N-succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin (suc-LLVY-MCA) was from Sigma-Aldrich (St Louis, MO). The antibodies used in this study were anti-CerS2 (Sigma Aldrich, St Louis, MO), anti-cathepsin D, anti-cathepsin L and anti-cathepsin S (Santa Cruz Biotechnology, Santa Cruz, CA), anti-HNE-Michael adducts (Calbiochem, San Diego, CA), anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Millipore, Temecula, CA) and anti-F4/80 (Biolegend, San Diego, CA).
Anti cathepsin d
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-cathepsin D is a laboratory reagent used to detect and quantify the presence of cathepsin D, a lysosomal aspartic protease, in biological samples. It can be used in various research applications, such as cell biology, molecular biology, and biochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Santa Cruz Biotechnology (3 mentions)
3 methyladenine, by Merck Group (2 mentions)
Anti lc3, by Abcam (2 mentions)
Anti bax, by Santa Cruz Biotechnology (2 mentions)
Anti lamp1, by Santa Cruz Biotechnology (2 mentions)
Anti phospho mek1 2, by Cell Signaling Technology (2 mentions)
Anti phospho iκbα, by Cell Signaling Technology (2 mentions)
Anti iκbα, by Santa Cruz Biotechnology (2 mentions)
Anti phospho erk1 2, by Cell Signaling Technology (2 mentions)
Anti lc3b, by Cell Signaling Technology (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Anti cathepsin l, by Santa Cruz Biotechnology (1 mentions)
Anti cers2, by Merck Group (1 mentions)
Anti cathepsin s, by Santa Cruz Biotechnology (1 mentions)
Anti glyceraldehyde 3 phosphate dehydrogenase gapdh, by Merck Group (1 mentions)
Anti f4 80, by BioLegend (1 mentions)
Anti mtor, by Santa Cruz Biotechnology (1 mentions)
Pro q diamond, by Thermo Fisher Scientific (1 mentions)
Anti asc antibody, by Santa Cruz Biotechnology (1 mentions)
Sypro ruby, by Thermo Fisher Scientific (1 mentions)
17 protocols using anti cathepsin d
1
Cathepsin and Lipid Peroxidation Assay
2
Synthesis and Characterization of Modified Carbon Nanotubes and Nanoparticles
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AP-MWCNTs were purchased from Cheap Tubes. COOH-MWCNTs were synthesized by oxidizing AP-MWCNTs in mixed acids.4 (link) Min-U-Sil silica (quartz) was purchased from US Silica (Frederick, MD, USA). Mesoporous silica was generously provided by Dr. Jeffrey Zink, Department of Chemistry and Biochemistry at UCLA. La2O3, Gd2O3, Sm2O3, Yb2O3 nanoparticles were purchased from Nanostructured & Amorphous Materials (Houston, TX, USA). TiO2 and Bi2O3 were provided by Dr. Lutz Madler at the Department of Production Engineering, University of Bremen, Germany. CQ, Sypro Ruby, Pro-Q-Diamond, LAMP-1 antibody and pHrodo were purchased from Life Technologies (Grand Island, NY, USA). 4-Methylumbelliferyl-beta-d -galactopyranoside, 3-methyladenine and Rapa were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-LC-3 was purchased from Abcam (Cambridge, MA, USA). Anti-mTOR, anticathepsin D and anti-ASC antibodies were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). Recombinant human β-galactosidase was purchased from R&D (Minneapolis, MN, USA). The phosphopeptide, LPSSPVpYEDAASFK, was purchased from Apeptide (Shanghai, China). NLRP3–/– or ASC–/– THP-1 cells are prepared from THP-1 cells that transfected with NLRP3 or ASC shRNA.
3
Western Blot Analysis of SOD1 in DRG
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For Western blot (WB) analysis, DRG were homogenized in a lysis buffer (0.1 mol/L NaCl, 0.01 mol/L Tris-HCl, pH 7.5, 1 mmol/L EDTA, and 1 μg 7 Ml aprotinin) and the homogenates were centrifuged. Overall, 3 SOD1 and 3 control mice were used, respectively. The protein in the supernatants were subjected to SDS-PAGE electrophoresis and afterwards transferred to polyvinylidene difluoride membranes, stained with antibodies and visualized with the Odyssey system (LI-COR Biotechnology). Alpha-Tubulin was used as protein loading control. The antibodies used were rabbit polyclonal anti-SOD1 (dilution 1:1,000; Enzo, Life Sciences, Switzerland), goat polyclonal antibody anti-cathepsin D (dilution 1:2,000; Santa Cruz Biotechnology, USA) and mouse monoclonal anti-Tubulin (dilution 1:000; Abcam, Cambridge, USA). ImageJ software (U.S. National Institutes of Health, Bethesda, Maryland, USA) was used to quantify the density and size of the blots.
4
Macrophage Response to M. tuberculosis
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Macrophages were infected with M. tuberculosis until the indicated time points. Western blot assays were performed as previously described (Lee et al., 2015 (link)). For the Western blot assays, anti-iNOS, anti-cathepsin D, and anti-actin antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The housekeeping protein, actin, was used to confirm that the well were equal loaded.
