Materials and reagents Expand Equipment Thermo Fisher Scientific round (12 mm diameter) cover glass (thickness No 1.5). Thermo Fisher Scientific ColorFrostTM Plus slides made of positively charge uncoated glass. Blocking buffer Expand BLOCK Solution NaCl 8 g KCl 0.2 g Na2HPO4 1.44 g KH2PO4 0.24 g Tween-20 2 ml Fetal bovine serum 50 ml […]
Differentiating principal cell from intercalated cell Expand The collecting duct in the kidney consists in three types of cells, principal cells, alpha-intercalated cells and beta-intercalated cells. The principal cells are primarily responsible for salt and water transport while alpha- and beta-intercalated cells are responsible for acid-base transport. Figure legend Anti-Aqp2 antibody labels the luminal membrane […]
The skin Expand The skin, or integument, consists of two major layers: the epidermis, composed of stratified squamous epithelium that is keratinized, and the dermis, composed of connective tissue. Figure legend E-cadherin proteins label the epithelial cell junctions in the epidermis of the skin from a mouse embryo at E12.5 on FFPE section. The kidney […]
Citrate buffer at pH 6.0 Expand FFPE sections were obtained from formalin (10% neutral formaldehyde) fixed mouse kidney tissues. Antigens on FFPE sections were retrieved by citrate solution at pH 6.0. Two BiCell Scientific antibodies, raised in rat and rabbit respectively, were incubated with FFPE sections to detect two independent antigen proteins. Figure legend Fluorescent […]
Low-magnification fluorescent imaging Expand FFPE sections were obtained from formalin (10% neutral formaldehyde) fixed mouse kidney tissues. Antigens on FFPE sections were retrieved by citrate solution at pH 6.0. Two BiCell Scientific antibodies, raised in rat and rabbit respectively, were incubated with FFPE sections to detect two independent antigen proteins. Figure legend Fluorescent IHC on […]
IHC on cryosection Expand Cryosections are obtained from lightly fixed (<4% PFA), and rapidly frozen tissues, which offers a key advantage of antigen preservation in situ in tissue sections. Tissue morphology is not as well preserved as FFPE sections. Figure legend Cryosection imaging shows the Mucin-1 proteins in the luminal membrane of the epithelium lining […]
Materials and Reagents Expand Equipment Leica CM1950 automatic sectioning cryostat. Thermo Fisher Scientific round (12 mm diameter) cover glass (thickness No 1.5). Thermo Fisher Scientific ColorFrostTM Plus slides made of positively charge uncoated glass. Tissue-tek Expand Tissue-tek OCT 4583 compound from VWR. Tissue-tek cryomolds from VWR. Fixation Buffers Expand 2% paraformaldehyde in PBS NaCl […]
The cerebral cortex Expand The mitochondrion shows a pattern of punctate appearance in the cytoplasm of the cell body in the neurons of the cerebral cortex. Figure legend TOMM20 antibody labels the mitochondria in the pyramidal neurons of the cerebral cortex in the mouse brain. The hippocampus Expand The mitochondrion shows a pattern of punctate […]
Preparation of Myeloma Cells Expand Prewarm all culture media to 37oC. Thaw the parental myeloma cells (Sp2/0-Ag14 or X63-Ag8.653) from liquid nitrogen and culture in BiCell-mAb Medium MY for 3-5 days to fully recover cell growth to reach 1 x 106/ml cell density. Split cells at 1:5 into fresh BiCell-mAb Medium MY and let grow […]
The skin Expand The ER in the epidermal cells shows a pattern of perinuclear network that is continuous with the nuclear membrane. Figure legend Calreticulin antibody labels the ER in the epithelium of the skin from a mouse embryo at E12.5. The liver Expand The ER in the hepatocytes shows a pattern of perinuclear network […]
"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."
Jerrold R. Turner, M.D., Ph.D.
"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."
Univ.-Prof. Jay Gopalakrishnan PhD
"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."
Julie Craft Van De Weghe, PhD
"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"
Divyesh Joshi, PhD
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