HEK293T cells were transiently cotransfected with pCAGGS-E2-Flag (1 μg) and pMyc-MEK2 (1 μg). PK-15 cells were mock infected or infected with CSFV at an MOI of 0.1. At 36 hpt or 48 hpi, the plasmid-transfected or CSFV-infected cells were fixed with 4% paraformaldehyde and permeabilized with 0.15% Triton X-100. The cells were further incubated with a mouse anti-Flag MAb (catalog no. F1804; Sigma-Aldrich) or anti-E2 MAb HQ06 (31 (link)) for 1.5 h, followed by incubation with a rabbit anti-Myc PAb (catalog no. C3956; Sigma-Aldrich) or a rabbit anti-MEK2 PAb (catalog no. sc-525; Santa Cruz). Subsequently, the cells were incubated with fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG antibody (catalog no. F2012; Sigma-Aldrich) and tetramethyl rhodamine isocyanate-conjugated goat anti-rabbit IgG antibody (whole molecule) (catalog no. T6778; Sigma-Aldrich). After incubation with 4,6-diamidino-2-phenylindole (DAPI), the cells were observed using a Leica SP2 confocal system (Leica Microsystems; Germany).
Rabbit anti myc pab
Manufactured by Merck Group
Sourced in China
Rabbit anti-Myc PAb is a primary antibody produced in rabbits that specifically recognizes the Myc protein. It is designed for use in various immunological techniques to detect and study the Myc protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti flag mab, by Merck Group (3 mentions)
Fluorescein isothiocyanate fitc conjugated goat anti mouse igg antibody, by Merck Group (1 mentions)
Rabbit anti flag pab, by Merck Group (1 mentions)
Alexa fluor 488 conjugated goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions)
Mouse anti myc mab, by Merck Group (1 mentions)
Humidified incubator, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Mouse anti β actin monoclonal antibody mab, by Merck Group (1 mentions)
Mouse anti ha mab, by Merck Group (1 mentions)
4 protocols using rabbit anti myc pab
1
Immunofluorescence Assay for E2 and MEK2 Proteins
2
Antibody Production and Characterization Protocol
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Chicken antisera against A/chicken/Shanghai/S1053/2013(H7N9) and a mouse anti-NP MAb were prepared in our laboratory by using conventional methods. The following primary antibodies were purchased from commercial sources: rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) polyclonal antibody (PAb) (product no. 10494-1-AP from Proteintech, Wuhan, China), mouse anti-Flag MAb (product no. F3165), mouse anti-Myc MAb (product no. M4439), rabbit anti-Flag PAb (product no. F7425), and rabbit anti-Myc PAb (product no. C3965) from Sigma-Aldrich. The secondary antibody used for Western blotting was DyLight 800-conjugated goat anti-rabbit IgG (H+L) (product no. 072-07-15-06), purchased from KPL (Gaithersburg, MD); the secondary antibody used for confocal microscopy was Alexa Fluor 488-conjugated goat anti-mouse IgG (H+L) (product no. A11029), obtained from Thermo Fisher Scientific (Waltham, MA).
3
PRRSV Infection in Cell Lines
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MARC-145 and 293FT cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco). Porcine pulmonary alveolar macrophages (PAMs) were prepared as previously described (38 (link)), and cultured in RPMI 1640 medium containing 10% FBS. All cells were cultured at 37°C with 5% CO2 in a humidified incubator (Thermo Fisher Scientific). The Chinese highly pathogenic PRRSV (HP-PRRSV) strain JXwn06 (GenBank accession no. EF641008 ) and low pathogenic PRRSV (LP-PRRSV) strain HB-1/3.9 (EU360130) were used in this study (39 (link)). Rabbit anti-p21 and anti-p27 polyclonal antibodies (pAb) were purchased from Proteintech Group (Chicago, IL, USA). Mouse anti-β-actin monoclonal antibody (mAb), mouse anti-HA mAb, rabbit anti-Myc pAb, mouse anti-Flag mAb were all purchased from Sigma-Aldrich. The specific mAbs raised against the N and nsp11 proteins of PRRSV were prepared in our laboratory. The plasmid pcDNA3.1-HA-Ubiquitin was purchased from Bioworld Technology, Inc. (Bloomington, MN, USA).
4
Exploring GBP1 Interaction with FMDV NS5A
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BHK-21 cells grown to 60% confluence were cotransfected with pMyc-NS5A and either pFlag-GBP1 or pFlag-GBP1(K51A) (2 μg each), and PK-15 cells treated with 100 ng of IFN-β were infected with Shimen at an MOI of 0.1 for 48 h. The transfected or infected cells were fixed with 4% paraformaldehyde, blocked with 5% skim milk for 2 h, and then the transfected cells were incubated with a mouse anti-Flag MAb (1:100) or a rabbit anti-Myc PAb (1:100) (catalog no. C3956; Sigma-Aldrich), and the infected cells were incubated with a mouse anti-NS5A PAb (1:100) or a rabbit anti-GBP1 PAb (1:200) (catalog no. ab121039; Abcam) for 1 h. Following 1 h of incubation with an Alexa Fluor 647-conjugated donkey anti-mouse IgG(H+L) antibody (catalog no. 1692912; Life Technologies) and an Alexa Fluor 488-labeled donkey anti-rabbit IgG(H+L) antibody (catalog no. 1674651; Life Technologies), the cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) for 10 min and examined using a Leica SP2 confocal system (Leica Microsystems). The colocalization coefficients were calculated by professional quantitative colocalization analysis software (CoLocalizer Pro, version 2.7.1).
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