Determination of residual erythrocytes in rat tissue homogenates using commercially available anti-red blood cell sera.
A method to determine the residual erythrocyte content in rat tissue homogenates by means of radial immunodiffusion (Mancini et al., 1965) is described. The method makes use of commercially available antisera against rat erythrocytes. Tissues were homogenized after addition of digitonin and applied on Mancini plates. To measure within the linear part of the calibration curve, the tissue homogenates were diluted to concentrations between 0.062% and 2.5% wet weight content, depending on the type of tissue and on the degree of bleeding before the death of the animal. Recovery data of added blood as well as a comparison with results from organ distribution studies with 59Fe-labeled erythrocytes demonstrated that this simple method is sufficiently reliable and sensitive. The present procedure is to control experiments in which drug tissue levels (e.g., antibiotics) are to be determined and in which the drug content of residual blood is likely to bias the results to an unknown extent. It can be used as well in organ distribution studies of substances with a high affinity for erythrocytes (e.g., Pb; As in rats). It is possible to individually control the success of procedures without the use of isotopes--such as exsanguination or vascular perfusion--that are performed in order to reduce the blood content in the organs under consideration.
Schümann
K
K
Walther Straub-Institut of Pharmacology and Toxicology, Ludwig-Maximilians-Universität, München, FRG.
Kreppel
H
H
Elsenhans
B
B
D000818:Animals; D002458:Cell Fractionation; D004072:Digitonin; D004912:Erythrocytes / Q000276:immunology; D005779:Immunodiffusion; D007413:Intestinal Mucosa / Q000473:pathology*; D007583:Jejunum; D007668:Kidney / Q000473:pathology*; D008099:Liver / Q000473:pathology*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains
D004072:Digitonin