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Anti lamp1

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Anti-LAMP1 is a laboratory reagent used to detect the presence of the LAMP1 protein in biological samples. LAMP1 is a lysosomal membrane protein that serves as a marker for lysosomes. The Anti-LAMP1 reagent can be used in various analytical techniques, such as immunohistochemistry and flow cytometry, to study lysosomal biology and function.

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Lab products found in correlation

Anti eea1, by BD (2 mentions) Lsm 880 confocal microscope, by Zeiss (2 mentions) F3165, by Miltenyi Biotec (2 mentions) F7425, by Miltenyi Biotec (2 mentions) Anti cd63, by Santa Cruz Biotechnology (2 mentions) Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (2 mentions) Alexa fluor 555 goat anti mouse igg, by Thermo Fisher Scientific (2 mentions) Anti sqstm1 p62, by Abcam (2 mentions) Anti mouse igg hrp, by Cell Signaling Technology (2 mentions) Anti dfcp1 zfyve1, by ABclonal (2 mentions) Anti α synuclein, by BD (2 mentions) Anti β actin, by Bioworld Technology (2 mentions) Anti gapdh, by Bioworld Technology (2 mentions) Anti rabbit igg hrp, by Cell Signaling Technology (2 mentions) Anti c myc, by Santa Cruz Biotechnology (2 mentions) Anti gfap, by Cell Signaling Technology (2 mentions) Anti rab7 d95f2, by Cell Signaling Technology (2 mentions) Mabt837, by Merck Group (2 mentions) Lsm 710 axioobserver confocal microscope, by Zeiss (2 mentions) Fluoromount g, by Southern Biotech (2 mentions)

Close spelling variants of this product

LAMP1 (24 citations) Anti-LAMP1 antibody (5 citations) Rabbit anti-Lamp1 (3 citations) Anti-LAMP1 (L1418) antibody (2 citations)

13 protocols using anti lamp1

1

Visualizing NCOA7 Localization in U87MG Cells

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For visualization of NCOA7 localization by confocal microscopy, U87MG CD4+ CXCR4+ cultures were transduced with lentiviral vectors containing Flag-tagged NCOA7 as described above and were subsequently plated on coverslips treated with poly-L-lysin. Fixation, staining with anti-Flag antibody (Sigma-Aldrich F3165 or F7425, or Miltenyi 130-101-576), anti-EEA1 (BD Biosciences 610457), anti-CD63 (Santa-Cruz sc5275) and anti-Lamp1 (Sigma-Aldrich L1418), and Alexa-488 and Alexa-546 conjugated anti-mouse or anti-rabbit secondary, and and image acquisition was performed as in previous sections with either a Zeiss LSM880 confocal microscope.
Doyle T., Moncorgé O., Bonaventure B., Pollpeter D., Lussignol M., Tauziet M., Apolonia L., Catanese M.T., Goujon C, & Malim M.H. (2018). The interferon inducible isoform of NCOA7 inhibits endosome-mediated viral entry. Nature microbiology, 3(12), 1369-1376.
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2

