anti mouse cd117 mabs  (Bio X Cell)

Journal: Frontiers in Microbiology

Article Title: Development and characterization of monoclonal antibodies against the extracellular domain of African swine fever virus structural protein, CD2v

doi: 10.3389/fmicb.2022.1056117

Figure Lengend Snippet: Characterization and identification of mAbs. (A) Determination of the serum titer of the mice before hyperimmunization by indirect ELISA. (B) Determination of the mAb titers. A P/G value greater than or equal to 2.1 is considered positive, and the corresponding maximum dilution is the titer of serum or mAbs. (C) Four mAbs were identified by IFA. The CD2v recombinant protein gene was linked to a vector containing the EGFP label and was transfected into 293 T cells. A monoclonal antibody was used as the primary antibody, and a goat anti-mouse Alexa Fluor 647 fluorescent antibody was used as the secondary antibody to display red fluorescence. DAPI was used as the nuclear staining fluid. The control group was used as a negative control. (D) Four mAbs were identified with CD2v recombinant protein by a western blot assay. M represents the protein marker, monoclonal antibody 11E2 was incubated in Lane 1, Lane 2 is 7A9, Lane 3 is 4C1 and Lane 4 is 2H10. (E) The reactivity of the mAbs with ASFV HLJ/18 strain. Scale bars, 200 μm. (F) SDS–PAGE analysis of purified monoclonal antibody 11E2. (G) CD2v mAbs evaluation by the blocking enzyme-linked immunosorbent assay. The African swine fever positive serum was serially diluted for 1:10 ( n = 20).

Article Snippet: The mAb subtypes were determined by a mouse mAb subtype identification kit (Proteintech, Wuhan, China).

Techniques: Indirect ELISA, Recombinant, Plasmid Preparation, Transfection, Fluorescence, Staining, Control, Negative Control, Western Blot, Marker, Incubation, SDS Page, Purification, Blocking Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Innate Immunity

Article Title: Calcineurin Aα Contributes to IgE-Dependent Mast-Cell Mediator Secretion in Allergic Inflammation

doi: 10.1159/000520040

Figure Lengend Snippet: LMCs from CnAα -deficient mice have similar morphology and development compared to WT in vitro. a Representative genotyping results of WT, CnAα -deficient (KO), and CnAα heterozygous (HZ) samples with labeled DNA ladder. The predicted sizes of CnAα WT and CnAα mutant (neomycin cassette insertion) are 247 bp and 360 bp, respectively. b Representative image using toluidine blue staining of each genotype (original magnification ×100). c , d Flow cytometric detection of surface molecules of WT (left) and CnAα-deficient LMCs (KO, right) sensitized with anti-TNP IgE and then stained with either FITC-conjugated anti-CD117 ( c ) or anti-IgE antibodies ( d ). Red peaks are isotype controls. e Intracellular calcium flux was compared between FcεRI-mediated activated WT and CnAα -deficient LMCs. BP, base pairs; WT, wild-type; LMC, liver-derived mast cells; CnAα, calcineurin subunit A; FITC, fluorescein isothiocyanate; TNP, trinitrophenol; FcεRI, high-affinity receptor for immunoglobulin E; IgE, immunoglobulin E.

Article Snippet: FITC-conjugated rat anti-mouse CD117 (clone: 2B8) mAb and isotype control FITC-rat IgG2b (eB149/10H5) were purchased from eBioscience (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: In Vitro, Labeling, Mutagenesis, Staining, Derivative Assay