Anti-SMARCC1 antibody (ab22355)

Western blot - SMARCC1 antibody (ab22355)

All lanes : Anti-SMARCC1 antibody (ab22355) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 ug
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 ug with SMARCC1 peptide (ab24351) at 1 µg/ml

Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

Predicted band size : 122 kDa
Observed band size : 150 kDa (why is the actual band size different from the predicted?)
Additional bands at : 100 kDa (possible cleavage fragment,cross reactivity),40 kDa (possible cleavage fragment,cross reactivity),85 kDa (possible cleavage fragment,cross reactivity).

ab22355 detects a band at 150 kDa which corresponds in size to that seen for SMARCC1, which was originally identified as a ~150 kDa polypeptide and termed BAF155. It also detects some smaller proteins at approximately 100, 85 and 40 kDa which may be degradation products. All of these bands are competed away by the addition of the immunizing peptide, suggesting that the interaction is specific.

Immunocytochemistry/ Immunofluorescence - SMARCC1 antibody (ab22355)

ICC/IF image of ab22355 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab22355, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - SMARCC1 antibody (ab22355)

Image courtesy of Human Protein Atlas

ab22355 staining SMARCC1in human kidney. Paraffin embedded human kidney tissue was incubated with ab22355 (1/2000 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab22355 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

Further results for this antibody can be found at www.proteinatlas.org