resource source identifier antibodies glut1  (Alomone Labs)

Journal: Cell reports methods

Article Title: In situ microwave fixation provides an instantaneous snapshot of the brain metabolome.

doi: 10.1016/j.crmeth.2023.100455

Figure Lengend Snippet: Figure 5. Immunofluorescent analysis of GLUT2 and PC expression in CTX, HIPP, and BS in a mouse model of T1DM (A–C) 20 mm coronal sections of the mouse brain were stained with GLUT2 (green) and NeuN (red) and quantified in in the CTX (A), HIPP (B), and brainstem (BS) (C). Zoomed-in images are shown for the retro-splenial (RS) cortex, Cornu Ammonis 3 (CA3) of HIPP, and the DVC of BS. (D) Quantification of % positive pixels for GLUT2 using HALO software. (E–G) 20 mm coronal sections of the mouse brain were stained with pyruvate carboxylase (PC) (green) and NeuN (red) and quantified in in the CTX (E), HIPP (F), and BS (G). Zoomed-in images are shown for the RS, the CA3 of HIPP, and the DVC of BS. (H) Quantification of percentage of positive pixels for PC using HALO software. Values are presented as mean ± SEM (n = 4 biological replicates), *p < 0.05, **p < 0.01, ***p < 0.001, analyzed by Student’s t test. Scale bars are either 1 mm, 500 mm, or 100 mm. PC, pyruvate carboxylase; GLUT2, glucose transporter 2.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies GLUT1, Rabbit Abcam Cat# ab115730; RRID: AB_10903230 GLUT2, Rabbit Novus Cat# NBP2-22218; RRID: AB_2335858 GLUT3, Rabbit Alomone Cat# AGT-023; RRID:AB_2756644 Pyruvate Carboxylase, Rabbit GeneTex GTX121987 Pyruvate Dehydrogenase (ser 293), Rabbit EMD Milipore Cat# AP1062; RRID:AB_2863934 NeuN, Mouse Abcam ab279295 NeuN, Rabbit Abcam Cat# ab236870; RRID:AB_2927651 GFAP, Chicken GeneTex Cat# GTX85454; RRID:AB_10621124 DAPI Novus NBP2-31156 Goat anti-chicken, Alexa 594 Thermo Fisher Scientific Cat# A-11042; RRID:AB_2534099 Goat anti-rabbit, Alexa 488 Thermo Fisher Scientific Cat# A-11034; RRID:AB_2576217 Goat anti-mouse, Alexa 647 Thermo Fisher Scientific Cat# A-21236; RRID:AB_2535805 Chemicals, peptides, and recombinant proteins U-13C6-glucose, 99% Cambridge Isotope Laboratories CLM-1396-MPT-PK L-Norvaline Sigma-Aldrich N7627-5G Methoxyamine hydrochloride Sigma-Aldrich 226904-25G Pyridine Thermo Fisher Scientific TS-27530 MSTFA +1 Percent TMCS Sigma-Aldrich 375934-10X1ML Hydrochloric Acid Thermo Fisher Scientific HPLC Methanol Thermo Fisher Scientific 34860-4X4L-R PBS Sigma-Aldrich P3813 Heparin Sodium (10,000units/mL) N/A NDC# 25021040010 4% Paraformaldehyde in PBS Thermo Fisher Scientific J19943.K2 Sucrose Thermo Fisher Scientific 036508.A1 Normal Goat Serum Thermo Fisher Scientific 31873 Triton X- Sigma-Aldrich X100 Tween 20 Sigma-Aldrich P1379 Prolong Glass Thermo Fisher Scientific P36984 Streptozotocin Sigma-Aldrich 572201 Citric Acid Thermo Fisher Scientific A940 Sodium citrate dihydrate Thermo Fisher Scientific BP327 2-methylbutane Thermo Fisher Scientific 019387AP Ethanol Decon Labs CAS#: 64-17-5 OCT compound Thermo Fisher Scientific 23-730-571 Deposited data Raw and processed metabolomics data This study Metabolomics Workbench ST002524 and ST002525 Software and algorithms HALO Software Indica Labs N/A Prism Graphpad N/A IsoCorrectoR Heinrich et al.80 N/A DEXSI software Dagley and McConville81 N/A Metaboanalyst Xia et al.82 (https://chemdata.nist.gov/dokuwiki/doku. php?id=chemdata:amdis) (Continued on next page) e1 Cell Reports Methods 3, 100455, April 24, 2023

Techniques: Expressing, Staining, Software