Anti lamp1

Manufactured by Santa Cruz Biotechnology

Sourced in United States, Germany

Anti-LAMP1 is a primary antibody that targets the lysosomal-associated membrane protein 1 (LAMP1). LAMP1 is a glycoprotein that is primarily localized in the membrane of lysosomes and late endosomes. This antibody can be used to detect and study the expression and distribution of LAMP1 in various cell and tissue samples.

Automatically generated - may contain errors

Lab products found in correlation

Anti p 4ebp1, by Cell Signaling Technology (4 mentions) Axiovert 200m, by Zeiss (4 mentions) Anti mtor, by Cell Signaling Technology (3 mentions) Anti 4e bp1, by Cell Signaling Technology (3 mentions) Anti cathepsin b, by Santa Cruz Biotechnology (3 mentions) Anti flag, by Merck Group (3 mentions) Anti cox 4, by Cell Signaling Technology (2 mentions) Anti β actin, by Santa Cruz Biotechnology (2 mentions) Anti tom20, by Santa Cruz Biotechnology (2 mentions) Anti ubiquitin, by Santa Cruz Biotechnology (2 mentions) Anti eea1, by Cell Signaling Technology (2 mentions) Anti cathepsin d, by Santa Cruz Biotechnology (2 mentions) Anti parp, by Santa Cruz Biotechnology (2 mentions) 3 methyladenine, by Merck Group (2 mentions) Anti lamin a c, by Cell Signaling Technology (2 mentions) Anti gapdh, by Santa Cruz Biotechnology (2 mentions) Anti hsp60, by Santa Cruz Biotechnology (2 mentions) Chloroquine, by Merck Group (2 mentions) Anti ha, by Santa Cruz Biotechnology (2 mentions) Anti rab7, by Cell Signaling Technology (2 mentions)

Spelling variants of this product

LAMP1 (83 citations) Mouse anti-LAMP1 (14 citations) Anti-LAMP1 (H4A3) (7 citations) Rat anti-mouse LAMP1 (3 citations) Mouse monoclonal anti-LAMP1 (3 citations) Mouse monoclonal anti-LAMP1 (H4A3) (3 citations) Goat anti‐LAMP1 (2 citations) Anti-Lamp1 (clone H4A3, (2 citations) Rat anti-LAMP1 (1D4B) (2 citations) Mouse anti-Lamp1 (H4A3, (2 citations)

46 protocols using anti lamp1

Antibody Staining and Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

3-(4,5-dimethylthiazol-diphenyltetrazolium bromide (MTT), was purchased from Sigma Aldrich (Milan, Italy). The following rabbit polyclonal antibodies (Abs) were used: Anti-histone H3 (1:1000; Cell Signaling Technology, Danvers, MA, USA), anti-COX IV (1:250; Cell Signaling Technology) and anti-caveolin-1 (1:400; Cell Signaling Technology). The following mouse monoclonal Abs were used: anti-TRPML1 (clone F-10, 1:300 for Western blot, 1:50 for FACS analysis; Santa Cruz Biotechnology, Dallas, TX, USA), anti-TRPML2 (1:300 for Western blot; Santa Cruz Biotechnology), anti-LAMP1 (1:300; Santa Cruz Biotechnology), anti-calreticulin (1:1000; BD Biosciences, Milan, Italy) and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH, 14C10, 1:1000; Cell Signaling Technology). anti-TRPML2 from Sigma Aldrich was used diluted 1:50 for FACS analysis.
The following secondary antibodies were used: horseradish peroxidase (HRP)-conjugated anti-mouse IgG and HRP-conjugated anti-rabbit IgG (1:2000; Cell Signaling Technology), FITC-conjugated anti-rabbit Ab (BD Biosciences), PE-conjugated anti-mouse Ab (BD Biosciences, Milan, Italy), Alexa Fluor-594-conjugated anti-mouse Ab (1:100; Invitrogen, San Diego, CA, USA), Alexa Fluor-488-conjugated anti-mouse Ab (1:100; Invitrogen) and Alexa Fluor-488-conjugated anti-rabbit Ab (1:100; Invitrogen).

