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C4606

Manufactured by Merck Group
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Sourced in Hungary, United Kingdom

C4606 is a laboratory equipment product offered by Merck Group. It serves as a general-purpose device for various laboratory applications. The core function of C4606 is to perform tasks related to sample handling, preparation, or analysis, though the specific intended use is not elaborated on in this factual description.

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Lab products found in correlation

Ab22595, by Abcam (2 mentions) A5420, by Merck Group (2 mentions) Ab3434 100, by Abcam (2 mentions) Bd611562, by BD (2 mentions) Na934 1ml, by Merck Group (2 mentions) Ab9485, by Abcam (2 mentions) Triton x 100, by Merck Group (1 mentions) Ds qi2 camera, by Nikon (1 mentions) Donkey serum, by Merck Group (1 mentions) Paraformaldehyde solution, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Euroclone (1 mentions) Eclipse ni u microscope, by Nikon (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) A2228, by Merck Group (1 mentions) Sc 188, by Santa Cruz Biotechnology (1 mentions) Af1857, by R&D Systems (1 mentions) Sc 32233, by Santa Cruz Biotechnology (1 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Adi spa 827, by Enzo Life Sciences (1 mentions)

5 protocols using c4606

1

Immunofluorescence Imaging of Flavivirus

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Cells grown on the cover-slip were fixed for 20 min in 4% paraformaldehyde solution (Sigma) in phosphate-buffered saline (PBS, Euroclone). Cells were permeabilized for 30 min in blocking solution, containing 0.2% Triton X-100 (Sigma) and 10% donkey serum (Sigma) and incubated overnight at 4 °C with the primary mAb in blocking solution. The following mAbs specific for Flavivirus E protein (1:200, Millipore, MAB10216), double-stranded RNA (1:300, English and Scientific Consulting Kft, Hungary), vimentin (1:300, Bioss, BS-0756R) and calreticulin (1:300, Sigma, C4606) were used. Cells were then washed with PBS and incubated for 1 h with Hoechst and either anti-mouse Alexa Fluor-488 or anti-rabbit Alexa Fluor-594 secondary Abs (1:1,000 in blocking solution, ThermoFisher Scientific). High resolution wide field fluorescence images were acquired on a Nikon Eclipse Ni-U microscope equipped with a Nikon 60x plan apo 1.40 oil or a 20x plan apo 0.75 objective and a Nikon DS-Qi2 camera.
Pagani I., Ghezzi S., Ulisse A., Rubio A., Turrini F., Garavaglia E., Candiani M., Castilletti C., Ippolito G., Poli G., Broccoli V., Panina-Bordignon P, & Vicenzi E. (2017). Human Endometrial Stromal Cells Are Highly Permissive To Productive Infection by Zika Virus. Scientific Reports, 7, 44286.
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2

Comprehensive Western Blotting Antibody Assay

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Western blotting experiments were performed using the following primary antibodies: vinculin (V9131, clone hVIN-1 Sigma-Aldrich, 1:10,000), GAPDH (ab9485, Abcam, 1:1,000 or AM4300, clone 6C5, Ambion, 1:20,000), β-actin (sc-69879, clone AC-15, Santa Cruz Biotechnology, 1:2,000), p32 (A302-863A, Bethyl Laboratories, 1:5,000 or sc-10258, clone D-19, Santa Cruz Biotechnology, 1:1,000), XRN2 (A301-103A, Bethyl Laboratories, 1:2,000), CARF (A303-861A, Bethyl Laboratories, 1:2,000), NDUFS3 (Abcam 14711, 1:1,000), VDAC1 (ab3434-100, Abcam, 1:2,000), Hsp60 (BD611562, BD Biosciences, 1:5,000), Tom20 (FL-145, sc-11415, Santa Cruz Biotechnology, 1:1,000), calreticulin (C4606, Sigma-Aldrich, 1:10,000), calnexin (ab22595, Abcam, 1:5,000), SRSF1 (32-4600, Thermo Fisher Scientific, 1:250), puromycin (MABE343, clone 12D10, Merck-Millipore, 1:25,000), HA-tag (3724, clone C29F4, Cell Signaling Technologies, 1:1,000). The following HRP-linked secondary antibodies were used: anti-goat IgG (A5420, Sigma-Aldrich, 1:10,000), anti-mouse IgG (7076S, Cell Signaling Technologies, 1:5,000), anti-rabbit IgG (NA934-1ML, Sigma-Aldrich, 1:10,000).
Roberto V., Yvessa V., Hideaki I., Lucas G., Emilien N., Kritika S., Alexandros A., Riccardo D.P., Keiichi I., Pieter M., Denis L.L., Gaetano T.G., Nobuhiro T., Jean-Christophe M, & Eleonora L. (2018). SAMMSON fosters cancer cell fitness by enhancing concertedly mitochondrial and cytosolic translation. Nature structural & molecular biology, 25(11), 1035-1046.
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3

