A2BP1 antibody (N-Term)

Catalog No. ABIN1951829

The Mouse Monoclonal anti-A2BP1 antibody has been validated for WB, ICC and IF. It is suitable to detect A2BP1 in samples from Human, Mouse, Rat and Cow.

Quick Overview for A2BP1 antibody (N-Term) (ABIN1951829)

Target: A2BP1 (Ataxin 2-Binding Protein 1 (A2BP1))

Reactivity: Human, Mouse, Rat, Cow

Host: Mouse

Clonality: Monoclonal

Conjugate: This A2BP1 antibody is un-conjugated

Application: Western Blotting (WB), Immunocytochemistry (ICC), Immunofluorescence (IF)

Product Details anti-A2BP1 Antibody

Binding Specificity: AA 1-100, N-Term

Specificity: Immunostaining Cell Cultures
1. Draw of culture medium with aspirator and add 1 ml of 3.7 % formalin in PBS solution to the dish. (make up from 10mls
Fisher 37% formalin plus 90mls PBS, the Fisher formalin contains 37% formaldehyde plus about 1% methanol which may be
relevant sometimes). Let sit at room temp for 1 minute. (can add 0.1% Tween 20 to PBS used here and all subsequent steps
to reduce background, probably best not to do this first time round though as it may extract your antigen or help wash your
cells off the dish).
2. Take off the formalin/PBS and add 1ml of cold methanol (-20°C, kept in well sealed bottle in fridge). Let sit for no more
than 1 minute.
3. Take off methanol and add 1ml of PBS, not letting the specimen dry out. To block nonspecific antibody binding can add
~10ml (=1%) of goat serum (Sigma), and can incubate for 30 minutes. Can then add antibody reagents. Typically 100ml of
hybridoma tissue culture supernatent or 1ml of mouse ascites fluid or crude serum. Incubate for 1 hour at room temp. (or
can go at 37°C for 30 minutes to 1 hour, or can do 4°C overnight, exact time not too critical). Can do very gentle shaking for
well adherent cell lines (3T3, Hek293 etc.).
4. Remove primary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give
three washes in PBS.
5. Add 0.5 mls of secondary antibody. These are fluorescently labeled Goat anti mouse antibodies and are conjugated to
ALEXA dyes and are from Molecular probes (Eugene Oregon, the ALEXA dyes are sulphonated rhodamine compounds and
are much more stable to UV than FITC, TRITC, Texas red etc.). Typically make 1:2,000 dilutions of these secondaries in
PBS plus 1% goat serum, BSA or non fat milk carrier. Incubate for 1 hour at room temp. (or can go at 37°C for 30 minutes to
1 hour, or can do 4°C overnight). Can do gentle shaking for well adherent cell lines (3T3, HEK293 etc.).
6. Remove secondary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give
three washes in PBS.
7. Drop on one drop of Fisher mounting medium onto dish and apply 22mm square coverslip. View in the microscope!




Immunostaining Tissue

Purification: Aff - Purified

Immunogen: Purified human recombinant protein fragment structure from E. Coli. corresponding to N terminal AA 1-100 of Human FOX1

Isotype: IgG1

Application Details

Application Notes: Immunofluorescence: 1:1:500-1:1,000 , Western Blot: 1:1,000-1:2,000, Dilutions listed as a recommendation. Optimal dilution should be determined by investigator.

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 1.0 mg/mL

Buffer: Purified tissue culture supernatant in PBS with 10mM sodium azide preservative.

Handling Advice: Avoid repeated freeze-thaw cycles.

Storage: 4 °C

Storage Comment: Antibody can also be aliquotted and stored frozen at -20° C to -70° C in a manual defrost , freezer for six months without detectable loss of activity. The antibody can be stored at 2° - 8°C for 1 month without detectable loss of activity.

Target Details for A2BP1

Target: A2BP1 (Ataxin 2-Binding Protein 1 (A2BP1))

Alternative Name: FOX1 / A2BP1