Santoni G., Maggi F., Amantini C., Arcella A., Marinelli O., Nabissi M., Santoni M, & Morelli M.B. (2022). Coexpression of TRPML1 and TRPML2 Mucolipin Channels Affects the Survival of Glioblastoma Patients. International Journal of Molecular Sciences, 23(14), 7741.

+ Open protocol

+ Expand

Intracellular Trafficking Regulation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Most general reagents used in this study were sourced from Sigma-Aldrich. The siRNA oligonucleotides were purchased from Dharmacon. Primary antibodies used in this study were as follows: anti-TBC1D5 [Santa Cruz, sc-376296, dilution 1:400 or 1:1000 for immunofluorescence (IF) microscopy or western blotting (WB), respectively], anti-VPS26 (Abcam, ab23892, 1:800 IF or 1:1000 WB), anti-VPS35 [Santa Cruz, sc-374372, 1:800 IF or 1:1000 WB, or from the Seaman lab (see Seaman, 2007 (link)), 1:300 for IF], anti-CIMPR (Abcam, ab2733, 1:400 IF or 1:1000 WB), anti-Lamp1 (Santa Cruz, sc-18821, 1:500 IF or 1:1000 WB), anti-Glut1 (Abcam, ab15309, 1:400 IF), anti-GM130 (BD Transduction labs 610822, 1:500 IF), anti-Fam21 (Millipore, ABT79, 1:400 IF or 1:1000 WB), anti-Aβ (Covance, SIG-39320, 1:1000 WB), anti-sAPPβ (IBL America, 10321, 1:800 WB), anti-Rab7a:GTP (NewEast Biosciences, 26923, 1:300 IF), anti-TGN46 (Seaman lab, see Seaman, 2007 (link), dilution 1:600 IF), anti-GFP (Seaman lab, see Seaman et al., 2009 (link), 1:1000 for immunoprecipitation), anti-Snx1 (BD Transduction labs, dilution 1:400 IF or 1:1000 WB) and anti-Tubulin (Sigma-Aldrich, dilution 1:1000 WB). Secondary fluorescently labelled antibodies were purchased from Invitrogen.

Seaman M.N., Mukadam A.S, & Breusegem S.Y. (2018). Inhibition of TBC1D5 activates Rab7a and can enhance the function of the retromer cargo-selective complex. Journal of Cell Science, 131(12), jcs217398.

+ Open protocol

+ Expand

Western Blot Analysis of Cell Signaling

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot analysis was performed as described previously28 (link), using equal amounts (10 or 50 μg) of protein from each cell lysate. The following antibodies were used: anti-SerpinB2, anti-uPA, anti-LAMP1, anti-phosphorylated p38 (p-p38), anti-p38, anti-phosphorylated ERK (p-ERK), anti-ERK, anti-β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-MMP-2 (Cell Signaling Technology, Danvers, MA, USA); and anti-E-cadherin and anti-N-cadherin (BD Biosciences, San Diego, CA, USA).

Bae S.Y., Park H.J., Hong J.Y., Lee H.J, & Lee S.K. (2016). Down-regulation of SerpinB2 is associated with gefitinib resistance in non-small cell lung cancer and enhances invadopodia-like structure protrusions. Scientific Reports, 6, 32258.

