In order to study the leukocytes types that could bind to IL-27β, head kidney leukocytes were incubated with 1 μg rOnIL-27β on ice for 30 min. After two times of wash with FACS buffer, mouse anti-His antibody (1:1000, BBI life sciences) and Alexa Fluor 647-conjugated goat anti-mouse IgG H&L (1:2000, Abcam) were sequentially incubated on ice away from light for another 30 min. After incubation, wash with FACS buffer twice. The same method was used for the final staining of the samples with anti-tilapia CD3ε monoclonal antibody. Another control group was not incubated with recombinant protein, but with secondary antibody to eliminate the possibility of non-specific binding. The stained cells were suspended in the flow tube with FACS buffer solution and analyzed by flow cytometry.
Mouse anti his antibody
Manufactured by BBI Lifesciences
The Mouse anti-His antibody is a laboratory reagent used to detect and purify recombinant proteins containing a histidine (His) tag. It specifically binds to the His tag, allowing for the identification and isolation of the target protein.
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Lab products found in correlation
2 protocols using mouse anti his antibody
1
Characterization of IL-27β-Binding Leukocytes
2
Identifying TGF-β1 Target Cells in Tilapia
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To investigate the potential target cells of TGF-β1, spleen lymphocytes were incubated with 1 μg recombinant His-tag TGF-β1 on ice for 30 min. After two times of wash with FACS buffer, 1:1000-diluted mouse anti-His antibody (BBI life sciences) and 1:2000-diluted Alexa Fluor 405–conjugated goat anti-mouse IgG H&L (Abcam) were successively added and incubated on ice for another 30 min. Finally, the samples were stained with anti-tilapia CD4-1 and anti-tilapia CD3ε mAbs as described above. The stained cells were resuspended with FACS buffer and analyzed by flow cytometry.
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