UCHL1 anticorps

N° du produit ABIN1951827

Détail du produit anti-UCHL1 anticorps

Antigène: UCHL1 (Ubiquitin Carboxyl-terminal Esterase L1 (Ubiquitin Thiolesterase) (UCHL1))

Reactivité: Humain, Rat, Souris, Boeuf (Vache), Primate

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp UCHL1 est non-conjugé

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

Specificité: 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). For anti-fading, use Neuromics' i-BRITE Plus-Catalog#:
SF40000 or make your won 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 phenylene diamine is completely dissolved,
add an equal volume of glycerol and mix. This reagent will last about a week at -20
O
C. Discard this reagent when it starts to
turn dark brown.

Purification: Purified

Immunogène: Recombinant full length human PGP9.5 or UCHL1 purified from E. coli.

Isotype: IgG

Information d'application

Indications d'application: Immunofluorescence: 1:500 , Western Blot:1:2,000, Dilutions listed as a recommendation. Optimal dilution should be determined by investigator.

Restrictions: For Research Use only

Stockage

Format: Liquid

Buffer: Liquid with 10mM , sodium azide as a preservative.

Conseil sur la manipulation: Avoid repeated freeze-thaw cycles.

Stock: 4 °C

Stockage commentaire: 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.

Détails sur UCHL1

Antigène: UCHL1 (Ubiquitin Carboxyl-terminal Esterase L1 (Ubiquitin Thiolesterase) (UCHL1))

Autre désignation: UCHL1 / PGP9.5 (UCHL1 Produits)

Synonymes: anticorps PARK5, anticorps PGP 9.5, anticorps PGP9.5, anticorps PGP95, anticorps Uch-L1, anticorps UCH-L1, anticorps UCHL1, anticorps cb358, anticorps wu:fc55h08, anticorps park5, anticorps pgp9.5, anticorps uch-l1, anticorps MGC132191, anticorps uchl1, anticorps AW822034, anticorps C88048, anticorps R75593, anticorps UCHL-1, anticorps gad, anticorps ubiquitin C-terminal hydrolase L1, anticorps ubiquitin carboxyl-terminal esterase L1 (ubiquitin thiolesterase), anticorps ubiquitin C-terminal hydrolase L1 L homeolog, anticorps ubiquitin carboxyl-terminal esterase L1, anticorps ubiquitin carboxy-terminal hydrolase L1, anticorps UCHL1, anticorps uchl1, anticorps uchl1.L, anticorps LOC100304750, anticorps Uchl1

Pathways: Feeding Behaviour