Cultured U87-MG cells were lysed in lysis buffer (20 mM pH 7.5 Tris, 150 mM NaCl, 1% TritonX-100, 1 mM EDTA and protease inhibitor cocktail). The lysates were cleared by centrifugation and subsequently incubated with either 20 μg of IgG or 2 μg of LAMP-2A antibody(Abcam, 125068), or 2 μg of Hsc70 antibody(Proteintech, 10654-1-AP) for 2 h with rotation on ice. Protein G-Agarose beads (Roche, Mannheim, Germany) were then added followed by overnight rotation at 4 °C. Immunoprecipitants were separated on SDS-PAGE gel. LAMP-2A(1:1000, Abcam, 125068), Hsc70 (1:3000, Proteintech, 66442-1-Ig) and N-CoR (1:500, Abcam, ab3482) were detected by western blotting as previously mentioned.
Lamp 2a antibody
Manufactured by Abcam
Sourced in United Kingdom
The LAMP-2A antibody is a protein-specific antibody that recognizes the LAMP-2A (Lysosome-associated membrane glycoprotein 2A) protein. LAMP-2A is a key component of the chaperone-mediated autophagy pathway, which is involved in the selective degradation of cytosolic proteins in lysosomes. The antibody can be used to detect and study the expression and localization of LAMP-2A in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Ab3482, by Abcam (1 mentions)
Protein g agarose beads, by Roche (1 mentions)
Cycloheximide (chx), by Cell Signaling Technology (1 mentions)
Secondary antibodies conjugated to horseradish peroxidase, by Santa Cruz Biotechnology (1 mentions)
Anti gapdh, by Santa Cruz Biotechnology (1 mentions)
Bortezomib ps 341, by Selleck Chemicals (1 mentions)
Goat polyclonal anti cox 2, by Santa Cruz Biotechnology (1 mentions)
Nrf2 sirna, by Santa Cruz Biotechnology (1 mentions)
Control shrna, by Santa Cruz Biotechnology (1 mentions)
Control sirna, by Santa Cruz Biotechnology (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Tnf α, by R&D Systems (1 mentions)
2 protocols using lamp 2a antibody
1
Immunoprecipitation of LAMP-2A and Hsc70
2
Molecular Mechanisms in Cell Signaling
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Rabbit polyclonal anti-IκBα, anti-LC3B, anti-phospho-mTOR (Ser2448), anti-sirtuin1, and anti-KEAP1 antibodies, cycloheximide, p65 siRNAs, and LC3B siRNAs were purchased from Cell Signaling Technology (Danvers, USA). Rabbit polyclonal anti-p65, anti-cIAP2, anti-PARP, anti-Nrf2(C-20), anti-Lamin A/C, goat polyclonal anti-COX-2, anti-GAPDH, mouse monoclonal Lamp2 antibodies, secondary antibodies conjugated to horseradish peroxidase, Nrf2 siRNAs, control siRNAs, Lamp2 shRNAs, and control shRNAs were obtained from Santa Cruz Biotechnology (Santa Cruz, USA). Rabbit polyclonal Lamp2a antibody was from Abcam (Cambridge, UK). TNF-α was from R&D Systems (Minneapolis, USA). Bortezomib (PS-341) was obtained from Selleckchem (Houston, USA) and MG132 was from Calbiochem (Darmstadt, Germany). The eutomer of dehydroxymethylepoxyquinomicin (DHMEQ), (−)-DHMEQ, was from ChemScene (Monmouth Junction, USA). Lipofectamine 2000 was purchased from Invitrogen (Carlsbad, USA). 3-MA and thiazolyl blue tetrazolium blue (MTT) was from Sigma-Aldrich, Inc. (St. Louis, USA).
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