The AGS3 polyclonal antibody has been described previously and was kindly provided Dr. D. Ma (15 (link)). The commercial antibodies used as follows: LC3B antibody (L7543), β-actin-peroxidase antibody (A3854) [Sigma-Aldrich]; LAMP-1 (1D4B), LAMP-2 (H4B4), and CD-63/LAMP-3 (H5C6) antibodies [Developmental Studies Hybridoma Bank]; TGN-46 (ab16059) and LAMP-1 (ab25630) [Abcam]; cathepsin D (sc-6486) [Santa Cruz]; Akt (11E7), phospho-Akt (Ser473) (D9E), TSC2 (D93F12), phospho-TSC2 (Thr 1462) (5B12), 4E-BP1 (53H11), phospho-4E-BP1 (Thr37/46) (236B4), p70 S6 kinase (49D7), phospho-p70 S6 kinase (Thr389) (108D2) and GAPDH (14C10) [Cell Signaling]; and TFEB antibody (A303-672A) [Bethyl Laboratories]. The following reagents were used phorbol12-myristate 13-acetate (PMA) [Sigma-Aldrich]; bafilomycin A1 and glycogen synthase kinase 3 (GSK3) inhibitor VIII [Calbiochem]; G418 sulfate [Cellgro]; lipopolysaccharide (LPS) [Enzo Life Sciences]; Staphylococcus aureus (Wood strain without protein A) BioParticles, Escherichia coli (K-12 strain) BioParticles and DQ-Red BSA [Molecular Probes].
Protocol detail
Antibody Reagents for Autophagy Research
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4e bp1
Acetate
Actin
Antibodies
Antibody
Bafilomycin a1
Cathepsin d
Escherichia coli
G418
Gapdh
Glycogen synthase kinase 3
Hybridoma
Lamp 1
Lamp 2
Lamp 3
Lipopolysaccharide
Molecular probes
Myristate
P70 s6 kinase
Peroxidase
Protein a
Staphylococcus aureus
Strain
Sulfate
Tfeb
Tsc2
Corresponding Organization : National Institutes of Health
Other organizations : University of Maryland, College Park, Medical University of South Carolina
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Variable analysis
independent variables
- Phorbol12-myristate 13-acetate (PMA)
- Bafilomycin A1
- Glycogen synthase kinase 3 (GSK3) inhibitor VIII
- Lipopolysaccharide (LPS)
- Staphylococcus aureus (Wood strain without protein A) BioParticles
- Escherichia coli (K-12 strain) BioParticles
- DQ-Red BSA
- Protein expression (e.g., LC3B, β-actin, LAMP-1, LAMP-2, CD-63/LAMP-3, TGN-46, cathepsin D, Akt, phospho-Akt, TSC2, phospho-TSC2, 4E-BP1, phospho-4E-BP1, p70 S6 kinase, phospho-p70 S6 kinase, GAPDH, TFEB)
- AGS3 polyclonal antibody
- Commercial antibodies (LC3B, β-actin-peroxidase, LAMP-1, LAMP-2, CD-63/LAMP-3, TGN-46, LAMP-1, cathepsin D, Akt, phospho-Akt, TSC2, phospho-TSC2, 4E-BP1, phospho-4E-BP1, p70 S6 kinase, phospho-p70 S6 kinase, GAPDH, TFEB)
- G418 sulfate
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