Comprehensive Antibody Collection for Cell Signaling

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Antibodies used in this study included anti-STING (13647; Cell Signaling Technology), anti-Flag M2 (F3165; Sigma-Aldrich), anti-HA (H3663; Sigma-Aldrich), anti-EGFP (GL-8; Clontech), anti-LC3 (7543; Sigma-Aldrich), anti-p62 (PM045; MBL), anti-STX17 (HPA001204; Sigma-Aldrich), anti-SNAP29 antibody (111303, SYSY, or sc-135564; Santa Cruz Biotechnology), anti-LAMP1(L1418; Sigma-Aldrich), anti-LAMP2 (sc-18822; Santa Cruz or L0668; Sigma-Aldrich), anti-Myc (9E10, DSHB), anti-WIPI2 (ab105459; Abcam), anti-FIP200 (17250; ProteinTech), anti-ATG16L (PM040; MBL), anti-α-Tubulin (E7; DSHB), anti-VAMP8 (ab76021; Abcam), anti-GABARAP (ab109364; Abcam), anti-AMPK (2532; Cell Signaling Technology), anti-p-AMPK T172 (2535; Cell Signaling Technology), anti-TBC1D1(66433; Cell Signaling Technology), anti-p-TBC1D1 Ser237(07-2268; Sigma-Aldrich), anti-Raptor (2280; Cell Signaling Technology), anti-p-Raptor Ser792 (2083; Cell Signaling Technology), anti-TSC2 (4308; Cell Signaling Technology), anti-p-TSC2 Ser1387 (5584; Cell Signaling Technology), anti-ACC (3662; Cell Signaling Technology), anti-p-ACC Ser79 (3661; Cell Signaling Technology), anti-Glut4 (GT-41-A; Alpha diagnostic international), anti-Laminin2 (L0663; Sigma-Aldrich), anti-CD36 (80080; Abcam), anti-Dystrophin (15277; Abcam), and LysoTracker (ENZ-51005; Enzo).
Rong Y., Zhang S., Nandi N., Wu Z., Li L., Liu Y., Wei Y., Zhao Y., Yuan W., Zhou C., Xiao G., Levine B., Yan N., Mou S., Deng L., Tang Z., Liu X., Kramer H, & Zhong Q. (2022). STING controls energy stress-induced autophagy and energy metabolism via STX17. The Journal of Cell Biology, 221(7), e202202060.
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3

Protein Expression Analysis in Cell Lysates

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Cells were lysed with RIPA buffer (Pierce) plus 10 mM NaF, 1 mM Na3VO4 and Protease Inhibitor Cocktail (Sigma). After centrifugation (14,000 x g for 10 min at 4°C), supernatants were collected and protein concentrations were determined with a DC protein assay (Bio-Rad). Proteins (10 μg) were separated by SDS-PAGE (12% gel) and following transfer to polyvinylidene difluoride membranes (Millipore), membranes were incubated overnight at 4°C with antibodies including anti-tau-5 (Abcam), anti-phospho tau (AT8, Thermo Scientific), anti-cathepsin D (Abcam), anti-LAMP-1 (Sigma), and anti-vacuolar-ATPase (Santa Cruz). GAPDH (Abcam) was used as a loading control. The immunoblots were developed with enhanced chemiluminescence, and bands were visualized and analyzed by LabWorks 4.5 software on a UVP Bioimaging System (Upland). Quantification of results was performed by densitometry and the results were analyzed as total integrated densitometric volume values (arbitrary units).
Soliman M.L., Geiger J.D, & Chen X. (2016). Caffeine blocks HIV-1 Tat-induced amyloid beta production and tau phosphorylation. Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, 12(1), 163-170.
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4

Visualizing NCOA7 Localization in U87MG Cells

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For visualization of NCOA7 localization by confocal microscopy, U87MG CD4+ CXCR4+ cultures were transduced with lentiviral vectors containing Flag-tagged NCOA7 as described above and were subsequently plated on coverslips treated with poly-L-lysin. Fixation, staining with anti-Flag antibody (Sigma-Aldrich F3165 or F7425, or Miltenyi 130-101-576), anti-EEA1 (BD Biosciences 610457), anti-CD63 (Santa-Cruz sc5275) and anti-Lamp1 (Sigma-Aldrich L1418), and Alexa-488 and Alexa-546 conjugated anti-mouse or anti-rabbit secondary, and and image acquisition was performed as in previous sections with either a Zeiss LSM880 confocal microscope.
Doyle T., Moncorgé O., Bonaventure B., Pollpeter D., Lussignol M., Tauziet M., Apolonia L., Catanese M.T., Goujon C, & Malim M.H. (2018). The interferon inducible isoform of NCOA7 inhibits endosome-mediated viral entry. Nature microbiology, 3(12), 1369-1376.
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5

