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Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human CD3.
(Peptide available as ab16206.)
Our Abpromise guarantee covers the use of ab16044 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 5 µg/ml.|
|IHC-P||Use at an assay dependent concentration.|
|WB||Use a concentration of 0.5 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).|
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab16044 overnight at 4°C. Antibody binding was detected using an anti-rabbit IgG VHH single domain antibody conjugated to HRP, and visualised using ECL development solution ab133406.
ab16044 staining CD3 in human thymus tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in buffer and blocking (5 minutes/peroxidase block then 10 minutes/protein block) for 45 minutes at 20°C.The primary antibody was diluted, 1/250 and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.
Rabbit polyclonal antibody (ab16044) detecting CD3 protein on PFA perfused Rat spleen section. The section was blocked with 10% donkey serum in PBS for 1hour at 24°C and then incubated with a 1/200 dilution of ab16044 for 24 hours at 4°C. Secondary antibody is a donkey anti-rabbit Alexa 568 (1/1000) and nuclei stained using the Hoechst method are depicted in blue . CD3 expressed in T cells but not in the B-cell germinal centre.