Target cells were adjusted to a density of 1.0 × 106/mL, and 100 μL of cell suspension was transferred into flow tubes. The primary antibodies used were as follows: mouse anti-rat CD11b (BD, New Jersey, USA) and CD68 (Santa Cruz, Dallas, Texas, USA), and mouse IgA (BD) and IgG2b (BD), and were used for the identification of M0 macrophages; mouse anti-rat CD29 (eBioscience, San Diego, California, USA), CD90 (eBioscience), CD44 (Santa Cruz), CD79a (Santa Cruz), CD45 (eBioscience), and CD11b (BD) were used for the identification of PBMSCs; and mouse anti-rat CD206 (Santa Cruz) and CD86 (Santa Cruz), and mouse IgG1 (BD) and IgG2b (BD) were used for the identification of M1 and M2 macrophages. After 30 min of incubation with primary antibodies, cells were subsequently incubated with Alexa Fluor-conjugated rabbit or goat anti-mouse IgG secondary antibodies. After washing with PBS and fixing with 200 μL 4% paraformaldehyde, cell samples were analyzed by FACSCalibur flow cytometry (Becton Dickinson, USA), and data were analyzed using Cell Quest software.
Mouse iga
Manufactured by BD
Sourced in United States
Mouse IgA is a laboratory product used in various research applications. It is a type of immunoglobulin, specifically the IgA class, derived from mouse samples. This product can be utilized in experimental protocols that require the detection or measurement of mouse IgA.
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Lab products found in correlation
2 protocols using mouse iga
1
Phenotypic Characterization of Macrophages and PBMSCs
2
Investigating Intestinal IgA Transport
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On day three, the Transwells were removed from the various treatment conditions, and switched to base media (Advanced DMEM/F12 supplemented with 2mM L-glutamine only; no FBS, no antibiotics). 600μl of base media containing 3μg of mouse IgA (BD Pharmingen) was added to the lower compartment (final concentration of IgA = 5μg ml−1). In some experiments, 600μl of base media alone (no IgA) was added to the lower compartment. 100 μl of base media (with or without various treatments) was added to the upper compartment. Treatments included 1/10 fecal bacterial suspensions (bacterial pellet or supernatant fractions), 1/10 overnight anaerobic chopped meat bacterial cultures (live or freeze/thawed), and 1X Roche cOmplete protease inhibitor cocktail (Roche). The apical supernatant or cells were collected at 3 or 6 hours to evaluate the amount of pIgR/SC by immunoblotting or IgA by ELISA (Immunology Consultants Labs). Each experiment reflects unique biological replicates.
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