Subcellular Fractionation and Immunoblotting

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The cells were washed twice in 1× PBS, pH 7.4, and lysed in buffer A (10 mm HEPES, pH 7.9, 10 mm KCl, 1.5 mm MgCl2, and 0.5% Nonidet P-40). Lysates were spun at 21 kg for 5 min to rid of nuclei. Mitochondria and ER were lysed directly in 1× SDS loading buffer. Solubilized mitochondrial fraction samples were prepared by adding 2× or 4× loading buffer depending on the concentration and the buffers used for solubilization. Protein lysates (50 μg) were resolved by SDS-PAGE, transferred to nitrocellulose membranes, and incubated for 1 h with 5% milk TBS-T and overnight with primary antibodies in 5% BSA at 4 °C or for 1–2 h at room temperature. Antibodies included anti-LAMP1 (1:1000) (Sigma–Aldrich, L1418), anti-calnexin (1:2000) (Cell Signaling Technology, 2433S), anti-tubulin (1:10000) (Cell Signaling Technology, 2146), anti-VDAC1 (1:2000) (Abclonal, A0810), anti-His monoclonal (1:1000) (Abclonal, AE003; Abgent, AM1010a), anti-Creb (1:1000) (Ruiying Biological, RLM3428), anti-RNASET2 (1:1000) (Abgent, AP10764a), anti-mortalin (1:10000) (Sigma–Aldrich, G4045), anti-DDP2 (1:1000), and anti-PNPASE (1:5000) (general gifts from Dr. Carla Koehler at UCLA).
Huang J., Liu P, & Wang G. (2018). Regulation of mitochondrion-associated cytosolic ribosomes by mammalian mitochondrial ribonuclease T2 (RNASET2). The Journal of Biological Chemistry, 293(51), 19633-19644.
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6

Comprehensive Immunofluorescence Staining Panel

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Anti‐GFP and anti‐Ki67 (SP6) from Thermo Fisher Scientific; isolectin‐B4, anti‐MYO1C, and anti‐α‐tubulin (B‐5‐1‐2), anti‐LAMP1, anti‐PCNA (PC10) from Sigma‐Aldrich; anti‐CD31, anti‐FLK1‐APC‐conjugated (Avas12alpha1), anti‐CD71‐FITC‐conjugated (C2), anti‐CD117 (2B8)‐PE‐cy7‐conjugated, anti‐CD31‐FITC‐conjugated (Mec 13.3), anti‐CD102, anti‐IgG2a k isotype‐APC‐conjugated (R3595), anti‐IgG2bk isotype‐PE‐Cy7 conjugated (2B8), anti‐CD326, anti‐Rab4, and anti‐Rab5 from BD Biosciences; anti‐VEGFR2 (55B11), anti‐p‐Tyr‐1175‐VEGFR2 (D5B11), anti‐PLCγ‐1, anti‐p‐Tyr‐783‐PLCγ‐1, anti‐ERK1/2, anti‐p‐ERK‐1/2 (T202/Y204; E10), anti‐Src (36D10), anti‐p‐Src (Tyr416; D49G4), anti‐CDK4 (D9G3E), anti‐Rb (4H1), anti‐p‐Rb (Ser780; C84F6), and anti‐p‐Rb (Ser807/811; D20B12), anti‐PCNA (PC10), anti‐ULK1 (D8H5), anti‐ATG9A (D409D) from Cell Signaling Technology; anti‐endomucin (V.7C7), anti‐Flk‐1 (A‐3), anti‐podocin (G‐20), anti‐caveolin‐1 (N‐20) and anti‐E2F2 (TFE‐25), anti‐E2F1 from Santa Cruz Biotechnology; anti‐human VEGFR2 (89109) from R&D System; anti‐TFEB from MyBiosource; hypoxyprobe‐1‐FITC‐conjugated antibody (Chemicon); anti‐LC3 from Novus Biologicals; anti‐cyclin D1 (SP4), anti‐E2F1, and anti‐TGN46 (2F7.1) from Abcam.
Doronzo G., Astanina E., Corà D., Chiabotto G., Comunanza V., Noghero A., Neri F., Puliafito A., Primo L., Spampanato C., Settembre C., Ballabio A., Camussi G., Oliviero S, & Bussolino F. (2018). TFEB controls vascular development by regulating the proliferation of endothelial cells. The EMBO Journal, 38(3), e98250.
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7

