbonate Sigma Aldrich Cat# 09830 Calcium chloride Euromedex Cat# T885-A Guanidine hydrochloride Sigma Aldrich Cat# G3272 (Continued on next page) e1 Cell Metabolism 37, 1–15.e1–e14, February 4, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Triethylammonium bicarbonate buffer Sigma Aldrich Cat# 18597 Hydroxylamine solution Sigma Aldrich Cat# 467804 Trypsin Promega Cat# V5117 TMT10plex Thermo Scientific Cat# 90110 DCP-Bio1 Merck Cat# NS1226 Copper (II)-TBTA complex Lumiprobe Cat# 21050 L-ascorbic acid Sigma-Aldrich Cat# A92902 Formic acid Suprapur Supelco Cat# 111670 TCEP Sigma-Aldrich Cat# 646547 AQC Synchem Cat# s041 AccQ-Tag Ultra Borate Buffer Waters Cat# 186009283 Cyanine5 alkyne Lumirpobe Cat# A30B0 Laemmli (4X) buffer BioRad Cat# 1610747 b-mercaptoethanol Sigma Aldrich Cat# M3148 Western Blotting Luminol Reagent Santa Cruz Biotechnology Cat# sc-2048 Ponceau S Sigma Aldrich Cat# P3504 Hydrogen Peroxide Solution Sigma Aldrich Cat# 216763 Erastin Sigma Aldrich Cat# E7781 1S,3R-RSL 3 Sigma Aldrich Cat# SML2234 Thiazolyl Blue Tetrazolium Bromide Sigma Aldrich Cat# M5655 Methyl viologen dichloride hydrate Sigma Aldrich Cat# 856177 Sodium (meta)arsenite Santa Cruz Biotechnology Cat# sc-250986 Nile Red Sigma Aldrich Cat# 72485 Oil Red O Sigma Aldrich Cat# O0625 Bovine Serum Albumin Sigma Aldrich Cat# A7906 L-(+)-Ergothioneine Thermo Scientific Cat# J67861.03 L-Hercynine Sigma Aldrich Cat# SMB00978 DMEM Gibco Cat# 21969-035 DMEM, high glucose, no glutamine, no methionine, no cystine Gibco Cat# 21969-035 DMEM/F-12 Gibco Cat# 21041025 DMEM, no phenol-red Gibco Cat# 31053028 Endothelial Cell Growth Medium 2 (ECGM2) PromoCell Cat# C-22011 Fetal bovine serum Pan Biotech Cat# P30-3031 L-glutamine Pan Biotech Cat# P64-80100 Penicillin-streptomycin Pan Biotech Cat# P66-07100 Polybrene Santa Cruz Biotechnology Cat# 28728-55-4 Lipofectamine RNAiMAX Invitrogen Cat# 13778075 DPBS Sigma Aldrich Cat# D8537 Hoechst 33342 Sigma Aldrich Cat# B2261 L-Cystine Sigma Aldrich Cat# 30200 Pierce High Capacity NeutrAvidin Agarose beads Thermofisher Cat# 20357 Pierce NeutrAvidin Agarose beads Thermofisher Cat# 29200 DTAB Thermo Scientific Cat# A10761.09 Sodium disulfide Dojindo Molecular Technologies Cat# SB-02 Dihydroxyacetone phosphate hemimagnesium salt hydrate Sigma Aldrich Cat# 51269 (Continued on next page) Cell Metabolism 37, 1–15.e1–e14, February 4, 2025 e2
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