Antibodies were from the following companies; mouse mAb anti-myostatin (MstnPP) (6H12) (Abcam & ThermoFischer Scientific); goat pAb anti-human myostatin (amino acid residues 268–376) (R & D systems); mouse mAb anti-APP 6E10 against Aβ epitope RHDSGYE (BioLegend); mouse mAb anti-APP 22C11 against the aminoterminal residues 66–81 (Merck Millipore); rabbit pAb anti-Giantin ab24586 (Abcam); rabbit pAb anti-LC3B NB100-2220 (Novus biological); rabbit pAb anti-Lamp1 ab24170 (Abcam); rabbit pAb anti-GRP-78 H-129 (Santa Cruz); rabbit pAb anti-GFP A-6455 (ThermoFischer Scientific); rabbit mAb anti-Calreticulin ERP3924 (Merck Millipore); rabbit pAb anti-Calnexin C4731 (Sigma); rabbit pAb anti-Ubiquitin Z0458 (DAKO); mouse mAb anti-Actin (MP Biomedicals); Alexa Fluor-conjugated secondary antibodies (Molecular Probes); HRP-coupled secondary goat antibodies (Dianova). Chemicals were purchased from Sigma or Roth.
Anti lamp1 ab24170
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-LAMP1 (ab24170) is a rabbit monoclonal antibody that recognizes the LAMP1 protein. LAMP1 is a lysosomal membrane protein that plays a role in lysosome biogenesis and function.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions)
Anti giantin ab24586, by Abcam (1 mentions)
Anti grp 78 h 129, by Santa Cruz Biotechnology (1 mentions)
Anti calnexin c4731, by Merck Group (1 mentions)
Anti ubiquitin z0458, by Agilent Technologies (1 mentions)
Anti actin, by MP Biomedicals (1 mentions)
Anti lc3b nb100 2220, by Novus Biologicals (1 mentions)
Alexa fluor 647 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions)
Cleaved caspase 3 antibody 9661, by Cell Signaling Technology (1 mentions)
Lysotracker dnd99 red, by Thermo Fisher Scientific (1 mentions)
Anti foxp3 fjk 16s, by Thermo Fisher Scientific (1 mentions)
Anti rab11 d4f5, by Cell Signaling Technology (1 mentions)
Anti tcrβ h57 597, by Thermo Fisher Scientific (1 mentions)
Anti cd3 ucht1, by Thermo Fisher Scientific (1 mentions)
Anti cd3ζ 6b10.2, by Santa Cruz Biotechnology (1 mentions)
Cytochalasin d ab143484, by Abcam (1 mentions)
Neurobasal culture medium, by Thermo Fisher Scientific (1 mentions)
Dmem culture medium, by Thermo Fisher Scientific (1 mentions)
Chloroquine cq, by Merck Group (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
9 protocols using anti lamp1 ab24170
1
Comprehensive Antibody Protocol Inventory
2
Comprehensive Immunofluorescence Assay
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Anti-P. aeruginosa (ab68538) and anti-LAMP1 (ab24170) antibodies were obtained from AbCam. LysoTracker-DND99 Red, Alexa conjugated Annexin V, Rhodamine and Alexa-fluor 647 conjugated phalloidin, TO-PRO3 and CellTrace were acquired from Life Technologies. Cleaved Caspase-3 antibody (9661S) was obtained from Cell Signaling Technology. Purified Recombinant Annexin V was either purchased from BD Pharmingen or produced in the laboratory from pProEx.Htb.Annexin V plasmid (kindly provided by Dr. Seamus J. Martin)[63 (link)].
3
Comprehensive T Cell Immunophenotyping
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For the flow cytometry analysis, anti-CD4 (GK1.5), anti-CD8α (53–6.7), anti-CD44 (IM7), anti-CD127 (A7R34), anti-CD69 (FN50), anti-Bcl-2 (BCL/10C4) were purchased from Biolegend. Anti-CD62L (MEL-14), anti-CD69 (H1.2F3), anti-CD5 (53–7.3), anti-CD45.1 (A20), anti-CD45.2 (104), mouse lgG1 κ isotype control (556027), anti-CD25 (PC61), Rat lgG2a isotype control (eBRG1) and purified anti-human-CD3 (OKT3) were purchased from BD Bioscience. Anti-Foxp3 (FJK-16s), anti-TCRβ (H57–597) and anti-CD3 (UCHT1) were purchased from eBioscience. Fluo-4,AM (F14217) was purchased from Invitrogen. Streptavidin (NEBN7021S) was purchased from Sigma.