Phagosome Protein Profiling by Western Blot

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Visceral organs including the lungs, liver, and kidneys harvested at various time periods were homogenized, lysed, and centrifuged to collect the supernatant containing cytoplasmic proteins. Isolated BMMs were treated with CM or taurolidine for 12 h and allowed to phagocytose E. coli for 1 h. Phagosomes were purified using a sucrose gradient as described previously (52 (link)), and phagosomal proteins were extracted. Equal amounts of protein extracts were separated on sodium dodecyl-polyacrylamide gels and transblotted onto nitrocellulose membranes (Schleicher & Schuell). The membrane was blocked for 1 h with 0.05% Tween-20 containing 5% nonfat milk and probed overnight at 4 °C with anti-LC3B (Abcam), anti-LAMP-1 (Sigma-Aldrich), anti-Rubicon (Cell Signaling), anti-β-tubulin (Sigma-Aldrich), anti-PDI (Cell Signaling), and anti-UNC93B (Abcam) antibodies. Blots were then incubated with horseradish-peroxidase-conjugated secondary antibodies, developed with SuperSignal chemiluminescent substrate (Pierce), and captured with an LAS-3000 imaging system (Fujifilm).
Huang J., Ita M., Zhou H., Zhao H., Hassan F., Bai Z., O’Leary D.P., Li Y., Redmond H.P., Wang J.H, & Wang J. (2022). Autophagy induced by taurolidine protects against polymicrobial sepsis by promoting both host resistance and disease tolerance. Proceedings of the National Academy of Sciences of the United States of America, 119(19), e2121244119.
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8

Immunoblotting and Immunocytochemistry Protocols

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The followings are primary antibodies used: mouse monoclonal anti-SQSTM1/p62 (Abcam, ab56416; 1:20,000 for WB, 1:300 for ICC), rabbit polyclonal anti-LC3 (Sigma-Aldrich, L7543; 1:10,000 for WB, 1:300 for ICC), rabbit polyclonal anti-LAMP1 (Sigma-Aldrich, L1418; 1:200 for ICC), rabbit polyclonal anti-GAPDH (BioWorld, AP0066; 1:5,000 for WB), rabbit polyclonal anti-β-actin (BioWorld, AP0060; 1:5,000 for WB), mouse monoclonal anti-α-synuclein (BD Transduction, 610,787; 1:2000 for WB, 1:300 for ICC), rabbit monoclonal anti-α-synuclein (phospho-S129) (Abcam, ab51253; 1:200 for ICC, 1:500 for IHC), mouse monoclonal anti-c-Myc (Santa Cruz Biotechnology, H1721; 1:2,000 for WB) rabbit polyclonal anti-DFCP1/ZFYVE1 (ABclonal, A7527; 1:150 for ICC), rabbit monoclonal anti-PARP (Cell Signaling Technology, 9532; 1:1,000 for WB), rabbit monoclonal anti-GFAP (Cell Signaling Technology, D1F4Q; 1:500 for IHC), and mouse monoclonal anti-phospho-Histone H2A.X (ser139) (Upstate, 05–636; 1:200 for ICC). The followings are secondary antibodies used: anti-rabbit IgG-HRP (Cell Signaling Technology, 7074; 1:5,000), anti-mouse IgG-HRP (Cell Signaling Technology, 7076; 1: 5,000), Alexa fluor 488 goat anti-rabbit IgG (A11008; 1:500), and Alexa fluor 555 goat anti-mouse IgG (Invitrogen, A11029; 1:500).
Lee J., Sung K.W., Bae E.J., Yoon D., Kim D., Lee J.S., Park D.H., Park D.Y., Mun S.R., Kwon S.C., Kim H.Y., Min J.O., Lee S.J., Suh Y.H, & Kwon Y.T. (2023). Targeted degradation of ⍺-synuclein aggregates in Parkinson’s disease using the AUTOTAC technology. Molecular Neurodegeneration, 18, 41.
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9

