anti-Actin mouse monoclonal, 2G2, supernatant
- Mouse monoclonal
- Suitable for ICC/IF, IHC and WB
- Reacts with bovine, chicken, human, plant and rabbit
- Isotype: IgM
|Application||ICC, IHC, WB|
|Purification||Hybridoma cell culture supernatant|
|Reactivity||Bovine, Chicken, Human, Plant, Rabbit|
|Intended use||Research use only|
|Immunogen||Profilin actin complex from calf thymus|
|Formulation||Contains 0.09% sodium azide|
|Tested applications||Tested dilutions|
|Immunocytochemistry (ICC)/ Immunofluorescence (IF)||Assay dependent (IF only after methanol fixation)|
|Immunohistochemistry (IHC) - frozen||Ready-to-use|
|Immunohistochemistry (IHC) - paraffin||Ready-to-use (protease treatment recommended)|
|Western Blot (WB)||Assay dependent|
Specific for skeletal and non-muscle actin isoforms (43 kDa). Epitope location: NVPAMYVAVLDSGVTHNVPIYHAIMRLDLA.
When using antibody 2G2 in immunolocalization studies the presence of nuclear actin could be demonstrated e.g. in cultured cells after fixation with 4% paraformaldehyde. Immunostaining with cytoplasmic actin was only positive after fixation with methanol (Gonsior et al., 1999).
Reactivity on cultured cell lines: PtK2, SR-NRK, NRK-49F, L6 cells, rat fibroblasts, NIH-3T3, mouse myoblast and myotube cells.
Gonsior SM, Platz S, Buchmeier S, Scheer U, Jockusch BM, Hinssen H: Conformational difference between nuclear and cytoplasmic actin as detected by a monoclonal antibody. J Cell Sci 112, 797-809 (1999)
Q & A's
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The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at firstname.lastname@example.org.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
- Supernatant and supernatant concentrate: This format contains hybridoma cell culture supernatant. The antibody is not purified and the antibody concentration is not determined. The antibody concentration may vary from lot to lot. Therefore we recommend to titrate the optimal concentration for the application used for each new lot.
- Lyophilized, purified: This format contains purified antibody in lyophilized form. The reconstitution of this antibody is described in the datasheet. The buffer composition after reconstitution is also mentioned on the datasheet.
- Liquid, purified: This format contains purified antibody in liquid format. The concentration is mentioned on the datasheet.
- Prediluted, purified: This format contains purified antibody in liquid format. Most antibodies in this format are diluted to be ready-to-use for IHC with standard tissue. But some antibodies of this format need further dilution for IHC. This is mentioned on the datasheet.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
- Drain off medium from cells grown on cover slips.
- Permeabilize cells with solution I supplemented with 0.2% Triton X-100 (solution I: PBS plus 5.5 mM glucose, 2 mM Mg-chloride, 2 mM EGTA, 5 mM Pipes; pH 6.1), immerse for 90 sec at RT.
- Fixation of cells with 4% paraformaldehyde in solution I for 5 min at RT.
- Wash cells 3 x with TBS.
- Block with the serum of the species in which the secondary antibody was raised for 30 min at RT.
- Incubate with 2G2 antibody 1 h at RT.
- Wash 3 x with TBS.
- Incubate with secondary antibody for 30 min at RT.
- Wash 3 x with TBS.
- Remove excess buffer.
- Mount with Mowiol and cover with a glass slide (or a second cover slip).
- Examine under the microscope.