The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ELISA: Use at an assay dependent dilution.
Flow Cyt: Use at an assay dependent dilution.
ICC/IF: Use at an assay dependent dilution (PMID 19608744).
IP: Use at an assay dependent dilution.
WB: Use at an assay dependent dilution.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Immunocytochemistry/ Immunofluorescence - Blood Group AB antigen antibody [Z5H-2/Z2A] (ab24223)Image from Higgins MA et al, J Biol Chem. 2009 Sep 18;284(38):26161-73. Epub 2009 Jul 16, Fig 2. DOI 10.1074/jbc.M109.024067
ab24223 staining Blood Group AB antigen in A549 cells by Immunocytochemistry/ Immunofluorescence. Cells were rinsed in PBS and fixed in 4% paraformaldehyde in PBS (15 minutes), rinsed, and blocked in 5% lamb serum in PBS/Tween 20 (15 minutes, room temperature). Cells were rinsed three times in PBS before application of primary antibodies. ab24223 diluted 1/400 in 5% lamb serum in PBS and applied to the fixed cells overnight (4 °C). Specimens were rinsed three times in PBS, and Alexa 568-conjugated goat anti-mouse IgG secondary antibody diluted 1/800 in PBS was applied. Counterstained with DAPI.
Higgins MA et al. Differential recognition and hydrolysis of host carbohydrate antigens by Streptococcus pneumoniae family 98 glycoside hydrolases. J Biol Chem284:26161-73 (2009).
Read more (PubMed: 19608744) »