Antibody Characterization and Validation

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The following antibodies were used for immunohistochemical, immunofluorescence and Western blot analyses: sheep polyclonal antibody against uromodulin (ab9029, Abcam, Cambridge, UK) (1:200 for IF and for IHC); rat monoclonal antibody against HA (#11 867 423 001, Roche) (1:500 for IF and 1:1,000 for WB); rabbit polyclonal antibody against calreticulin (C4606, Sigma-Aldrich) (1:500 for IF); rat antibody against mouse F4/80 (MCA497GA, Serotec, BioRad)(1:50 for IF and IHC); rabbit polyclonal antibody against Atf3 (sc-188, Santa Cruz Biotechnology, Dallas, TX) (1:50 for IF and 1:500 for WB); goat polyclonal against Lipocalin 2/NGAL (AF1857, R & D Systems, Bio-Techne, Minneapolis, MN) (1:500 for WB); rabbit polyclonal against Kim1 (TIM-1) (NBP1-76701, Novus Biologicals, Bio-Techne) (1:500 for WB); mouse monoclonal antibody against β-Actin (A2228, Sigma-Aldrich) (1:20,000 for WB); mouse monoclonal antibody against Gapdh (sc-32233, Santa Cruz Biotechnology) (1:20,000 for WB).
Trudu M., Schaeffer C., Riba M., Ikehata M., Brambilla P., Messa P., Martinelli-Boneschi F., Rastaldi M.P, & Rampoldi L. (2017). Early involvement of cellular stress and inflammatory signals in the pathogenesis of tubulointerstitial kidney disease due to UMOD mutations. Scientific Reports, 7, 7383.
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4

Comprehensive Western Blotting Antibody Assay

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Western blotting experiments were performed using the following primary antibodies: vinculin (V9131, clone hVIN-1 Sigma-Aldrich, 1:10,000), GAPDH (ab9485, Abcam, 1:1,000 or AM4300, clone 6C5, Ambion, 1:20,000), β-actin (sc-69879, clone AC-15, Santa Cruz Biotechnology, 1:2,000), p32 (A302-863A, Bethyl Laboratories, 1:5,000 or sc-10258, clone D-19, Santa Cruz Biotechnology, 1:1,000), XRN2 (A301-103A, Bethyl Laboratories, 1:2,000), CARF (A303-861A, Bethyl Laboratories, 1:2,000), NDUFS3 (Abcam 14711, 1:1,000), VDAC1 (ab3434-100, Abcam, 1:2,000), Hsp60 (BD611562, BD Biosciences, 1:5,000), Tom20 (FL-145, sc-11415, Santa Cruz Biotechnology, 1:1,000), calreticulin (C4606, Sigma-Aldrich, 1:10,000), calnexin (ab22595, Abcam, 1:5,000), SRSF1 (32-4600, Thermo Fisher Scientific, 1:250), puromycin (MABE343, clone 12D10, Merck-Millipore, 1:25,000), HA-tag (3724, clone C29F4, Cell Signaling Technologies, 1:1,000). The following HRP-linked secondary antibodies were used: anti-goat IgG (A5420, Sigma-Aldrich, 1:10,000), anti-mouse IgG (7076S, Cell Signaling Technologies, 1:5,000), anti-rabbit IgG (NA934-1ML, Sigma-Aldrich, 1:10,000).
Roberto V., Yvessa V., Hideaki I., Lucas G., Emilien N., Kritika S., Alexandros A., Riccardo D.P., Keiichi I., Pieter M., Denis L.L., Gaetano T.G., Nobuhiro T., Jean-Christophe M, & Eleonora L. (2018). SAMMSON fosters cancer cell fitness by enhancing concertedly mitochondrial and cytosolic translation. Nature structural & molecular biology, 25(11), 1035-1046.
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5

Immunofluorescence Labeling of Cellular Organelles

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Cells grown on coverslip in 12-wells plate were fixed in 4% paraformaldehyde (PFA) for 20 min, permeabilised 10 min with 0.5% triton and blocked 30 min with 10% donkey serum. Cells were labelled for 1 h 30 min at room temperature with the indicated antibodies (goat polyclonal anti-uromodulin (0855140, MP Biomedicals, Santa Ana, CA, dilution 1:500); rabbit polyclonal anti-calreticulin (C4606, Sigma, dilution 1:500); mouse monoclonal anti KDEL (ADI-SPA-827, Enzo Life Sciences, Farmingdale, NY, dilution 1:500) followed by 1h incubation with the appropriate Alexa-Fluor conjugated secondary antibodies (1:500; Thermofisher). Cells were stained with 4,6-diamidino-2-phenylindole (DAPI) and mounted using FluorSave Reagent (Calbiochem, San Diego, CA). All pictures were taken with an UltraVIEW ERS spinning disk confocal microscope (UltraVIEW ERS-Imaging Suite Software, Zeiss 63X/1.4; PerkinElmer Life and Analytical Sciences Boston, MA). All images were imported in Photoshop CS (Adobe Systems, Mountain View, CA) and adjusted for brightness and contrast.
Schaeffer C., Merella S., Pasqualetto E., Lazarevic D, & Rampoldi L. (2017). Mutant uromodulin expression leads to altered homeostasis of the endoplasmic reticulum and activates the unfolded protein response. PLoS ONE, 12(4), e0175970.
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