COMT Polyclonal antibody

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  WB analysis using 14754-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 14754-1-AP (COMT antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  IP experiment of K-562 using 14754-1-AP

  IP result of anti-COMT (IP:14754-1-AP, 4ug; Detection:14754-1-AP 1:500) with K-562 cells lysate 1200ug.

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  IHC staining of human liver using 14754-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver using 14754-1-AP (COMT antibody) at dilution of 1:100 (under 40x lens).

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  IHC staining of human placenta using 14754-1-AP

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 14754-1-AP (COMT antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human placenta using 14754-1-AP

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 14754-1-AP (COMT antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse lung using 14754-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 14754-1-AP (COMT antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse lung using 14754-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 14754-1-AP (COMT antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of mouse cerebellum using 14754-1-AP

  Immunofluorescent analysis of (4% PFA) fixed mouse cerebellum tissue using COMT antibody (14754-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of mouse cerebellum using 14754-1-AP

  Immunofluorescent analysis of (4% PFA) fixed mouse cerebellum tissue using COMT antibody (14754-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of HepG2 using 14754-1-AP

  Immunofluorescent analysis of HepG2 cells, using COMT antibody 14754-1-AP at 1:50 dilution and Rhodamine-labeled goat anti-rabbit IgG (red). Blue pseudocolor = DAPI (fluorescent DNA dye).

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  FC experiment of PC-3 using 14754-1-AP

  1x10^6 PC-3 cells were intracellularly stained with 0.8 ug COMT Polyclonal antibody (14754-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.8 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

COMT Polyclonal Antibody for WB, IHC, IF/ICC, IF-P, FC (Intra), IP, ELISA

Cat No. 14754-1-AP

Publications(13)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (1)

Applications

WB, IHC, IF/ICC, IF-P, FC (Intra), IP, ELISA

Synonyms

catechol O methyltransferase, Catechol O-methyltransferase, EC:2.1.1.6

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Tested Applications

• Positive WB detected in: HEK-293 cells, K-562 cells, human placenta, mouse liver, rat liver
• Positive IP detected in: K-562 cells
• Positive IHC detected in: human liver tissue, human placenta tissue, mouse lung tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: mouse cerebellum tissue
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: PC-3 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:4000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:100-1:800
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.80 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 11 publications below
• IHC: See 2 publications below
• IF: See 3 publications below

Product Information

14754-1-AP targets COMT in WB, IHC, IF/ICC, IF-P, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, monkey
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: COMT fusion protein Ag6502
• Full Name: catechol-O-methyltransferase
• Calculated Molecular Weight: 30 kDa
• Observed Molecular Weight: 28-30 kDa
• GenBank Accession Number: BC000419
• Gene Symbol: COMT
• Gene ID (NCBI): 1312
• RRID: AB_2083706
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P21964
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Catechol O-methyltransferase (COMT) is an important enzyme that catalyzes O-methylation and inactivation of metabolically active catechol-containing bioactive molecules and drugs. The mammalian COMT enzyme exists in soluble form (24-26 kDa) and membrane-bound form (28-30 kDa), which are encoded by a single gene utilizing different promoters.

Publications

Species	Application	Title
mouse	WB	Aging (Albany NY); A five-m6A regulatory gene signature is a prognostic biomarker in lung adenocarcinoma patients.; Authors - Xiao Wu
mouse	WB	Aging (Albany NY); Maternal high sugar and fat diet benefits offspring brain function via targeting on the gut-brain axis.; Authors - Dongdong Wang
mouse	WB	Transl Psychiatry; A diet high in sugar and fat influences neurotransmitter metabolism and then affects brain function by altering the gut microbiota.; Authors - Yinrui Guo
mouse	WB	FASEB J; Entacapone alleviates acute kidney injury by inhibiting ferroptosis.; Authors - Jiahong Yang
human	WB,IHC	Endocrinology; Norepinephrine, active norepinephrine transporter, and norepinephrine-metabolism are involved in the generation of reactive oxygen species in human ovarian granulosa cells.; Authors - Saller S S
human	IHC	J Proteomics; Proteomic analysis of podocyte exosome-enriched fraction from normal human urine.; Authors - Prunotto Marco M