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3 protocols using anti flag sab4200071

1

Antibody Validation for Cell Signaling

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Anti-MYC (2276), anti-GAPDH (2118), and anti-LC3A/B (4108) were purchased from Cell Signaling Technology. Anti-FLAG (SAB4200071) was purchased from Sigma-Aldrich. Anti-HA (ab18181) and anti-USP15 (ab97533) were purchased from Abcam. Lipopolysaccharide (LPS; serotype 0128: B12), chloroquine (CQ; C6628), dimethyl sulfoxide (DMSO; 472301), puromycin (P8833), paraformaldehyde (P6148), Triton X-100 (T8787), 3-methyladenine (3-MA; M9281), gentamicin (G1272), deoxycholate (D6750), and Dulbecco’s phosphate-buffered saline (DPBS; D8537) were purchased from Sigma-Aldrich. Lipofectamine 2000 (11668019) was purchased from Thermo Fisher Scientific.
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2

Optimized Synthesis and Characterization of LM9

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LM9 was synthesized with a purity over 97% in our laboratory. PA was solubilized in low-endotoxin bovine serum albumin (BSA) (5%) at a final concentration of 5 mM. PA and BSA were purchased from Sigma-Aldrich (St. Louis, MO, USA). The MTT assay kit and oil red O staining kit (for cultured cells) were acquired from Solarbio (Beijing, China). Antibodies were purchased from the following suppliers: anti-lamin B1 (ab133741), anti-IκB alpha (ab133462), anti-Ly6G (ab25377), anti-TNF-α (ab6671), anti-collagen I (ab6308), and anti-collagen IV (ab6586) were obtained from Abcam (Cambridge, MA, USA). Anti-TLR4 (sc-293072) and anti-TGF-β (sc-130348) were obtained from Santa Cruz (Dallas, TX, USA). Anti-FLAG (SAB4200071) and anti-HA (MFCD00803873) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-ICAM-1 (Cat# 60299-1-Ig) was obtained from Proteintech (Wuhan, China). Two plasmids encoding pCMV-HA-MyD88 and Flag-MyD88 were obtained from Sino Biological Inc. (Beijing, China).
ELISA kits for mouse TNF-α, IL-1β, and IL-6 were purchased from Invitrogen (Carlsbad, CA, USA). Nuclear and cytoplasmic protein extraction kits were from KeyGEN Biotech (Nanjing, China). PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) was purchased from Takara Bio (RR047A, Shiga, Japan).
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3

Antibody Sourcing and Cell Treatments

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Anti-Myc (2276), anti-GAPDH (2118), and anti-LC3A/B (4108) were purchased from Cell Signaling Technology (MA, USA). Anti-Flag (SAB4200071) was purchased from Sigma-Aldrich (St Louis, MO, USA). Anti-HA and anti-HBx antibodies were purchased from Abcam (Cambridge, MA). Lipopolysaccharide (LPS; serotype 0128:B12), chloroquine (CQ; C6628), dimethyl sulfoxide (DMSO; 472301), puromycin (P8833), paraformaldehyde (P6148), Triton X-100 (T8787), 3-methyladenine (3-MA;M9281), gentamicin (G1272), deoxycholate (D6750), and Dulbecco's phosphate-buffered saline (DPBS; D8537) were purchased from Sigma-Aldrich. Lipofectamine 2000 (11668019) was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
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