The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: Prediluted ready for use. Perform heat mediated antigen retrieval before commencing with IHC staining protocol by boiling tissue sections in 10mM citrate buffer pH 6.0 for 10-20 mins, followed by cooling at RT for 20 mins.
All five immunoglobulin classes share the same basic four polypeptide chain structure of two heavy-chains and two light chains. There are five heavy chain types, and two light-chain types (Kappa and Lambda) both having a molecular weight of 22.5kDa. Any heavy-chain type can associate with either light-chain type, but on any immunoglobulin molecule both light-chains are of the same type. Kappa and Lambda consist of a variable region and a constant region and can easily be differentiated by the antigenic properties of the constant region. The ratio of Kappa to Lambda is 70:30 , the vast majority of which is bound to heavy-chain in immunoglobulin. In normal individuals low levels of free light-chain arepresent in serum (kappa, 1.6-15.2 mg/L; Lambda, 0.4-4.2mg/L), with the occurrence of multiple myeloma or other B-cell malignancies these levels can be greatly elevated and can be found at high levels in the urine (Bence-Jones proteins).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lambda Light chain antibody, prediluted (ab74316)This image is courtesy of an anonymous Abreview
ab74316 staining Lambda Light chain in Rat lymph node tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 25°C; antigen retrieval was by heat mediation in EDTA pH9. Samples were incubated with primary antibody for 16 hours at 25°C. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.