For western analysis, anti-α-tubulin (sc-53029) and anti-Arp3 (sc15390) antibodies were purchased from Santa Cruz Biotechnology. Anti-Arpc2 (ab96779) was purchased from Abcam.
For immunofluorescence analysis, anti-Mouse TCR β-chain-488 (H57-597) and anti-Mouse CD3-488 (17A2) were purchased from Biolegend. Anti-CD3ζ (6B10.2) was purchased from Santa Cruz. Anti-EEA1 (C45B10), anti-Rab5 (C8B1) and anti-Rab11 (D4F5) were purchased from Cell Signaling Technology. Anti-LAMP1 (ab24170) was purchased from Abcam.
Cytochalasin D (ab143484) was purchased from Abcam. Staphylococcal enterotoxin E (ET404) was purchased from Toxin Technology. CELLTRACKER(TM) BLUE CMAC (C2110) was purchased from Invitrogen.
For western analysis, anti-α-tubulin (sc-53029) and anti-Arp3 (sc15390) antibodies were purchased from Santa Cruz Biotechnology. Anti-Arpc2 (ab96779) was purchased from Abcam.
For immunofluorescence analysis, anti-Mouse TCR β-chain-488 (H57-597) and anti-Mouse CD3-488 (17A2) were purchased from Biolegend. Anti-CD3ζ (6B10.2) was purchased from Santa Cruz. Anti-EEA1 (C45B10), anti-Rab5 (C8B1) and anti-Rab11 (D4F5) were purchased from Cell Signaling Technology. Anti-LAMP1 (ab24170) was purchased from Abcam.
Cytochalasin D (ab143484) was purchased from Abcam. Staphylococcal enterotoxin E (ET404) was purchased from Toxin Technology. CELLTRACKER(TM) BLUE CMAC (C2110) was purchased from Invitrogen.
4
Autophagy Modulation in Neuronal Cells
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The MTT, poly-l -lysine and chloroquine (CQ) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-LC3B (83506S), anti-SQSTM1/P62 (5114T), anti-ATG5 (12994S), Anti-CSTB (31718), anti-p-Akt (2965S), anti-Akt (4691S), anti-p-mTOR (2971S) and anti-mTOR (2972S) were obtained from Cell Signaling Technology. Anti-TFEB (13372-1-AP) was obtained from Proteintech (Rosemount, IL, USA). Anti-LAMP1 (ab24170) was purchased from Abcam (Cambridge, MA, USA). Neurobasal culture medium, fetal bovine serum (FBS), DMEM culture medium, B27, l -glutamine and d -glucose were purchased from Gibco.
5
Western Blot Analysis of Lysosomal Proteins
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Cells were lysed with 1× NuPAGE LDS buffer (Thermo Fischer Scientific) containing 100 mM DTT and were boiled for 10 min. Approximately 15 µg of protein was loaded onto 4–12% Bis-Tris gels (GenScritp). Proteins were transferred to polyvinylidene difluoride membranes and were blocked with 4% skim powdered milk dissolved in Tris-buffered saline with 0.1% Tween (TBSt) buffer. Primary antibodies were incubated overnight at 4°C in 2% BSA in TBSt buffer, and secondary antibodies were incubated at RT in 4% skim milk in TBSt for 1 h. Anti-LAMP1(ab24170) and recombinant anti-ATP6V0D1/P39 (ab202897) antibodies were purchased from Abcam. Anti-LC3B (L7543-100UL) and anti-actin (MAB1501, clone C4; Millipore) were purchased from Sigma-Aldrich. Secondary HRP-linked antibodies were from GE Healthcare. Blots were developed by using peroxidase-based ECL (Pierce) and detected by using a ChemiDoc Imaging System (Bio-Rad).
