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C Peptide is part of the molecule of Proinsulin, that consists of three parts: C Peptide and two long strands of amino acids (called the alpha and beta chains) that later become linked together to form the insulin molecule. From every molecule of proinsulin, one molecule of insulin plus one molecule of C Peptide are produced. C peptide is released into the blood stream in equal amounts to insulin. A test of C peptide levels will show how much insulin the body is making. Insulin decreases blood glucose concentration. It increases cell permeability to monosaccharides, amino acids and fatty acids. It accelerates glycolysis, the pentose phosphate cycle, and glycogen synthesis in liver.
Our Abpromise guarantee covers the use of ab14181 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC||1/100 - 1/200.|
|WB||1/100. Can be blocked with Human C Peptide peptide (ab93903). The detection of a C-peptide in Western Blot is relatively difficult, because the peptide is only 5kDa big.|
|ICC/IF||Use at an assay dependent concentration.|
Formalin-fixed, paraffin-embedded human pancreas tissue stained for C Peptide with ab14181 (1/100 dilution) in immunohistochemical analysis. A Donkey anti-rabbit Alexa-Fluor® 488 conjugated secondary antibody was used at a 1/400 dilution.
ab14181 staining human normal pancreas. Staining is localized to the cytoplasm.
Left panel: with primary antibody diluted 1:2000.
Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with haematoxylin and coverslipped under DePeX.
Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.