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Query: UNIPROT:P30536 (PBS)
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Sensitive identification of human blood and the determination of ABO blood group from a minute bloodstain were simultaneously carried out by a direct ELISA-ABC method. A cotton thread (1 cm in length) stained with 1 microliter of human or animal blood was stored for 2-4 weeks at room temperature. Hemoglobin (Hb) of the bloodstained thread was gently extracted with 100 microliters of PBS at room temperature, and the thread was washed with PBS to dehemoglobinize. And ABH blood group antigens were extracted from the same dehemoglobinized thread with 100 microliters of 5% ammonia solution at 56 degrees C. The extracts of PBS and ammonia were two-fold serially diluted with 0.1 M sodium carbonate buffer, coated to the wells of a flat bottomed microplate. The PBS extract was tested with a biotinylated antibody against human HbA0 for identification of human blood. Human blood was clearly distinguishable from bloods of other species including Japanese monkey. The minimum detection limit of human blood of the PBS extract of the bloodstained thread was 1:40,960 (3.4 ng Hb), and the limit was found to be approximately 200 times higher than that obtained by a leucomalachite green test or by a precipitation ring test using anti-human HbA serum. The ammonia extract was tested with biotinylated anti-A, anti-B and anti-H antibodies for ABO blood grouping. ABH antigens of the ammonia extract of the bloodstained thread were clearly detected. The minimum determination limits of blood group A, B, AB and O of the ammonia extracts were 1:160, 1:160, 1:80 and 1:160, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Sensitive identification of human blood and simultaneous determination of ABO blood group from a minute bloodstain by an ELISA-ABC method]. 130 54

A suspension of washed human erythrocytes (2%) in PBS was mixed with an equal volume of 1 mM glutaraldehyde (GA) and allowed to stand at laboratory temperatures, followed by washing with normal saline. The agglutinability of the erythrocytes toward anti-A, anti-B, anti-M and anti-N reagents remained unchanged after GA treatment shorter than 20 minutes, and the agglutinability toward anti-C, anti-c, anti-D, anti-E, anti-e, anti-Lea, anti-Leb and anti-P1 did not decrease after treatment for 10 minutes. GA treatment for longer periods of time than the above caused a decrease of the reactivities. The agglutinability toward an anti-H (Ulex europaeus) and other lectins increased after 10 to 30 minutes of GA treatment and decreased after 40 minutes or more of exposure to GA. These results indicate that the immunologic agglutinability of erythrocytes were practically unchanged after a 10 minutes treatment with 1 mM GA (a mild fixation procedure hereafter called "partial fixation"). The properties of the partially fixed erythrocytes were closely similar to those of untreated erythrocytes as regards cell volume, membrane fragility in hypotonic solutions (measured by a modification of the Ribiere method), and membrane fragility under continuous shaking. The resistance of the partially fixed erythrocytes heating up to 50 degrees C was superior to that of the untreated erythrocytes. From these results, the deformability of the partially fixed erythrocytes was concluded to be similar to that of untreated erythrocytes. After storage for 6 months at 4 degrees C in Alsever solution containing adenine and inosine, the shapes of the untreated erythrocytes changed to "spherocytes" or "echinocytes", whereas the partially fixed cells retained the original discocyte shape. Blood grouping laboratory tests were performed with the partially fixed erythrocytes as indicators. Both anti-A and anti-B agglutinins in normal human sera could be detected without difficulty. Presence of irregular agglutinins, such as anti-H and anti-N, in healthy donors' sera could also be detected, as with the freshly obtained erythrocytes. The detection limits of an incomplete anti-D agglutinin were equal to those in the tests with untreated erythrocytes. The partially fixed erythrocyte (stored at 4 degrees C for 2.5 to 3 months) were used as indicators of ABO-blood grouping from blood stains, saliva stains and hairs by the agglutinin-inhibition test, absorption-elution test or mixed agglutination test. The results obtained were practically equal to those of the tests with freshly obtained erythrocytes, indicating the availabilities of the partially fixed erythrocytes.
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PMID:[Preparation of partially fixed erythrocytes and its application for immunohematologic examination]. 211 66

Twenty-five permanent teeth, including eight carious ones whose pulp cavities had been exposed, were used for this research 3-5 weeks after extraction. Phosphate-buffered saline (PBS, at pH 7.2) was employed to extract ABO blood group substance from tooth powder. ABO grouping was performed on blood-stained compresses from the extraction wound (as controls), tooth fragment, tooth powder, and cotton fibers immersed in PBS extract by absorption-elution (AE) technique and on the PBS extracts by the two-dimensional absorption-inhibition (2-D AI) technique. It was found that blood grouping in PBS extracts by 2-D AI yielded reliable results: no false positive results, and a high rate of correct grouping, (24/25), while blood grouping on other dental materials, such as tooth fragments, tooth powders, immersed fibers, by AE gave an unacceptable rate of false positive/negative results.
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PMID:ABO blood grouping on dental tissue. 835 10

A simple immunochromatographic method was used to determine ABO blood groups and secretor status from saliva stains. Saliva specimens were serially diluted with PBS and dotted onto nitrocellulose strips (10 mm x 70-100 mm). One end of the strips was dipped in two drops of 0.1% orange-red gold particles (GP) which were coated with an anti-A, anti-B or anti-H blood group reagent. The GP migrated along the surface of the strips producing red spots where positive antigen-antibody reactions occurred. In non-secretors, the end-points of saliva dilution were lower than in secretors. The results of this test were obtained within 2 to 3 minutes. Moreover, the GP reagents utilized were quite inexpensive. The principles underlying this method might be useful in species and organ identification.
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PMID:[Simple determination methods for blood grouping from biological stains. 2. Immunochromatographic determination of ABO blood groups from saliva specimens]. 978 Jun 63