Immunohistochemical analysis of paraffin-embedded human appendix tissue using BD-PB3883 performed on the Leica® BOND RXm. Tissue was fixed with formaldehyde at room temperature; antigen retrieval was by heat mediation with a EDTA buffer (pH9. 0). Samples were incubated with primary antibody (1:1000) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.

Immunofluorescent analysis of U251 cells, using KIT Antibody conjugated Phalloidin (red).

KIT Antibodywestern blot analysis in A549 cell line lysates (35μg/lane).This demonstrates the KIT antibody detected the KIT protein (arrow).

Fluorescent confocal image of an SY5Y cell stained with BD-PB3883 KIT antibody. SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min), then incubated with BD-PB3883 KIT primary antibody (1:500, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-mouse antibody (green) was used (1:1000, 1h). Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (5.25 μM, 25 min). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 3 min). Note the highly specific localization of the KIT immunosignal to the membranes, especially the plasma membrane. ibid. References for SY5Y : 1. Ross RA, et al. Coordinate morphological and biochemical interconversion of human neuroblastoma cells. J. Natl. Cancer Inst. 71: 741-749, 1983. [PubMed: 6137586]; 2. Biedler JL, et al. Multiple neurotransmitter synthesis by human neuroblastoma cell lines and clones. Cancer Res. 38: 3751-3757, 1978. [PubMed: 29704]. Ibid.