+ Open protocol

+ Expand

Immunofluorescence Analysis of Drug Effects on MCF7 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

MCF7 cells at the concentration of 50,000 cells/well were seeded onto glass coverslips and treated with 5 μmol/L fluoxetine, ebastine, penfluridol, pimozide, fluspirilene, spiperone, nefazodone for 16 h. After the treatment, cells were washed with PBS and fixed with PFA 4% for 10 min at room temperature and washed with PBS. Then cells were permeabilized incubating with cold HEPES-Triton X-100 (20 mM HEPES pH 7.4, 300 mM sucrose, 50 mM NaCl, 3 mM MgCl₂, 0.5% Triton X-100) for 5 min at 4°C. Cells were washed with 0.2% PBS-BSA and saturated using 2% PBS-BSA for 15 min before placing primary antibodies.
Antibodies used in these experiments were anti-mTOR (Cell Signaling Technology), anti-Galectin-1 (Santa Cruz Biotechnology), anti-LAMP1 (Santa Cruz Biotechnology). Cells were incubated with primary antibodies for 30 min, then washed, saturated with 2% PBS-BSA and incubated with secondary antibodies conjugated with Alexa Fluor-488, −536 (Invitrogen) and DAPI for 30 min.
After the incubation, glasses were mounted on glass slides using Mowiol (20% Mowiol 4–88, 2.5% DABCO in PBS, pH 7.4). Images were acquired at confocal microscope Leica TCS SP8 or fluorescence microscope DM5500B (Leica) and analyzed using ImageJ software.

Varalda M., Antona A., Bettio V., Roy K., Vachamaram A., Yellenki V., Massarotti A., Baldanzi G, & Capello D. (2020). Psychotropic Drugs Show Anticancer Activity by Disrupting Mitochondrial and Lysosomal Function. Frontiers in Oncology, 10, 562196.

+ Open protocol

+ Expand

Western Blot Analysis of Liver Protein Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Liver homogenates and cells were lysed in ice-cold RIPA buffer (50 mM Tris HCl (pH 7.4), 250 mM NaCl, 1% Nonidet P-40, and protease inhibitor cocktail). The protein samples (50 µg) were separated by 4%–12% sodium dodecyl sulfate polyacrylamide gel electrophoresis and then transferred onto PVDF membranes. After blocking with 5% BSA or 5% non-fat dry milk, the membranes were incubated overnight with the following primary antibodies: anti-p-mTOR, anti-mTOR, anti-p-p70s6kinase, anti-p70s6kinase, anti-p-4EBP-1, anti-4EBP-1, anti-CHOP, anti-p-eIF2α, and anti-eIF2α (Cell Signaling Technology, Boston, MA, USA), anti-ubiquitin, anti-GRP78, anti-p-PERK, anti-PERK, anti-Bax, anti-lamp-1, anti-Tom20, anti-cathepsin B, and β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-p62 (SQSTM1; MBL International, Woburn, MA, USA), and anti-LC3II (Novus Biologicals, Littleton, CO, USA). Immunoreactive bands were visualized using the ECL Western blotting protocol (Bio-Rad). Finally, the membranes were exposed to imaging film (Kodak BioFlexEcono Scientific Supplies, Citrus Heights, CA, USA), after which the film was developed using a Kodak X-OMAT 1000A Processor. Densitometric analysis of the bands was conducted using the Image J software (NIH, Bethesda, MD, USA).

Lee G.H., Lee H.Y., Park S.A., Shin T.S, & Chae H.J. (2019). Eucommia ulmoides Leaf Extract Ameliorates Steatosis Induced by High-fat Diet in Rats by Increasing Lysosomal Function. Nutrients, 11(2), 426.

+ Open protocol

+ Expand

Isolation and Enrichment of Murine Lysosomes

Check if the same lab product or an alternative is used in the 5 most similar protocols

The isolation of lysosomes from murine tissue was performed according to
Fritsch et al.⁴⁷. In brief,
tissue was thawed in 500 μl of homogenization buffer (HB) (15 mM HEPES
(pH 7.4), 250 mM sucrose, 0.5 mM MgCl₂ containing complete protease
inhibitor and cyanase nuclease (SERVA Electrophoresis)) followed by three rounds
of careful sonication (10 s, amp 2.5 at 4 °C using a cooled cup resonator
(G. Heinemann) in a total volume of 1 ml) and centrifugation for 4 min at
1,500g. The resulting supernatant was loaded on a 16%
iodixanol-HB cushion and centrifuged for 1 h at 150,000g. The
resulting floating fraction was aspirated carefully from the cushion and diluted
to 750 μl with HB. Two micrograms of anti-Lamp1 (Santa Cruz
Biotechnology, cat. no. sc-8098) was added and samples were incubated for 30 min
while rolling at 4 °C. Five microliters of Protein G microbeads (Miltenyi
Biotec, cat. no. 130–071-101) were added followed by another 30-min
incubation. Samples were then loaded onto a HOKImag magnetic isolation device
(Hoock) for organelle pulldown. The eluate was finally sedimented by
centrifugation for 1 h at 20,000g and used for downstream
application.

