Immunohistochemical staining was performed as previously described (Miao et al. 2020 (link)). Briefly, coronal brain sections were blocked with 5% goat serum in phosphate-buffered saline with 0.1% Triton-X 100 for 1 h, followed by primary antibody incubations for 1 h at room temperature and overnight incubation at 4 °C. The following primary antibodies were used: Rat anti MAP2 (1:400; Millipore), Rabbit anti Ly6G (1:300; Abcam), Rabbit anti F4/80 (1:300; Bio Legend), Alexa Flour 488-conjugated (1:300; Invitrogen), Rat anti CD31 (1:300; BD Biosciences). Rabbit anti-MMP-2 (1:200, ab97779, Abcam), goat anti-CD206 (1:250, AF2535, R&D), rat anti-CD16/32 (1:250, ab25235, Abcam), and rabbit anti-Iba1 (1:1000, ab5076, Abcam).
Alexa flour 488 conjugated
Manufactured by Thermo Fisher Scientific
Alexa Fluor 488-conjugated is a fluorescent dye manufactured by Thermo Fisher Scientific. It is designed for use in various fluorescence-based applications, such as cellular imaging, flow cytometry, and immunoassays. The dye is characterized by its bright green fluorescence and can be excited by a 488 nm light source.
Automatically generated - may contain errors
Lab products found in correlation
Human tumor necrosis factor α tnf α 8902, by Cell Signaling Technology (2 mentions)
Ab97779, by Abcam (1 mentions)
Ab25235, by Abcam (1 mentions)
Ab5076, by Abcam (1 mentions)
Rat anti cd31, by BD (1 mentions)
Rabbit anti ly6g, by Abcam (1 mentions)
Af2535, by R&D Systems (1 mentions)
Peroxidase conjugated affinipure goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Anti flag mab f3165, by Merck Group (1 mentions)
Anti flag pab f7425, by Merck Group (1 mentions)
Anti gfp pab, by Thermo Fisher Scientific (1 mentions)
Peroxidase conjugated affinipure goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Alexa flour 568 conjugated, by Thermo Fisher Scientific (1 mentions)
Polyinosinic polycytidylic poly 1 c, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
Minimum essential medium (mem), by Corning (1 mentions)
4 protocols using alexa flour 488 conjugated
1
Immunohistochemical Staining of Brain Sections
2
Anti-PRRSV nsp1β Rabbit Antibody Generation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An anti-PRRSV-nsp1β rabbit pAb specific for NA-PRRSV nsp1β was generated at the Immunological Research Center, University of Illinois at Urbana-Champaign (Urbana, IL). The anti-LV-nsp2/3 antibody was a generous gift from E. Snijder, (Leiden University Medical Center, Leiden, Netherlands). Anti-N protein mAbs (SDOW17) was obtained from E. Nelson (South Dakota State University, Brookings, SD). polyinosinic: polycytidylic [poly (I:C)], DAPI (4′,6-diamidino-2-phenylindol), anti-Flag mAb (F3165), and anti-FLAG pAb (F7425) were purchased from Sigma (St. Louis, MO). Anti-β-actin mAb (sc-47778), anti- HSP90 mAb (sc-69703), and anti-PARP pAb (sc-7150) were purchased from Santa Cruz Biotechnologies Inc. (Santa Cruz, CA). Anti-ISG15 pAb and anti-GFP pAb were purchased from Thermo Scientific (Rockford, IL). Anti-DEPTOR pAb was purchased from Invitrogen (Carlsbad, CA). The peroxidase-conjugated Affinipure goat anti-mouse IgG and peroxidase-conjugated Affinipure goat anti-rabbit IgG were purchased from Jackson Immuno Research (West Grove, PA). Alexa-Flour 488-conjugated and Alexa-Flour 568-conjugated secondary antibodies were purchased from Invitrogen.
3
PRRSV Nsp1β Antibody Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies and chemicals used in the present study are listed as follows. Anti (α)-PRRSV-nsp1β PAb (rabbit polyclonal antibody) specific for PRRSV-2 nsp1β was generated at the Immunological Research Center, University of Illinois at Urbana-Champaign (Urbana, IL). α-N protein MAb (MR40; mouse monoclonal antibody) was obtained from E. Nelson (South Dakota State University, Brookings, SD). α-p65 Mab (mouse) (F-6, sc-8008) was purchased from Santa Cruz Biotechnologies Inc. (Santa Cruz, CA). Alexa-Flour 488-conjugated and Alexa-Flour 568-conjugated secondary antibodies were obtained from ThermoFisher (Rockford, IL). Human tumor necrosis factor-α (TNF-α) (8902) was purchased from Cell Signaling (Danvers, MA). DAPI (4′, 6′-diamidino-2-phenylindol) and Polyinosinic:polycytidylic [poly (I:C)] were obtained from Sigma (St. Louis, MO). Human and porcine recombinant IFN-β were purchased from Calbiochem (San Diego, CA), and for stimulation, 1000 unit/ml was added to cells for 6 h.
4
Immunofluorescence Assay for NF-κB Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa (NIH AIDS Research and Reference Reagent Program, Germantown, MD) cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Corning Inc., Corning, NY), supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco, Grand Island, NY), in a humidified incubator with 5% CO2 at 37 °C. A549 cells and 16HBE14o cells (Kindly provided by Gee Lau, University of Illinois) were maintained in Minimum Essential Medium (MEM; Corning Inc., Corning, NY), supplemented with 10% heat-inactivated FBS, in a humidified incubator with 5% CO2 at 37 °C. Anti (α)-FLAG PAb (rabbit) was purchased from Rockland Inc (Gilbertsville, PA). α-p65 Mab (mouse) (F-6, sc-8008) was purchased from Santa Cruz Biotechnologies Inc. (Santa Cruz, CA). Alexa-Flour 488-conjugated, and Alexa-Flour 568-conjugated secondary antibodies were obtained from ThermoFisher (Rockford, IL). Human tumor necrosis factor-α (TNF-α) (8902) was purchased from Cell Signaling (Danvers, MA). DAPI (4′, 6′-diamidino-2-phenylindol) was obtained from Sigma (St. Louis, MO).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!