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Purified Mouse Anti-COMT
Purified Mouse Anti-COMT

Western blot analysis of COMT on a rat pituitary lysate (left). Lane 1: 1:10,000, lane 2: 1:20,000, lane 3: 1:40,000 dilution of the anti- COMT antibody.

Immunofluorescent staining of C6 cells (right). Cells were seeded in a 384 well collagen coated Microplates (Material # 353962) at ~ 6,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure; Bioimaging protocol link) and the anti-COMT antibody. The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen)(pseudo colored green). Cell nuclei were counter stained with Hoechst 33342 (pseudo colored blue). The image was taken on a BD Pathway™  855 or 435 Bioimager System using a 20x objective and merged using the BD AttoVison ™ software.This antibody stained C6 cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link), and only reacts with rat cells.

Western blot analysis of COMT on a rat pituitary lysate (left). Lane 1: 1:10,000, lane 2: 1:20,000, lane 3: 1:40,000 dilution of the anti- COMT antibody.

Immunofluorescent staining of C6 cells (right). Cells were seeded in a 384 well collagen coated Microplates (Material # 353962) at ~ 6,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure; Bioimaging protocol link) and the anti-COMT antibody. The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen)(pseudo colored green). Cell nuclei were counter stained with Hoechst 33342 (pseudo colored blue). The image was taken on a BD Pathway™  855 or 435 Bioimager System using a 20x objective and merged using the BD AttoVison ™ software.This antibody stained C6 cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link), and only reacts with rat cells.

Product Details
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BD Transduction Laboratories™
Catechol-O-Methyltransferase
Rat (QC Testing), Mouse (Tested in Development)
Mouse IgG1
Mouse COMT aa. 26-141
Western blot (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
24/28 kDa
250 µg/ml
AB_399391
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at -20°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
  6. Triton is a trademark of the Dow Chemical Company.
  7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
611970 Rev. 2
Antibody Details
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4/COMT

Catechol-O-methyltransferase (COMT) and monoamine oxidase (MAO) are the major mammalian enzymes involved in the degradation of the catecholamine neurotransmitters, dopamine, norepinephrine, and epinephrine. COMT is a Mg2+-dependent enzyme that catalyzes the transfer of methyl groups from S-adenosyl methionine to a hydroxyl group of a catecholic substrate. Two forms of COMT are found in rat brain, a 24 kDa soluble COMT (S-COMT) and a 28 kDa membrane-bound COMT (MB-COMT). COMT is widely expressed in brain, but its importance in catecholamine neurotransmitter degradation relative to MAO varies in different brain regions. In addition, COMT may function primarily in extraneuronal areas, such as in glial cells and postsynaptic neurons. COMT-deficient mice have sex- and region-specific alterations in dopamine levels in the brain, and display impaired emotional reactivity and aggressive behavior. Thus, COMT-mediated degradation of catecholamines in the brain may have important roles in maintaining normal catecholamine levels, as well as normal social behavior.

611970 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
611970 Rev.2
Citations & References
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Development References (3)

  1. Gogos JA, Morgan M, Luine V, et al. Catechol-O-methyltransferase-deficient mice exhibit sexually dimorphic changes in catecholamine levels and behavior. Proc Natl Acad Sci U S A. 1998; 95(17):9991-9996. (Biology). View Reference
  2. Tilgmann C, Melen K, Lundstrom K, et al. Expression of recombinant soluble and membrane-bound catechol O-methyltransferase in eukaryotic cells and identification of the respective enzymes in rat brain. Eur J Biochem. 1992; 207(2):813-821. (Biology). View Reference
  3. Werner P, Di Rocco A, Prikhojan A, et al. COMT-dependent protection of dopaminergic neurons by methionine, dimethionine and S-adenosylmethionine (SAM) against L-dopa toxicity in vitro. Brain Res. 2001; 893(1-2):278-281. (Biology). View Reference
611970 Rev. 2

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