Calreticulin

Manufactured by Merck Group

Sourced in Spain

Calreticulin is a protein that functions as a calcium-binding chaperone in the endoplasmic reticulum of eukaryotic cells. It plays a role in the folding and assembly of newly synthesized glycoproteins.

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Lab products found in correlation

Gm130, by BD (2 mentions) γ tubulin, by Abcam (1 mentions) α tubulin, by Merck Group (1 mentions) Clone 20h5, by Merck Group (1 mentions) G7295, by Merck Group (1 mentions) Prolong gold, by Thermo Fisher Scientific (1 mentions) Mounting medium, by Ibidi (1 mentions) Cdc42, by BD (1 mentions) Fibronectin, by Roche (1 mentions) Lamp1, by Merck Group (1 mentions) Goat anti mouse alexa 488, by Thermo Fisher Scientific (1 mentions) Fv1000 confocal laser scanning microscope, by Olympus (1 mentions)

Close spelling variants of this product

Anti-calreticulin (4 citations)

3 protocols using calreticulin

Immunofluorescence Staining of Cell Markers

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Primary antibodies to the following proteins were used in this study: calreticulin (C4606; Sigma-Aldrich), Tuba (a gift from Frank Gertler, MIT; and B01P, Abnova), centrin2 (a gift from Jeffery L. Salisbury, Mayo Clinic; and Clone 20H5, Millipore), FGD1 (HPA 000911; Sigma-Aldrich), myc (Clone 9E10; Cal Biochem), GM130 (Clone 35; BD Biosciences; and G7295, Sigma-Aldrich), giantin (a gift from Vivek Malhotra, Center for Genomic Regulation, Barcelona, Spain), kendrin (a gift from Mikiko Takahashi, Teikyo Heisei University), α-tubulin (T5168; Sigma-Aldrich), γ-tubulin (ab11310; Abcam), Cdc42 (Clone 44; BD Biosciences), and vesicular stomatitis virus G-protein (VSVG; BWG85; a gift from Victor Hsu, Harvard Medical School). Secondary antibodies for immunofluorescence were from Theromo Fisher or Biotium, and near-infrared antibodies for Western blots were from LI-COR (Lincoln, NE). For immunofluorescence, cells were fixed in either 100% ice-cold methanol (JT Baker) or 4% formaldehyde (Ted Pella), blocked and permeabilized with 2% blocking buffer (2% fetal bovine serum [FBS], 0.01% Triton X-100, and 1× phosphate-buffered saline), stained with primary antibodies for 1 h at room temperature, and stained with secondary antibodies for 1 h. Coverslips were mounted with ProLong Gold (Thermo Fisher), and imaging dishes were filled with Ibidi Mounting Medium (Ibidi).

Herrington K.A., Trinh A.L., Dang C., O’Shaughnessy E., Hahn K.M., Gratton E., Digman M.A, & Sütterlin C. (2017). Spatial analysis of Cdc42 activity reveals a role for plasma membrane–associated Cdc42 in centrosome regulation. Molecular Biology of the Cell, 28(15), 2135-2145.

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Imaging Monocyte Subcellular Localization

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Fibronectin-coated coverslips were made by incubation of 20 μg/mL Fibronectin (Roche) in PBS for 1 h at 37 °C. Monocytes were adhered on Fibronectin-coated coverslips for 2 h and subsequently fixed with 2 % paraformaldehyde (PFA) and blocked with 3 % bovine serum albumin (BSA), 1 % HS and 10 mM glycine in PBS for 30 min at room temperature (RT). Cells were permeabilized and stained with antibodies against CD53 (mem53, Serotec), CD37 (WR17, home-made), calreticulin (ER marker, Sigma), syntaxin 12/13 [endosome marker, Synaptic Systems (cat. no. 110132)] and Lamp1 (lysosome marker, Sigma-Aldrich) in 0.5 % saponin, 1 % BSA, 10 mM glycine, 1 % HS in PBS, followed by goat-anti-mouse Alexa488 and goat-anti-rabbit Alexa647 (Molecular Probes). Samples were imaged with an Olympus FV1000 confocal laser scanning microscope. Images were analyzed using Fiji software (Schindelin et al. 2012 (link)).

de Winde C.M., Zuidscherwoude M., Vasaturo A., van der Schaaf A., Figdor C.G, & van Spriel A.B. (2015). Multispectral imaging reveals the tissue distribution of tetraspanins in human lymphoid organs. Histochemistry and Cell Biology, 144(2), 133-146.

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Protein Localization in HeLa Cells

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A T-75 flask of near-confluent HeLa cells was transfected with 12 μg of pEGFP, pGFP-Ank9, pGFP-Ank9ΔF-box, or pGFP-Ank9_47–422. A confluent flask of untransfected HeLa cells was used as a control. At 4 h or 16 h post transfection, the cells were washed twice with cold PBS prior to processing for density gradient centrifugation and trichloroacetic acid precipitation as previously described (Truchan et al., 2016 (link)). Equal volumes of fractions 1 to 9 were immunoblotted using GM130 (1:500, BD Biosciences), calreticulin (1:4000, Sigma Aldrich), and GFP (1:1000, Thermo Fisher) antibodies.

Beyer A.R., Rodino K.G., VieBrock L., Green R.S., Tegels B.K., Oliver LD J.r., Marconi R.T, & Carlyon J.A. (2017). Orientia tsutsugamushi Ank9 is a multifunctional effector that utilizes a novel GRIP-like Golgi localization domain for Golgi-to-endoplasmic reticulum trafficking and interacts with host COPB2. Cellular microbiology, 19(7), 10.1111/cmi.12727.

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