Lamp1 h4a3

Manufactured by Developmental Studies Hybridoma Bank

Sourced in United States

LAMP1 (H4A3) is a monoclonal antibody produced by the Developmental Studies Hybridoma Bank. It is designed to detect the lysosome-associated membrane protein 1 (LAMP1), which is a widely expressed integral membrane protein found in the lysosomal membrane. The antibody can be used for various applications, such as immunostaining and Western blotting, to study the localization and expression of LAMP1 in cells and tissues.

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Lab products found in correlation

Dapi 4 6 diamidino 2 phenylindole, by Thermo Fisher Scientific (1 mentions) Atg5 d5f5u, by Cell Signaling Technology (1 mentions) Z vad fmk, by Apexbio (1 mentions) Horseradish peroxidase conjugated anti rabbit and anti mouse, by GE Healthcare (1 mentions) Amino acids, by Thermo Fisher Scientific (1 mentions) P s6k1 t389, by Cell Signaling Technology (1 mentions) Histone h4, by Cell Signaling Technology (1 mentions) Calnexin, by Abcam (1 mentions) Z vad fmk s7023, by Selleck Chemicals (1 mentions) Lc3b l7543, by Merck Group (1 mentions) Atg4b 15 131 1 ap, by Proteintech (1 mentions) Gapdh sc 365062, by Santa Cruz Biotechnology (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Opti mem, by Thermo Fisher Scientific (1 mentions) Gapdh fl 335, by Santa Cruz Biotechnology (1 mentions) Flag f7425, by Merck Group (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Dnase 2, by Merck Group (1 mentions)

Spelling variants of this product

LAMP1 (28 citations) Mouse anti-LAMP1 (H4A3, (4 citations) LAMP1 antibody (H4A3), (3 citations) Anti-LAMP1 (H4A3) (3 citations) LAMP1 clone H4A3 (2 citations)

6 protocols using lamp1 h4a3

Regulation of Autophagy and Apoptosis

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Chloroquine (CQ), staurosporine (STP), E64 and cisplatin (cis-diaminedichloroplatinum (II), cisPt) were obtained from Sigma-Aldrich (St. Louis, MO). Dextran fluorescein 40,000 MW and DAPI (4’, 6-diamidino-2-phenylindole) were purchased from Life Technologies (Grand Island, NY). The pan-caspase inhibitor z-VAD-FMK was purchased from ApexBio Technology (Houston, TX). The primary antibodies used in this study includes the following: caspase 3 (3G2), LC3 A/B, ATG5 (D5F5U) and p53 (1C12) from Cell Signaling Technology (Beverly, MA), cathepsin D (H-75) and Bax (N-20) from Santa Cruz Biotechnology (Santa Cruz, CA), LAMP-1 (H4A3) from Developmental Studies Hybridoma Bank at the University of Iowa, BAK from EMD Millipore (Billerica, MA), and α-Tubulin Ab2 (clone DM1A) from Fisher Scientific (Rockford, IL). The secondary antibodies, horseradish peroxidase conjugated anti-rabbit and anti-mouse, were both from GE Healthcare (Pittsburgh, PA). ECL2 was purchased from Thermo Fisher Scientific (Rockford, IL).

Circu M., Cardelli J., Barr M., O’Byrne K., Mills G, & El-Osta H. (2017). Modulating lysosomal function through lysosome membrane permeabilization or autophagy suppression restores sensitivity to cisplatin in refractory non-small-cell lung cancer cells. PLoS ONE, 12(9), e0184922.

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Antibody Characterization and Validation

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Antibodies: pS6K1 (T389, #9205), S6K1 (#9202), FLCN (rabbit monoclonal, #3697) and histone H4 (#2935) were obtained from Cell Signalling Technology (Danvers, MA); FLCN (mouse monoclonal, #271558) and bafilomycine A1 was from Santa Cruz Biotechnology; LAMP1 (H4A3) and GFP (12A6 and 8H11) were purchased from Developmental Studies Hybridoma Bank; calnexin (#22595) and HA (#9110) was obtained from Abcam; GFP (A11122) and all fluorescent secondary antibodies were purchased from Life Technologies; amino acids were from Sigma Aldrich.

Wu X., Zhao L., Chen Z., Ji X., Qiao X., Jin Y, & Liu W. (2016). FLCN Maintains the Leucine Level in Lysosome to Stimulate mTORC1. PLoS ONE, 11(6), e0157100.

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Autophagy-Related Protein Detection

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Antibodies used in this study were as follow: ATG4B (15131-1-AP) was from Proteintech (Wuhan, China), ATG5 (8540) and FIP200 (12436) were from Cell Signaling Technology (Danvers, MA, USA), GAPDH (sc-365062) was from Santa Cruz Biotechnology (Dallas, TX, USA), LC3B (L7543) and TUBA (T6074) were from Sigma (St. Louis, MO, USA), LAMP1 (H4A3) was from Developmental Studies Hybridoma Bank (Iowa City, IA, USA), SQSTM1/p62 (PM045) was from MBL International (Woburn, MA, USA). Secondary antibodies (35503, 35511, 35553, and 35561) were purchased from Thermo Fisher Scientific (Rockford, IL, USA).
Bafilomycin A1 (Baf, B-1080) was purchased from LC Laboratories (Woburn, MA, USA), as well as rapamycin (Rap, ASW-125). Chloroquine (CQ, A506569) was purchased from Sangon Biotech (Shanghai, China). Z-VAD-FMK (S7023) and staurosporine (SP, S1421) was from Selleckchem (Houston, TX, USA). Dequenched-BSA (DQ-BSA) Red (D12051), LysoTracker Red (LTR, L12492) and Lipofectamine 2000 (11668-019) were purchased from Life Technologies (Bothell, WA, USA). Substrate Ac-DEVD-AFC (CASP-048) for caspase3 activity was from Chinese Peptide Company (Hangzhou, China). Purified ATG4B, substrate ECFP-GATE-16-YFP (FRET-GATE16) for ATG4B were described before [45 (link)].

