Doublecortin Antikörper

Produktnummer ABIN1951823

Der Maus Monoklonal Anti-Doublecortin-Antikörper wurde für WB, IF und ICC validiert. Er ist geeignet, Doublecortin in Proben von Human, Maus, Ratte, Rind (Kuh), Schwein, Pferd, Huhn und Primate zu detektieren.

Kurzübersicht für Doublecortin Antikörper (ABIN1951823)

Target: Doublecortin (DCX)

Reaktivität: Human, Maus, Ratte, Rind (Kuh), Schwein, Pferd, Huhn, Primate

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser Doublecortin Antikörper ist unkonjugiert

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

Produktdetails anti-Doublecortin Antikörper

Spezifität: 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 or rabbit 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
Solutions
PBS - sodium phosphate-buffered (100 mM, pH 7.2) isotonic (0.9% NaCl, w/v) saline Antibody dilution buffer (PBS with
0.1% non-ionic detergent, such as Triton X-100 or Tween-20) fluorescein anti-fading reagent -- Make up a 2 mg/mL
phenylene diamine solution in PBS (phenylene diamine requires extensive vortexing to put it into solution). Once the

Aufreinigung: Purified

Immunogen: Recombinant GFAP expressed in bacteria and highly purified. Subsequent boosts were performed with GFAP purified from a Triton X-100 extract of myelin associated material from bovine spinal cord.

Isotyp: IgG1

Anwendungsinformationen

Applikationshinweise: Immunofluorescence: 1:1,000 , Immunohistochemistry: 1:1,000 , Western Blot: 1:5,000. Dilutions listed as a recommendation. Optimal dilution should be determined by investigator.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Liquid

Buffer: Liquid with 10mM Sodium Azide

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.

Handhabung: Avoid repeated freeze-thaw cycles.

Lagerung: 4 °C

Informationen zur Lagerung: Antibody can 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.

Details zu Doublecortin

Target: Doublecortin (DCX)

Andere Bezeichnung: Doublecortin