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Goat anti rabbit igg alexa fluor 555 a21428

Manufactured by Thermo Fisher Scientific

Goat anti-rabbit IgG Alexa Fluor 555 (A21428) is a secondary antibody conjugated with the Alexa Fluor 555 fluorescent dye. It is designed to detect and visualize rabbit primary antibodies in various immunoassay applications.

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2 protocols using goat anti rabbit igg alexa fluor 555 a21428

1

SARS-CoV-2 Infection Assay in Calu-3 and Vero E6 Cells

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Calu-3 cells52 (link) (ATCC HTB-55) were cultured according to ATCC recommendations. All experiments were performed in these cells below passage 15. Vero E6 cells (ATCC CRL-1586; used for SARS-CoV-2 plaque assay) were grown in minimum essential medium (MEM) supplemented with 10% FBS, 1 mM sodium pyruvate and 0.1 nM non-essential amino acids, and used at passage <40. All cells were expanded in a T175 flask with 5% carbon dioxide at 37 °C. Cell density was kept between 0.25 and 2 million cells per ml. Vero E6 cells (used for TMPRSS2 proteolytic activity screening and IC50 determination) were maintained in Eagle’s minimum essential medium (EMEM) containing 10% foetal bovine serum, 2 mmol l−1l-glutamine, 100 IU ml−1 penicillin and 100 μg ml−1 streptomycin. Cm was obtained from MilliporeSigma. The SARS-CoV-2 nucleocapsid antibody (HL344) (GTX635679) was provided by Genetex; mouse anti-dsRNA antibody (J2-1904) was purchased from Scions English and Scientific Consulting33 (link); Hoechst 33342 and secondary antibodies goat anti-mouse IgG Alexa Fluor 488 (A11001) and goat anti-rabbit IgG Alexa Fluor 555 (A21428) were obtained from Invitrogen. Cell lines were screened for mycoplasma contamination using MycoAlert Mycoplasma Detection Kit (Lonza).
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2

SARS-CoV-2 Cell Culture and Antibody Evaluation

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Human Calu-3 cells (ATCC® HTB-55) and human Caco-2 cells (ATCC® HTB-37) were cultivated according to ATCC recommendations. All experiments were performed in these cells below passage 15. Vero E6 cells (ATCC® CRL-I1586™; used for generating SARS-CoV-2 stocks) were cultivated in MEM that was supplemented with 10% FBS, 1 mM sodium pyruvate, and 0.1 nM non-essential amino acids and used at passage <40. VeroE6/TMPRSS2 cells (JCRB 1819) were maintained in MEM supplemented with 5% FBS and penicillin and streptomycin (100 IU and 100 µg/ml, respectively). Normal human bronchial epithelial cells (NHBE), isolated from the epithelial lining of airways above the bifurcation of the lungs (NHBE-Bronchial Epi Cells; Lonza CC-2540S) were cultivated according to Lonza recommendations. The SARS-CoV-2 nucleocapsid antibody [HL344] (GTX635679) was kindly provided by GeneTex; mouse anti-dsRNA antibody (J2-1904) was purchased from Scicons English and Scientific Consulting (Long 2020); Hoechst 33,342 and secondary antibodies goat anti-mouse IgG Alexa Fluor 488 (A11001) and goat anti-rabbit IgG Alexa Fluor 555 (A21428) were obtained from Invitrogen.
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