Anti-Geminin antibody (ab12147)

Western blot

All lanes : Anti-Geminin antibody (ab12147) at 1/500 dilution

Lane 1 : 293 lysate
Lane 2 : 293 lysate with Geminin peptide (ab12408) at 1 µg/ml

Lysates/proteins at 5 µg/ml per lane.

developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 23.5 kDa
Observed band size : 33 kDa (why is the actual band size different from the predicted?)


The lower band represents Geminin (33 kDa - predicted molecular weight 23 kDa), and is blocked by the immunising peptide, the upper band is likely to be non-specific as it is not blocked by the peptide.

Immunocytochemistry/ Immunofluorescence - Geminin antibody (ab12147)

ab12147 (1/400) staining geminin in assynchronous HeLa cells. Cells were fixed in paraformaldehyde and counterstained with DAPI in order to highlight the nucleus. Secondary antibody: goat anti-rabbit IgG conjugated to Cy3. Please refer to abreview for further details.

This image is courtesy of an Abreview submitted by Dr Kirk McManus

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Geminin antibody (ab12147)

Image courtesy of Human Protein Atlas

ab12147 staining in human testis, showing staining of the cells in the ductus seminiferus (in brown). Paraffin embedded testis tissue was incubated with ab12147 (1:175 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab12147 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

Western blot - Geminin antibody (ab12147)

Lane 1 : Anti-Geminin antibody (ab12147) at 1 µg/ml
Lane 2 : Anti-6X His tag® antibody - ChIP Grade (ab9108) at 1 µg/ml

Lane 1 : Geminin protein (His tag) (ab40570)
Lane 2 : Geminin protein (His tag) (ab40570)

Lysates/proteins at 0.1 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 23.5 kDa
Observed band size : 32 kDa (why is the actual band size different from the predicted?)

Western blot - Geminin antibody (ab12147)

Anti-Geminin antibody (ab12147) at 1/500 dilution + Cell lysate prepared from human HeLa nuclear extract at 50 µg

Secondary
HRP-conjugated goat poyclonal to rabbit IgG at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 23.5 kDa
Observed band size : 33 kDa (why is the actual band size different from the predicted?)
Additional bands at : 95 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 10 minutes

This image is a courtesy of Anonymous Abreview