Melum E., Jiang X., Baker K.D., Macedo M.F., Fritsch J., Dowds C.M., Wang J., Pharo A., Kaser A., Tan C., Pereira C.S., Kelly S.L., Duan J., Karlsen T.H., Exley M.A., Schütze S., Zajonc D.M., Merrill A.H., Schuchman E.H., Zeissig S, & Blumberg R.S. (2019). Control of CD1d-restricted antigen presentation and inflammation by sphingomyelin. Nature immunology, 20(12), 1644-1655.

+ Open protocol

+ Expand

Visualizing Rab11, EEA-1, and Lamp-1 Localization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hela cells, which expressed EGFP-SH3BP5 or EGFP-SH3BP5L transiently, were cultured on coverslips and then washed with PBS twice. The cells were fixed with 4% paraformaldehyde in PBS for 15 min and washed three times. Then, we permeabilized cells with 0.05% saponin and incubated them with PBS containing 5% FBS for 30 min for blocking and treated with the following antibodies. We used anti-Rab11 (1:250 dilution; #71-5300; Invitrogen), anti-EEA-1 (1:250 dilution; #610456; BD Biosciences), and anti-Lamp-1 (1:250 dilution; #sc-20011; Santa Cruz Biotechnology) antibodies as the primary antibodies and goat anti-rabbit or anti-mouse Alexa Fluor 594 (1:1,000; Life Technologies Inc.). We acquired images using an FV1200 confocal microscope (Olympus) with a 100× PlanApo oil immersion lens (1.40 numerical aperture; Olympus). For mitochondria staining, we incubated Hela cells, which expressed EGFP-SH3BP5 or EGFP-SH3BP5L transiently in glass bottom dishes, with a MitoTracker Red CMXRos (0.1 nM; M7512; Invitrogen) for 15 min and washed them with PBS twice for observation.

Goto-Ito S., Morooka N., Yamagata A., Sato Y., Sato K, & Fukai S. (2019). Structural basis of guanine nucleotide exchange for Rab11 by SH3BP5. Life Science Alliance, 2(2), e201900297.

+ Open protocol

+ Expand

Immunofluorescence Assay Protocol for Cellular Localization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence assays were performed as reported [50 (link)]. Specifically, primary anti-gliadin (Abcam, Cambridge, UK) and anti-Lamp1 (Santa Cruz Biotechnology, Dallas, TX, USA) antibodies were diluted 1:30 in T-PBS 5% (v/v) and incubated for one hour at room temperature. Species-specific Alexa Fluor 488 and Alexa Fluor 633 (Molecular Probes, Eugene, OR, USA) secondary conjugated antibodies (1:100 in T-PBS 5%) were then incubated for an additional one hour at room temperature. Cells grown on coverslips were washed three times with PBS and fixed with 4% (v/v) paraformaldehyde-PBS, pH 7.4, for 15 min at room temperature. Coverlips were then washed three times with PBS and were treated with ProLong Gold antifade reagent with DAPI (Invitrogen), according to the manufacturer’s instructions and finally mounted onto microscope glass slides. Fluorescence signals were detected using a fluorescence light inverted microscope (Nikon Eclipse TS100, Tokio, Japan) with a Plan Fluor 100× oil immersion objective.

Manai F., Azzalin A., Gabriele F., Martinelli C., Morandi M., Biggiogera M., Bozzola M, & Comincini S. (2018). The In Vitro Effects of Enzymatic Digested Gliadin on the Functionality of the Autophagy Process. International Journal of Molecular Sciences, 19(2), 635.