6
Recombinant TGF-β1 Autophagy Regulation
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Recombinant human TGF-β1 (580704) was obtained from Biolegend (San Diego, CA). CQ (C6628), 3-MA (M9281) and Baf a1 (B1793) were all purchased from Sigma-Aldrich (St. Louis, MO). The anti-LC3B antibody (L7543) was obtained from Sigma-Aldrich. Oil Red O (O9755) was purchased from Sigma-Aldrich. BODIPY 558/564 Red C12 (D3835) and BODIPY 493/503 (D3922) were both obtained from Invitrogen. DAPI (D9542) was obtained from Sigma-Aldrich, and Alexa Flour 488 or 594-conjugated anti-mouse or anti-rabbit IgG were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA). The anti-Beclin-1 antibody (NBP1-00085) was purchased from Novus Biologicals (Littleton, CO). Anti-LAMP1 (ab24170), anti-SQSTM1 (ab56416) and anti- α-SMA (ab124964) antibodies were all obtained from Abcam (Cambridge, MA). Anti-cleaved caspase-3 antibody (#9664) was obtained from Cell signaling Technology (Danvers, MA), and anti-GAPDH antibody (sc-365062) was purchased from Santa Cruz (Santa Cruz, CA).
7
Antibody Sourcing for Cell Research
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Mouse monoclonal antibodies used in this study were purchased from the following companies: anti-CD147 (306202) from BioLegend, anti-MMP1 (sc-58377) from Santa Cruz, anti-GFP (04363-24) from Nacalai tesque, anti-HA (MMS-101P) from Covance Research Products, and anti-Actin (A5441) from Sigma-Aldrich. Rabbit polyclonal anti-TRE17/USP6 (ab179751) and anti-LAMP1 (ab24170) antibodies were purchased from Abcam, and goat polyclonal anti-MMP9 (AF911) antibody was purchased from R&D Systems. All Alexa-Fluor-conjugated secondary antibodies were purchased from Invitrogen Molecular Probes.
8
TDCIPP-Induced Alzheimer's Autophagy Inhibition
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Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) (98% purity) was purchased from Macklin Reagent (Shanghai, China). Chloroquine (CQ) (97% purity) was purchased from MedChemExpress (Shanghai, China) and used here as an autophagy inhibitor. Tandem mass tag (TMT)-labeling kits were purchased from Thermo Scientific (NJ, USA). Sequencing-grade trypsin/Lys-C mix mixtures were purchased from Promega (WI, USA). Human Aβ1-42 (amyloid beta 1–42) ELISA kits were purchased from Elabscience (Wuhan, China). CYTO-ID® Autophagy Detection kit purchased from Enzo Life Sciences (NY, USA). Tricine-SDS‒PAGE Gel Preparation kits were purchased from Sangon Biotech (Shanghai, China). Anti-presenilin-1 (anti-PS1, AF0245) was purchased from Affinity Bioscience (Jiangsu, China). Anti-LC3 (12741T) and anti-BACE1 (5606S) antibodies were purchased from Cell Signaling Technology (MA, USA). Anti-P62 (ab109012), anti-CTSD (ab6313), and anti-LAMP1 (ab24170) antibodies were purchased from Abcam (Cambridge, UK).
9
Antibodies for Cell Signaling Analysis
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Antibodies used in this study were obtained from the following suppliers: anti-cyclin A (1:500 for immunoblotting, sc-596), anti-E2F1 (1:500, sc-251), anti-GST (1:3000, sc-138), and anti-E2F3 (1:500, sc-878 and 1:500, sc-879) were from Santa-Cruz Biotechnology; anti-Flag (1:1000, #F7425 and #F1804) and anti-α-tubulin (1:2000, #T9026) were from Sigma-Aldrich; anti-E1A (1:2000, #554155), anti-Tim23 (1:1000, #611222), and anti-Tom20 (1:700, #612278) were BD Biosciences; anti-p14ARF (1:500, K0084-3) and anti-Atg5 (1:500, PM050) were from MBL Life science; anti-Hsp70 (ADI-SPA-810) was from Enzo Life Sciences; anti-LC3B (1:2000, NB100-2220) was from Novus Biologicals; anti-Drp1 (1:1000, A303-410A) was from Bethyl Laboratories; and anti-LAMP1 (ab24170) was from Abcam. An anti-E2F3d polyclonal antibody against a peptide, PQPTSYSRLRTK, near the C-terminus was generated in rabbits and affinity-purified (1:100, Eurofins). BFA was purchased from Tocris Bioscience.
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