Cryostat sections of mock and VEEV infected mice brains were incubated with a cocktail of goat anti-VEEV capsid antibody (gift from Kurt Kamrud, AlphaVax, NC) at 1:8,000 dilution with rabbit anti-Iba-1 antibody at 1:1,000 (NBP2-19019, Novus Biologicals) or rabbit anti-GFAP at 1:2,000 NB300-141, Novus Biologicals) for 2 days, after which slide sections were rinsed with PBS, and then incubated with a cocktail of Donkey anti-Rabbit IgG (H + L) Alexa Fluor 488 (Invitrogen) or Donkey anti-Goat IgG (H + L) Secondary Antibody, Alexa Fluor 594 (Invitrogen) at 1:500 overnight. Slides were rinsed with PBS counterstained with DAPI, mounted and coverslip with Fluoromount-G (SouthernBiotech, USA). Images captured with Zeiss 710 confocal microscope.
Nbp2 19019
Manufactured by Novus Biologicals
Sourced in China
NBP2-19019 is a laboratory reagent offered by Novus Biologicals. It is a product used for research purposes, but a detailed description of its core function cannot be provided in an unbiased and factual manner without potential extrapolation. Therefore, the description for this product is not available.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 donkey anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
Fluoromount g, by Southern Biotech (1 mentions)
Alexa fluor 488 antibody, by Thermo Fisher Scientific (1 mentions)
Iba1 antibody, by Novus Biologicals (1 mentions)
A1r confocal microscope, by Nikon (1 mentions)
Ab214879, by Abcam (1 mentions)
Ab254143, by Abcam (1 mentions)
Diaminobenzidine tetrahydroxyl chloride solution, by Vector Laboratories (1 mentions)
Abc kit, by Vector Laboratories (1 mentions)
Ab104224, by Abcam (1 mentions)
Ab8152, by Abcam (1 mentions)
Ab150113, by Abcam (1 mentions)
Zli 9056, by ZSGB-BIO (1 mentions)
4 protocols using nbp2 19019
1
Immunofluorescent Labeling of VEEV in Mouse Brain
2
Immunofluorescence Staining of Microglia
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Isolated microglia were seeded on poly-d -lysine–coated glass coverslips and fixed at room temperature using 4% paraformaldehyde (PFA) for 15 min. Cells were then washed twice with PBS, permeabilized, and blocked using 4% bovine serum albumin (BSA)/4% normal goat serum/0.1% Triton X-100 for 1 hour. Cells were then washed twice using PBS and incubated overnight with 1:1000 IBA1 antibody (Novus, NBP2-19019) in PBS/4% BSA at 4°C. The next day, cells were washed with PBS containing 0.1% Tween 20 and then incubated by secondary goat rabbit Alexa Fluor 488 antibody (Thermo Fisher Scientific; 1:1000 in PBS/4% BSA) at room temperature in the dark. Cells were then washed three times with PBS dissolved with 0.1% Tween 20 and mounted using ProLong Gold Antifade Mount medium with 4′,6-diamidino-2-phenylindole (DAPI; Invitrogen, P36931). Microglia were imaged on an upright Nikon A1R confocal microscope.
3
Immunohistochemical Analysis of Rat Brain
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Rat brain tissue sections were incubated in 10 mmol/L sodium citrate buffer and microwave‐treated for 20 min for antigen retrieval. After sealing with 3% H2O2 and 10% normal goat serum, the slides were incubated overnight at 4°C with mouse monoclonal antibodies against microtubule‐associated protein 2 (MAP2, 1:300, ab254143; Abcam) and IBA1 (1:200, NBP2‐19019; Novus Biologicals). The slides were then incubated with biotin‐coupled anti‐mouse secondary antibody (1:1,000, ab214879; Abcam) for 2 h at 37°C using the ABC kit (Vector Laboratories, Burlingame, CA, USA). The sections were incubated with HRP reagent, and the peroxidase activity was observed with diaminobenzidine tetrahydroxyl chloride solution (Vector Laboratories). Finally, the sections were counter‐stained with hematoxylin.
4
Immunofluorescence Staining of Brain Tissue
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Sections (20 μm) were washed in PBS and then in a microwave bath at 85–95°C for 20 min in 10 mM trisodium citrate buffer (pH 6.0). The sections were blocked in 10% normal goat serum (ZLI‐9056; ZSGB‐Bio, Beijing, China) containing 0.3% Triton X‐100 for 60 min at room temperature and incubated with antibodies against ATF3 (1:150, ab191513; Abcam), neuronal nuclear antigen (NeuN, 1:200, ab104224; Abcam), IBA1 (1:200, NBP2‐19019; Novus Biologicals), and BrdU (1:300, ab8152, Abcam) overnight at 4°C. The next day, the sections were probed with secondary antibody (1:500, ab150113; Abcam) coupled with fluorescent dye for 60 min at room temperature. Fluorescence was then detected using a fluorescence microscope (Nikon Instruments Inc., Melville, NY, USA), and the images were background subtracted using a Nikon C2 Plus (60×). NIS‐Elements Advanced Research (Nikon) was finally used to measure the fluorescence staining.
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