Anti clathrin

Manufactured by Abcam

Sourced in United Kingdom, Switzerland

Anti-clathrin is a laboratory reagent used to detect and study the protein clathrin, which is involved in the formation of clathrin-coated vesicles during cellular processes such as endocytosis. It functions as a specific binding agent for clathrin molecules.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) General chemicals, by Merck Group (1 mentions) Phospho erk1 2, by Cell Signaling Technology (1 mentions) Phospho akt ser473, by Cell Signaling Technology (1 mentions) Alexa fluor 647 phalloidin, by Thermo Fisher Scientific (1 mentions) Neutravidin protein, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Anti dynamin 2, by BD (1 mentions) Horseradish peroxidase hrp conjugated anti rabbit igg, by Cell Signaling Technology (1 mentions) Hrp conjugated anti mouse igg, by Cell Signaling Technology (1 mentions) Las4000, by GE Healthcare (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Rabbit igg blocking reagent, by Thermo Fisher Scientific (1 mentions) Anti egf receptor, by Cell Signaling Technology (1 mentions) Rabbit anti homer, by Synaptic Systems (1 mentions) Rabbitanti rab5 antibody, by Cell Signaling Technology (1 mentions) Kt5720, by Merck Group (1 mentions) Pkc inhibitor go 6976, by Merck Group (1 mentions) Alexa flour conjugated phalloidin, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Anti-clathrin heavy chain Rabbit anti-clathrin Rabbit anti-clathrin heavy chain Clathrin

5 protocols using anti clathrin

Fullerene and SWCNT Characterization

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General chemicals were from VWR (West Chester, PA) and Sigma (St. Louis, MO). Phorbol 12,13-myristate acetate (PMA) and ionomycin were from Calbiochem (Gibbstown, NJ). Antibodies were sourced as follows: phospho-p72syk, phospho-ERK1/2, phospho-AKT (Ser 473) were from Cell Signaling Technologies (Danvers, MA). Anti-phospho-phospholipase C gamma 1 was from BD Biosciences (San Jose, CA). Anti-phospho-NFATC1 (Ser54) and anti-clathrin were from Abcam (Cambridge, MA). Anti-Grb2 was from Upstate Biotechnologies (Billerica, MA). Alexa-conjugated wheat germ agglutinin (WGA), Alexa- and HRP conjugated secondary antibodies were from Invitrogen (Temecula, CA) and Amersham (Piscataway, NJ).
Water-soluble hydroxylated fullerene derivatives (C₆₀(OH)_n, polyhydroxy C-60 modified with 18-22 hydroxyls) were obtained from BuckyUSA (Houston, TX). SWCNT were produced by CVD at >95% purity and <5% Fe contamination (Nano-Labs, Detroit, MI). Note that DMEM contains ∼0.0001g/L FeNO₃.9H₂O. TEM shows a SWCNT width range of 10-21 nm and a mixture of agglomerated and single SWCNT with lengths ranging from <2 to >10 microns. Measured aggregate sizes interacting with cells ranged from ∼100nm to >1 micron diameter. Doses of SWCNT from 10-100 μg/ml were calculated to equate to 2.8-28 micrograms per cm² in microscopy applications.

Umemoto E.Y., Speck M., Shimoda L.M., Kahue K., Sung C., Stokes A.J, & Turner H. (2014). Single-walled carbon nanotube exposure induces membrane rearrangement and suppression of receptor-mediated signalling pathways in model mast cells. Toxicology letters, 229(1), 198-209.

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Antibody Labeling and Cell Culture Protocols

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HeLa cells were cultured in Dulbecco’s Modified Eagle Medium (Life Technologies # 10313–021) supplemented with 10% fetal bovine serum (HyClone), penicillin-streptomycin, and 2 mM L-glutamine, and cells were maintained at sub-confluence.
Anti-EGFR-AF647 antibody (anti-EGFR, R-1, sc-101) was purchased from Santa Cruz Biotechnology. Anti-alpha-tubulin (T6074) and anti-beta-tubulin (T8328) antibodies were purchased from Sigma. Anti-clathrin was purchased from Abcam (Clathrin heavy chain, ab21679). All primary antibodies were either purchased pre-conjugated to AF647 or were conjugated directly (see below) using AlexaFluor647 carboxylic acid, succinimidyl ester (A-20006), purchased from Life Technologies, with the exception of Anti-clathrin, which was directly conjugated using the APEX AlexaFluor-647 antibody labeling kit (Life Technologies, A10475) due to the presence of carrier proteins. AlexaFluor-647 phalloidin was purchased from Life Technologies (A22287). Sodium borohydride was purchased from EMD (SX0380-3). Photocleavable biotin NHS (PC-biotin-NHS) was purchased from Ambergen, and purified NeutrAvidin protein was purchased from ThermoScientific (31000).