Fu Y., Gu Q., Luo L., Xu J., Luo Y., Xia F., Han F., Hong L., Yin X.M., Huang Z, & Li M. (2020). New Anti-Cancer Strategy to Suppress Colorectal Cancer Growth Through Inhibition of ATG4B and Lysosome Function. Cancers, 12(6), 1523.

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Molecular and Cellular Characterization of HCV Infection

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General materials were purchased from Sigma-Aldrich or VWR. DMEM, FBS, Opti-MEM, and Lipofectamine 3000 were purchased from Thermo Fisher Scientific. Protease inhibitor cocktail (P8340; Sigma-Aldrich) was used at 1:100 dilution. Monoclonal antibody C7-50 to HCV core protein and monoclonal antibody JL-8 to GFP were purchased from Thermo Fisher Scientific. RILP (H85), ApoE (A1.4), CD63 (MX-49.129.5), CD81 (5A6), and GAPDH (FL-335) were purchased from Santa Cruz Biotechnology. Rab7 (D95F2), histone H2B (2722), β-Tubulin, Flotillin-1 (D2V7J), Argonaute 2 (C34C6), Hrs (D7T5N), NLRP3 (D4D8T), FMRP (4317S), GM130 (D6B1), Tom20 (D8T4N), and Caspase-4 (4450) were purchased from Cell Signaling Technology. FMR1 (F4055) and FLAG (F7425) were purchased from Sigma-Aldrich. Caspase-1 (EPR19675), IL-1β (EPR21086), TSG101 (EPR7130(B)), gasdermin D (EPR19829), and HA (HA.C5) were purchased from Abcam. Lamp1 (H4A3) was purchased from Developmental Studies Hybridoma Bank.

Wozniak A.L., Adams A., King K.E., Dunn W., Christenson L.K., Hung W.T, & Weinman S.A. (2020). The RNA binding protein FMR1 controls selective exosomal miRNA cargo loading during inflammation. The Journal of Cell Biology, 219(10), e201912074.

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Western Blot Analysis of Cell Lysates

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After 5 days of culture in various media, cells were lysed for Western Blot according to our published protocol.[33 (link)] Cell lysates were evaluated by primary antibodies specific to proliferating cell nuclear antigen (PCNA, D3H8P, 1: 1000, Cell Signaling Technology, Danvers, MA), sterol regulatory element-binding protein 1(SREBP-1, C-20, 1:500, Santa Cruz Biotechnology), lysosomal-associated membrane protein 1 (Lamp-1, H4A3, 1:500, Developmental Studies Hybridoma Bank), DNase II (1: 1000, Sigma-Aldrich) or β-actin (1:10,000, Cell Signaling Technology). Secondary antibodies were horseradish peroxidase (HRP)-conjugated goat anti-rabbit or goat anti-mouse IgG (both 1:5000, Sigma-Aldrich Corp.). Densitometry was performed by using ImageJ (http://rsb.info.nih.gov/ii).

Liu Y., Chen D., Chen X., Kam W.R., Hatton M.P, & Sullivan D.A. (2018). Hypoxia: A breath of fresh air for the meibomian gland. The ocular surface, 17(2), 310-317.

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Immunostaining of Human Eyelid Sections

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Human eyelid sections and HMGECs were fixed with cold methanol for 15 minutes at −20°C. Following three phosphate‐buffered saline (PBS) rinses for 5 minutes each, samples were blocked with 2% bovine serum albumin (BSA, Sigma‐Aldrich Corp., St. Louis, MO) in PBS for 60 minutes, and then incubated overnight at 4°C in a moist chamber with antibodies specific for Lrig1 (ab214102,1:100), cytokeratin 14 (ab181595, 1:500), cytokeratin 6 (ab18586, 1:500), DNase2 (ab8119, 1:100; Abcam, Cambridge, MA), or lysosomal‐associated membrane protein 1 (LAMP‐1;H4A3, 1:15; Developmental Studies Hybridoma Bank, Iowa City, IA), or the BSA diluent. After three additional PBS rinses, donkey anti‐rabbit (ab150075, 1:500, Abcam) or donkey anti‐mouse (2492098, 1:500, EMD Millipore, Temecula, CA) secondary antibodies were applied for 1 hour at room temperature. For neutral lipid staining, some eyelid sections were fixed in 4% paraformaldehyde for 15 minutes. Following additional washes, samples were exposed to LipidTOX Green neutral lipid stain (1:500, Thermo Fisher Scientific) in a humid chamber for 30 minutes. Nuclei were counterstained with 4′,6‐diamidino‐2‐phenylindole (DAPI, 1 μg/mL, Sigma‐Aldrich) and samples were covered with VectaMount mounting medium (Vector Laboratories, Burlingame, CA), and observed with a confocal microscope (Leica TCS SP8, Leica Microsystems, Wetzlar, Germany).

Xie H., Sullivan D.A., Chen D., Hatton M.P., Kam W.R, & Liu Y. (2018). Biomarkers for Progenitor and Differentiated Epithelial Cells in the Human Meibomian Gland. Stem Cells Translational Medicine, 7(12), 887-892.

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