Anti-MAGEA1 抗体 [SP188] (ab227664)

IHC image of Magea1 staining in a section of formalin-fixed paraffin-embedded human normal testis* performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab227664, 0.05ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Negative control image: IHC image of MAGEA1 staining in a section of  formalin-fixed paraffin-embedded human normal skeletal muscle* performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab227664, 0.05ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Flow cytometry overlay histogram showing left A-431 positive cells and right negative MCF7 stained with ab227664 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab227664) (1x 10⁶ in 100μl at 1.0μg/ml (1/2100)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Flow Cytometry analysis of SK-OV-3 (Human ovarian cancer epithelial cell) cells labeling MAGEA1 with purifiedab227664 at 1:20 dilution (7.25 µg/ml) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor© 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

Flow cytometric analysis of A431 (human epidermoid carcinoma cell line) cell line labeling MAGEA1 with ab227664 at 1/400 dilution (green) compared with a Rabbit IgG negative control (blue).

Formalin-fixed, paraffin-embedded human skin squamous cell carcinoma tissue stained for MAGEA1 using ab227664 at 1/100 dilution in immunohistochemical analysis.