For T and B cell phenotyping, cells isolated from blood, lamina propria, and caecal tonsils were stained with antibodies for CD3 (T cells), CD4 (T helper cells), and CD8 (T cytotoxic cells) (mouse anti-chicken CD3: PE; mouse anti-chicken CD4: FITC; mouse anti-chicken CD8: Cy5) according to methods described by Liermann et al. (5 (link)). Furthermore, additional samples of PBL's and cells isolated from lamina propria were incubated with antibodies for Bu1 (B cells) (mouse anti-chicken Bu-1: FITC; Southern Biotech; Birmingham, USA). For corresponding isotype controls either mouse IgG1 negative control: PE; mouse IgG1 negative control: FITC, or mouse IgG1 negative control: Cy5 (Southern Biotech; Birmingham, USA) were used. T and B cell subsets were measured by FACS Canto II (BD Bioscience, San Jose, USA). Therefore, at least 10,000 cells were considered. The evaluation of results and the compensation of non-specific signals indicated by the isotype controls were conducted by using the BD FACSDiva™ Software (BD Biosciences, San Jose, USA).
Mouse anti chicken bu 1
Manufactured by Southern Biotech
Sourced in United States
The Mouse anti-Chicken Bu-1 is a laboratory reagent used for the detection and identification of the Bu-1 antigen, which is expressed on chicken B cells. It can be used in various immunological techniques, such as flow cytometry and immunohistochemistry, to study B cell development and function in chicken models.
Automatically generated - may contain errors
2 protocols using mouse anti chicken bu 1
1
Phenotyping T and B Cells
2
Phenotypic Analysis of Chicken Lymphocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Isolated PBLs and cells isolated from spleen and CT were incubated with mAbs
against CD3 (T cells) (mouse anti-chicken CD3: PE; Southern Biotech; Birmingham,
AL), CD4 (T helper cells) (mouse anti-chicken CD4: FITC; Southern Biotech) and
CD8 (T cytotoxic cells) (mouse anti-chicken CD8: Cy5; Southern Biotech) for 30
min at room temperature in the dark. Further samples were incubated with an Ab
against Bu1 (B cells) (mouse anti-chicken Bu-1: FITC; Southern Biotech) under
conditions similar to those described above. Samples intended for corresponding
isotype controls were incubated with either mouse IgG1 negative control: PE;
mouse IgG1 negative control: FITC or mouse IgG1 negative control: Cy5 (Southern
Biotech). Thereafter, 1000 µl HEPES-buffered saline was added and the samples
were centrifuged at 250 g for 5 min at 4°C. Subsequently, the T
and B cell subsets were measured by FACS Canto II (BD Bioscience, San Jose, CA).
At least 10,000 cells were counted. The BD FACSDiva™ Software (BD Biosciences)
was used to evaluate results and compensate non-specific signals indicated by
isotype controls.
against CD3 (T cells) (mouse anti-chicken CD3: PE; Southern Biotech; Birmingham,
AL), CD4 (T helper cells) (mouse anti-chicken CD4: FITC; Southern Biotech) and
CD8 (T cytotoxic cells) (mouse anti-chicken CD8: Cy5; Southern Biotech) for 30
min at room temperature in the dark. Further samples were incubated with an Ab
against Bu1 (B cells) (mouse anti-chicken Bu-1: FITC; Southern Biotech) under
conditions similar to those described above. Samples intended for corresponding
isotype controls were incubated with either mouse IgG1 negative control: PE;
mouse IgG1 negative control: FITC or mouse IgG1 negative control: Cy5 (Southern
Biotech). Thereafter, 1000 µl HEPES-buffered saline was added and the samples
were centrifuged at 250 g for 5 min at 4°C. Subsequently, the T
and B cell subsets were measured by FACS Canto II (BD Bioscience, San Jose, CA).
At least 10,000 cells were counted. The BD FACSDiva™ Software (BD Biosciences)
was used to evaluate results and compensate non-specific signals indicated by
isotype controls.
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