Immunofluorescence Imaging of Endosomal Markers

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Cells were grown to 70% confluence on glass coverslips, fixed for 20 minutes at room temperature in 4% paraformaldehyde, and blocked in 50mM ammonium chloride for 10 minutes and subsequently 0.1% bovine serum albumin in phosphate-buffered saline (PBS) for 20 minutes. Cells were incubated with primary antibodies anti-BMP/LBPA antibody clone 6C4 (MABT837, Millipore Sigma; 1/100), anti-LAMP1 (L1418, Sigma-Aldrich; 1:100) and anti-Rab7 D95F2 (9367, Cell Signaling; 1/100) in PBS containing 0.5% saponin for 30 minutes at room temperature. Following incubation with Alexa Fluor secondary antibodies (1:1000) for 30 minutes, cells were mounted in 4′,6-diamidino-2-phenylindole (DAPI) Fluoromount-G (0100–20, Southern Biotech). Samples were imaged with a Zeiss LSM 710 AxioObserver confocal microscope and analyzed with ZenBlue software.
Butler D.L., Jakielaszek C.J., Miller L.A, & Poupard J.A. (1999). Escherichia coli ATCC 35218 as a Quality Control Isolate for Susceptibility Testing of Haemophilus influenzae with Haemophilus Test Medium. Antimicrobial Agents and Chemotherapy, 43(2), 283-286.
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10

Immunoblotting and Immunocytochemistry Protocols

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The followings are primary antibodies used: mouse monoclonal anti-SQSTM1/p62 (Abcam, ab56416; 1:20,000 for WB, 1:300 for ICC), rabbit polyclonal anti-LC3 (Sigma-Aldrich, L7543; 1:10,000 for WB, 1:300 for ICC), rabbit polyclonal anti-LAMP1 (Sigma-Aldrich, L1418; 1:200 for ICC), rabbit polyclonal anti-GAPDH (BioWorld, AP0066; 1:5,000 for WB), rabbit polyclonal anti-β-actin (BioWorld, AP0060; 1:5,000 for WB), mouse monoclonal anti-α-synuclein (BD Transduction, 610,787; 1:2000 for WB, 1:300 for ICC), rabbit monoclonal anti-α-synuclein (phospho-S129) (Abcam, ab51253; 1:200 for ICC, 1:500 for IHC), mouse monoclonal anti-c-Myc (Santa Cruz Biotechnology, H1721; 1:2,000 for WB) rabbit polyclonal anti-DFCP1/ZFYVE1 (ABclonal, A7527; 1:150 for ICC), rabbit monoclonal anti-PARP (Cell Signaling Technology, 9532; 1:1,000 for WB), rabbit monoclonal anti-GFAP (Cell Signaling Technology, D1F4Q; 1:500 for IHC), and mouse monoclonal anti-phospho-Histone H2A.X (ser139) (Upstate, 05–636; 1:200 for ICC). The followings are secondary antibodies used: anti-rabbit IgG-HRP (Cell Signaling Technology, 7074; 1:5,000), anti-mouse IgG-HRP (Cell Signaling Technology, 7076; 1: 5,000), Alexa fluor 488 goat anti-rabbit IgG (A11008; 1:500), and Alexa fluor 555 goat anti-mouse IgG (Invitrogen, A11029; 1:500).
Lee J., Sung K.W., Bae E.J., Yoon D., Kim D., Lee J.S., Park D.H., Park D.Y., Mun S.R., Kwon S.C., Kim H.Y., Min J.O., Lee S.J., Suh Y.H, & Kwon Y.T. (2023). Targeted degradation of ⍺-synuclein aggregates in Parkinson’s disease using the AUTOTAC technology. Molecular Neurodegeneration, 18, 41.
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