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.
IHC-Fr: Use at an assay dependent dilution.
IHC-P: Use at a concentration of 1 µg/ml.
IP: Use at an assay dependent dilution.
RIA: Use at an assay dependent dilution.
WB: Use at an assay dependent dilution. Predicted molecular weight: 66 kDa.
ICC/IF: Use at an assay dependent dilution From (PubMed:17353362)
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
ICAM proteins are ligands for the leukocyte adhesion protein LFA-1 (integrin alpha-L/beta-2). ICAM3 is also a ligand for integrin alpha-D/beta-2.
Belongs to the immunoglobulin superfamily. ICAM family. Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
Upon stimulation by a physiologic stimuli becomes rapidly and transiently phosphorylated on serine residues. N-glycosylated; glycans consist of a mixture of tri- and tetra-antennary complex-type chains and high-mannose chains.
Ab10804 staining Human malignant stomach. Staining is localised to the membrane. Left panel: with primary antibody at 1 ug/ml. 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 buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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.
Immunocytochemistry/ Immunofluorescence - Anti-ICAM3 antibody [ICAM3.1] (ab10804)Image from Heasman SJ et al, J Cell Biol. 2010 Aug 23;190(4):553-63, Fig 5. doi/10.1083/jcb.201002067
ab10804 staining ICAM3 in Y27632-treated CEM T cells by Immunocytochemistry/ Immunofluorescence. CEM T cells were fixed for 15 minutes with 4% paraformaldehyde. Cells were permeabilized with 0.1% Triton X-100, blocked
with 1% BSA, and incubated with ab10804 (1/50), for 1 hour
followed by Alexa Fluor 488–conjugated anti–mouse
IgG secondary antibodies (1/200).
Heasman SJ et al. Coordinated RhoA signaling at the leading edge and uropod is required for T cell transendothelial migration. J Cell Biol190:553-63 (2010).
Read more (PubMed: 20733052) »