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2 protocols using mouse anti actin

1

Antibody Validation for Signaling Pathways

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The following antibodies were purchased from Cell Signaling Technology: rabbit anti-IRS-1 (2382), rabbit anti-pAKT S473 (9271), rabbit anti-p85 (4292), rabbit anti-Grb2 (3972), rabbit anti-FoxO1 (2880), rabbit anti-ERK1/2 (9102), rabbit anti-pERK1/2 (4370), anti-IGF-1Rβ (9750), anti-IRβ (3025), anti-pIGF-1Rβ Y1135/1136/pIRβ Y1150/1151 (3024), anti-pIGF-1Rβ Y1131/pIRβ 1146 (3021). Other antibodies were from the following commercial sources: rabbit anti-pIRS-1 Y612 (Invitrogen, 44-816 G); rabbit anti-AKT (HUABIO, ET1609-47), rabbit anti-GFP (HUABIO, ET1607-31), mouse anti-Actin (HUABIO, M1210-2), mouse anti-MHC (DSHB, MF20), mouse anti-FLAG (YEASEN, 30503ES60), and mouse anti-mCherry (ABclonal, AE002). HRP-conjugated secondary antibodies: goat anti-mouse second antibody (Jackson, 115-035-003) and goat anti-rabbit second antibody (Jackson, 111-035-003). Fluorescent secondary antibodies were from Thermo Fisher Scientific: goat anti-mouse-Alexa Fluor 546 (A-11003), goat anti-rabbit-Alexa Fluor 488 (A-11008), goat anti-rabbit-Alexa Fluor 546 (A-11010), goat anti-rabbit-Alexa Fluor 633 (A-21071).
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2

Antibody Characterization for Cell Analysis

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The antibodies used were as follows: rat anti-Ki67 (#14-5698-82, Thermo Fisher Scientific, 1:500), rabbit anti-Olig2 (#AB9610, Millipore, 1:400), rat anti-MBP (#MCA409S, Bio-Rad Laboratories, 1:500), mouse anti-CC1 (#ab16794, Abcam, 1:200), goat anti-PDGFRα (#AF1062, R&D Systems, 1:100), rabbit anti-PDGFRα (#3164 S, Cell Signaling Technology, 1:1000), mouse anti-P65 (#6956 S, Cell Signaling Technology, 1:1000), mouse anti-p-P65 (#3036 S, Cell Signaling Technology, 1:1000), mouse anti-Actin (#M1210-2, HUABIO, 1:5000), mouse anti-Tubulin (#M1305-2, HUABIO, 1:5000), mouse anti-RIPK1 (#610458, BD, 1:1000), rabbit anti-p-RIPK1 (#65746, Cell Signaling Technology, 1:1000). Secondary antibodies: CyTM3 affinipure donkey anti-rat IgG (H + L) (#712-165-153, 1:400), CyTM3 affinipure donkey anti-mouse IgG (H + L) (#715-165-151, 1:400), Alexa Fluor 488 affinipure donkey anti-rabbit IgG (H + L) (#711-545-152, 1:400) and Alexa Fluor 488 affinipure donkey anti-goat IgG (H + L) (#705-545-003, 1:400) were purchased from Jackson ImmunoResearch; Goat anti-mouse IgG (H + L) conjugated with HRP (#31430, 1:20000), goat anti-rabbit IgG (H + L) conjugated with HRP (#31,460, 1:20000) were purchased from Thermo Fisher Scientific.
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