Valley C.C., Liu S., Lidke D.S, & Lidke K.A. (2015). Sequential Superresolution Imaging of Multiple Targets Using a Single Fluorophore. PLoS ONE, 10(4), e0123941.

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Protein Expression Analysis of Cells Treated with SBHA and MS-275

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Cells were treated with SBHA (20 μM) or MS-275 (2 μM) for 24 h. Cells were lysed in NP-40 (Elpisbio, Daejeon, Korea) in the presence of a protease inhibitor cocktail (Sigma-Aldrich). Protein concentration was determined using the bicinchoninic acid protein assay (Pierce Rockford, IL Waltham, MA, USA). Each sample was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto a polyvinyl difluoride (PVDF) membrane. Membranes were incubated with anti-dynamin-2 (BD Bioscience), anti-clathrin (Abcam, Cambridge, UK), and anti-β-actin (Santa Cruz) Abs then incubated with either horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (Cell signaling technology, Beverly, MA, USA) or HRP-conjugated anti-mouse IgG (Cell signaling technology) secondary Abs. Blots were visualized using a super-signal chemiluminescence kit (Pierce, Rockford, IL, USA) and developed using image quant LAS 4000 (GE Lifesciences, Pittsburgh, PA, USA).

Chang H.G., Choi Y.H., Hong J., Choi J.W., Yoon A.R, & Yun C.O. (2021). GM101 in Combination with Histone Deacetylase Inhibitor Enhances Anti-Tumor Effects in Desmoplastic Microenvironment. Cells, 10(11), 2811.

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Immunofluorescence Imaging of Clathrin and EGFR

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Cells were fixed 4 % paraformaldehyde in PBS for 20 min, permeabilized with 0.1 % Triton X-100 in PBS for 5 min, then washed in PBS, then in PBS supplemented with 1% BSA and 1% rabbit IgG blocking reagent (ThermoFisher) for 5 min, then in PBS. Anti-Clathrin (Abcam, ab21679) and anti-EGF Receptor (Cell Signaling, D38B1) were labelled with Alexa Fluor 488-and Alexa Fluor 647-Zenon Rabbit IgG labelling kits respectively (ThermoFisher), according to the manufacturer's instructions, and cells were then incubated with both antibodies (both diluted 1:100 in PBS-1% BSA) for 20 minutes. After washing thrice in PBS, imaging was performed in PBS instead of mounting medium to avoid squashing the cells (and potentially biasing the Clathrin or EGFR / pattern colocalization).

Watson J.L., Aich S., Oller‐Salvia B., Drabek A.A., Blacklow S.C., Chin J.W., & Derivery E. (2020). Fibrinogen anchors for micropatterning of active proteins and subcellular receptor relocalisation.

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HA-tagged DAGLα Construct Generation

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Rat or mouse anti-HA antibodies were from Roche (Roche, Switzerland); Rabbit anti-HA, anti-V5, anti-clathrin, anti-early endosome antigen 1 (EEA1) and anti-lysosomalassociated membrane protein 1 (LAMP1) antibodies were from Abcam (Abcam, UK); Rabbit anti-Homer was from Synaptic Systems (Synaptic Systems, Goettingen, Germany); Rabbit anti-Rab5 antibody was from Cell Signaling (Massachusetts, USA).
Alexa Flour conjugated phalloidin, transferrin and secondary antibodies were from Life Technologies. PKC inhibitor Go 6976, PKA inhibitors H89 and KT 5720 were from Sigma (Sigma-Aldrich, UK). For HA-DAGLα construct, a HA peptide sequence (YPYDVPDYA) flanked by a glycine-serine-glycine-serine linker on either side was inserted into the first extracellular loop of Human DAGLα sequence in pcDNA™6.2-DEST/V5 construct (performed by GenScript, Piscataway, USA).

Zhou Y., Howell F.V., Glebov O.O., Albrecht D., Williams G, & Doherty P. (2016). Regulated endosomal trafficking of Diacylglycerol lipase alpha (DAGLα) generates distinct cellular pools; implications for endocannabinoid signaling. Molecular and cellular neurosciences, 76.

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