5
Western Blot Analysis of CLCN3 and Apoptosis Markers
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Cells were harvested and lysed in RIPA lysis buffer (Beyotime, Shanghai, China) that included a protease inhibitor cocktail (19 (link)) (Sigma-Aldrich; Merck KGaA). Protein quantification was performed using the Bio-Rad Protein assay dye reagent (Bio-Rad Laboratories, Inc., Hercules, CA, USA). Proteins were electrophoresed and separated by SDS-PAGE, and then transferred from gels onto PVDF membranes (EMD Millipore, Billerica, MA, USA). After incubation with primary (4°C, overnight) and secondary antibodies (room temperature, 2 h), the blots were developed with DAB (3,3′-diaminobenzidine). Western blots results were analyzed with GIS 1D gel image system software. The following primary antibodies were used: anti-CLCN3 (1:100, sc-17572), anti-Bcl-2 (1:200, sc-783), anti-Bax (1:200, sc-7480), anti-pro-caspase 3 (1:200, sc-7148), anti-cleaved-caspase 3 (1:200, sc-22171), anti-cathepsin D (1:200, sc-136282), anti-β-actin (1:400, sc-47778; Santa Cruz Biotechnology, Inc., Dallas, TX, USA). Secondary HRP-conjugated antibodies were purchased from Thermo Fisher Scientific, Inc. Densitometry was used to calculate the ratio of CLCN3 to β-actin, cleaved caspase 3 to pro-caspase 3, and BCL2 to BAX signal in each lane.
6
Cell Culture and Reagent Preparation
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Cells were obtained from the American Type Culture Collection (Manassas, VA, USA). All cells were cultured in appropriate medium containing 10% fetal bovine serum (FBS; Welgene, Gyeongsan, Korea), 1% penicillin-streptomycin, and 100 μg/mL gentamycin (Thermo Fisher Scientific, Waltham, MA, USA). All cell lines tested negative for mycoplasma contamination. The lines were authenticated by standard morphologic examination using microscopy. FTY720 was purchased from Echelon Biosciences (Salt Lake City, UT, USA). N-acetyl-l -cysteine was obtained from Calbiochem (San Diego, CA, USA). E64D and NBD-FTY720 were purchased from Cayman Chemical (Ann Arbor, MI, USA). TNF-α and z-VAD-fmk were purchased from R&D Systems (Minneapolis, MN, USA). PD98059, SB203580, SP600125, NecroX-5, and pepstatin A were obtained from Enzo Life Science (Farmington, NY, USA). Anti-PARP, anti-Cathepsin D, anti-LAMP1, and anti-Beclin 1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-phospho-ERK, phosho-p38, anti-phospho-JNK, and anti-Alix were purchased from Cell Signaling Technology (Beverly, MA, USA). The anti-ATG7 antibody was obtained from ProSci Inc. (Poway, CA, USA). The cycloheximide, H2O2, 3-methyladenine, actinomycin D, glutathione ethyl ester, and anti-actin antibody were obtained from Sigma (St. Louis, MO, USA).
7
Mtb Infection-Induced Signaling Pathways
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Macrophages were infected with Mtb for the indicated times. Cells were lysed with lysis buffer consisting of 10 mM Tris–HCl, 1 mM EDTA, 140 mM NaCl, 0.1% DOC, 0.1% SDS, and Triton X-100 containing complete protease inhibitor cocktail (Calbiochem, San Diego, CA, USA). Western blotting was performed as previously described (27 (link)). Anti-phospho-GSK3β, anti-IκBα, anti-cathepsin D, and anti-β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-phospho-PI3K, anti-PI3K, anti-phospho-p38, anti-p38, anti-phospho-ERK1/2, anti-phospho-MEK1/2, anti-phospho-JNK1/2, anti-phospho-IκBα, and anti-IRAK-M were purchased from Cell Signaling Technology.
8
Cell Death Mechanisms Evaluation
9
Antibody Validation for Peroxisomal Proteins
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Anti-PEX5 was purchased from GeneTex (Irvine, CA, USA). Anti-catalase, anti-LAMP-1, anti-PCNA, anti-cathepsin B, and anti-cathepsin D were bought from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-LAMP-2 and anti-LC3 were obtained from Abcam (Cambridge, MA, USA). Anti-TFEB was purchased from MyBioSource (San Diego, CA, USA). Anti-TFEB, anti-p-p70S6K, anti-p70S6K, anti-p-S6R, anti-S6R, anti-p-4E-BP-1, anti-4E-BP-1, and anti-TSC2 were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-PMP70 was bought from Thermo Fisher Scientific (Waltham, MA, USA). Anti-ACOX1 was purchased from Proteintech (Chicago, IL, USA). Anti-DBP (HSD17B4) was bought from OriGene Technologies (Rockville, MD, USA). Anti- SQSTM1/p62 was obtained from Sigma-Aldrich (St. Louis, MO, USA).
10
Antibody Sourcing for Cellular Protein Analysis
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Anticathepsin D, anti–c-Jun, and anti–Egr-1 were from Santa Cruz; anti–Iba-1 was from Wako; anti-CD44 was from Cell Signaling; antivimentin was from Dako; anti-GAPDH, antidesmoplakin, anti– HMOX-1, anti-Prdx6, anti- ALDH1A1, and anti- IGF1 were from Abcam; antiheat-shock protein 27 (hsp27) was from Novus; anticlusterin was from Chemicon, anticystatin C was from Upstate; and anti–α-actin was from Sigma.
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