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Full length protein corresponding to Mouse IgG1.
Our Abpromise guarantee covers the use of ab193651 in the following tested applications.
|WB||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|IHC||Use at an assay dependent concentration.|
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 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 ab8226 overnight at 4°C. Antibody binding was detected using an anti-mouse IgG1 VHH single domain antibody conjugated to HRP (ab193651), and visualised using ECL development solution ab133406.
IHC image of Anti-Mouse IgG1 VHH Single Domain Antibody (HRP) (isotype specific) (ab193651) staining in formalin fixed paraffin embedded normal human colon tissue section.
The section was dewaxed and then pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with mouse IgG1 monoclonal antibody to Histone H1.0 (ab11080, 2.5µg/ml) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. Endogenous peroxidases were quenched using 1.6% (v/v) hydrogen peroxide in TBS containing 0.025% (v/v) Triton X-100 for 30 minutes at room temperature, with agitation. The secondary antibody, Anti-Mouse IgG1 VHH Single Domain Antibody (HRP) (isotype specific) (ab193651, 0.05µg/ml) was then applied for 1 hour at room temperature in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA before being developed for 10 minutes at room temperature using Steady DAB/Plus (ab103723). The section was then counterstained with hematoxylin and mounted with DPX.
The negative control (secondary antibody only, no primary) inset shows no staining, demonstrating secondary antibody specificity.
For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.