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Cytochrome P450, Family 19, Subfamily A, Polypeptide 1 (CYP19A1) (AA 400-502), (C-Term) antibody

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C-Term, AA 400-502
(22), (21), (13), (10), (6), (5), (4), (4), (2), (2), (2), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1)
Cow (Bovine), Horse (Equine), Human, Monkey, Rabbit
(135), (55), (38), (19), (9), (8), (6), (6), (3), (3), (3), (3), (3), (1), (1), (1), (1), (1), (1)
(92), (29), (20)
(5), (4), (4), (3), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1)
Western Blotting (WB), Immunocytochemistry (ICC), Immunofluorescence (IF)
(109), (45), (38), (37), (23), (16), (5), (5), (4), (3), (1)
Pubmed 3 references available
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Quantity 0.1 mL
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Immunogen A synthetic peptide made to a C-terminal portion of the human Aromatase protein (between residues 400-502). Immunogen has 91% homology to horse, 90% homology to cow and rabbit, 83% homology to pig, and 81% homology to sheep, goat, and dog proteins.
Purification affinity purified
Alternative Name Aromatase (CYP19A1 Antibody Abstract)
Background Aromatase is a key enzyme in steroidogenesis and plays an important role in sexualdifferentiation, fertility, and carcinogenesis. Many environmental chemicals mayinfluence aromatase activity and thereby disrupt endocrine function. Alternate Names: anti-ARO1 antibody, anti-CYAR antibody, anti-CYP19 antibody, anti-CYP19A1 antibody,anti-CYPXIX antibody, anti-Cytochrome P450 19A1 antibody, anti-Estrogen synthetaseantibody, anti-P-450AROM antibody.
Gene Symbol: CYP19A1
Gene ID 1588
UniProt P11511
Research Area Cancer, Endocrine system, Enzymes
Application Notes This Aromatase antibody is useful for Immunocytochemistry/Immunofluorescence and Western blot. In Western blot a band is seen at ~55 kDa in human brain, and Immunocytochemistry/Immunofluorescence where membrane staining is observed in SH-SY-5Y cells.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:100, Western Blot 1 µg/mL
Protocol Protocol specific for Aromatase Antibody Western Blot Protocol
1. Perform SDS-PAGE (4-12 %) on samples to be analyzed, loading 20 µg of total protein per lane.
. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
. Rinse membrane with dH2O and then stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
. Rinse the blot in TBS for approximately 5 minutes.
. Block the membrane using 5 % non-fat dry milk + 1 % BSA in TBS for 2 hours at room temperature.
. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.
. Dilute the rabbit anti-Aromatase primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.
. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce's ECL). Viewed band in 1-2 minutes of exposure.Note: Tween-20 can be added to the blocking or antibody diultion buffer at a final concentration of 0.05-0.2 %, provided it does not interfere with antibody-antigen binding.Immunocytochemistry Protocol Immunocytochemistry ProtocolCulture cells to appropriate density in 35 mm culture dishes or 6-well plates.
. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer Tris-citrate/Phosphate, pH 7-8, Sodium Azide
Preservative Sodium azide
Precaution of Use 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.
Handling Advice Do not freeze.
Storage 4 °C
Supplier Images
Western Blotting (WB) image for anti-Cytochrome P450, Family 19, Subfamily A, Polypeptide 1 (CYP19A1) (AA 400-502), (C-Term) antibody (ABIN151143) Detection of aromatasein human fetal temporal lobe lysate using ABIN151143. ECL expos...
Product cited in: Bogan, Murphy, Hennebold: "Dynamic changes in gene expression that occur during the period of spontaneous functional regression in the rhesus macaque corpus luteum." in: Endocrinology, Vol. 150, Issue 3, pp. 1521-9, 2009 (PubMed).

Bogan, Murphy, Stouffer et al.: "Systematic determination of differential gene expression in the primate corpus luteum during the luteal phase of the menstrual cycle." in: Molecular endocrinology (Baltimore, Md.), Vol. 22, Issue 5, pp. 1260-73, 2008 (PubMed).

Background publications Beyer, Green, Barker et al.: "Aromatase-immunoreactivity is localised specifically in neurones in the developing mouse hypothalamus and cortex." in: Brain research, Vol. 638, Issue 1-2, pp. 203-10, 1994 (PubMed).

Catalog No. ABIN151143

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