+ Open protocol

+ Expand

Immunofluorescence Staining of Cell Organelles

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were grown on cover glasses for 1 day, rinsed with TBS at room temperature and fixed for 10 min with 4% paraformaldehyde. After rinsing with TBS, in the case of permeabilized conditions, cells were incubated with 0.1% Triton X-100 in TBS for 30 min followed by 30 min in blocking buffer (BSA 2% in TBS-T 0.1% Triton X-100) at room temperature. For non-permeabilized conditions, cells were directly incubated in blocking buffer (BSA 2% in TBS) for 1 h at room temperature. Next, cells were incubated in the suitable primary antibodies for 1 h at room temperature and rinsed repeatedly with TBS before incubating in the appropriate fluorescein-labelled secondary antibody for 1 h at room temperature. Cells were then washed extensively with TBS and mounted on a glass slide in Prolong Diamond Antifade mountant with DAPI (Pierce, Thermo Fisher Scientific). The following primary antibodies were used: anti-EEA1 (1:1000) (Cell Signalling), anti-LAMP-1 (1:1000) (Santa Cruz), anti-avb3 (1:50) clone LM609 (Merck), and anti-tubulin (1:1000) (DM1a-FTIC, Sigma). The following secondary antibodies were used: goat anti-mouse IgG (H + L) Alexa Fluor Plus 488 and goat anti-rabbit IgG (H + L) Alexa Fluor 633 (Thermo Fisher Scientific) (1:1000).

Fuentes P., Sesé M., Guijarro P.J., Emperador M., Sánchez-Redondo S., Peinado H., Hümmer S, & Ramón y. Cajal S. (2020). ITGB3-mediated uptake of small extracellular vesicles facilitates intercellular communication in breast cancer cells. Nature Communications, 11, 4261.

+ Open protocol

+ Expand

Platelet Morphology Assessment via Microscopy

Check if the same lab product or an alternative is used in the 5 most similar protocols

Platelet morphology in the Spanish index was assessed by light and immunofluorescence microscopy on blood smears as reported [46 (link),47 (link)]. Particularly, the expression of three α-granule markers (i.e., thrombospondin, von Willebrand factor (vWF), and P-selectin), three dense granule markers (i.e., markers LAMP-1, LAMP-2, and CD63), and the cytoskeletal protein ß-tubulin was determined. The following primary antibodies were used: anti-thrombospondin (ab85762, Abcam, Cambridge, UK); anti-P-selectin (555522, BD Biosciences, San Jose, CA, USA); anti-vWF (A0082, Dako, Waldbronn, Germany); anti-LAMP-1 (sc18821, Santa Cruz Biotechnology, Heidelberg, Germany); anti-LAMP-2 (sc18822, Santa Cruz, CA, USA); anti-CD63 (558019, BD Biosciences); anti-β1-tubulin (T4026, Merck Life Science, Darmstadt, Germany). As secondary antibodies, Alexa Fluor 568 (A11011, Invitrogen, Thermo Fisher Scientific) and Alexa Fluor 488 (A11001, Invitrogen, Thermo Fisher Scientific, Dreieich, Germany) were used. Each marker was eventually assessed by standard immunofluorescence microscopy using the Olympus BX40 microscope system (Olympus, Hamburg, Germany). Blood smears from healthy controls were stained and analyzed in parallel.

Bastida J.M., Malvestiti S., Boeckelmann D., Palma-Barqueros V., Wolter M., Lozano M.L., Glonnegger H., Benito R., Zaninetti C., Sobotta F., Schilling F.H., Morgan N.V., Freson K., Rivera J, & Zieger B. (2022). A Novel GATA1 Variant in the C-Terminal Zinc Finger Compared with the Platelet Phenotype of Patients with A Likely Pathogenic Variant in the N-Terminal Zinc Finger. Cells, 11(20), 3223.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!