Product Info Summary
SKU: | M00464 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Mouse |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-COMT Antibody Picoband® (monoclonal, 15C10)
SKU/Catalog Number
M00464
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-COMT Antibody Picoband® (monoclonal, 15C10) catalog # M00464. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-COMT Antibody Picoband® (monoclonal, 15C10) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00464)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
15C10
Isotype
Mouse IgG2b
Immunogen
E.coli-derived human COMT recombinant protein (Position: G52-P271). Human COMT shares 81.9% and 81% amino acid (aa) sequence identity with mouse and rat COMT, respectively.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M00464 is reactive to COMT in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
Observed Molecular Weight
29 kDa
Calculated molecular weight
30.037kDa
Background of COMT
Catechol O-methyltransferase, also called COMT, is one of the major mammalian enzymes involved in the metabolic degradation of catecholamines. This gene is mapped to 22q11.21. Catechol-O-methyltransferase catalyzes the transfer of a methyl group from S-adenosylmethionine to catecholamines, including the neurotransmitters dopamine, epinephrine, and norepinephrine. This O-methylation results in one of the major degradative pathways of the catecholamine transmitters. In addition to its role in the metabolism of endogenous substances, COMT is important in the metabolism of catechol drugs used in the treatment of hypertension, asthma, and Parkinson disease. COMT is found in two forms in tissues, a soluble form (S-COMT) and a membrane-bound form (MB-COMT). The differences between S-COMT and MB-COMT reside within the N-termini.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M00464 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human,Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human HepG2 whole cell, human K562 whole cell, human THP-1 whole cell, human HEK293 whole cell, human A549 whole cell, human Caco-2 whole cell, rat RH35 whole cell, mouse Neuro-2a whole cell,
IHC: human intestinal cancer tissue, human mammary cancer tissue, human mammary cancer tissue, human placenta tissue, human lung cancer tissue, mouse testis tissue, rat testis tissue
ICC/IF: A431 cell, MCF7 cell
FCM: K562 cell, U87 cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of COMT using anti-COMT antibody (M00464).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HepG2 whole cell lysates
Lane 2: human K562 whole cell lysates
Lane 3: human THP-1 whole cell lysates
Lane 4: human HEK293 whole cell lysates
Lane 5: human A549 whole cell lysates
Lane 6: human Caco-2 whole cell lysates
Lane 7: rat RH35 whole cell lysates
Lane 8: mouse Neuro-2a whole cell lysates
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-COMT antigen affinity purified monoclonal antibody (Catalog # M00464) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for COMT at approximately 29KD. The expected band size for COMT is at 29KD.
Click image to see more details
Figure 2. IHC analysis of COMT using anti-COMT antibody (M00464).
COMT was detected in paraffin-embedded section of human intestinal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COMT Antibody (M00464) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of COMT using anti-COMT antibody (M00464).
COMT was detected in paraffin-embedded section of human mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COMT Antibody (M00464) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of COMT using anti-COMT antibody (M00464).
COMT was detected in paraffin-embedded section of human mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COMT Antibody (M00464) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of COMT using anti-COMT antibody (M00464).
COMT was detected in paraffin-embedded section of human placenta tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COMT Antibody (M00464) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of COMT using anti-COMT antibody (M00464).
COMT was detected in paraffin-embedded section of human lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COMT Antibody (M00464) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 7. IHC analysis of COMT using anti-COMT antibody (M00464).
COMT was detected in paraffin-embedded section of mouse testis tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COMT Antibody (M00464) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 8. IHC analysis of COMT using anti-COMT antibody (M00464).
COMT was detected in paraffin-embedded section of rat testis tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COMT Antibody (M00464) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 9. IF analysis of COMT using anti-COMT antibody (M00464).
COMT was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-COMT Antibody (M00464) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 10. Flow Cytometry analysis of K562 cells using anti-COMT antibody (M00464).
Overlay histogram showing K562 cells stained with M00464 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-COMT Antibody (M00464,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Click image to see more details
Figure 11. Flow Cytometry analysis of U87 cells using anti-COMT antibody (M00464).
Overlay histogram showing U87 cells stained with M00464 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-COMT Antibody (M00464,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Click image to see more details
Figure 12. IF analysis of COMT using anti-COMT antibody (M00464).
COMT was detected in immunocytochemical section of MCF7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-COMT Antibody (M00464) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Protein Target Info & Infographic
Gene/Protein Information For COMT (Source: Uniprot.org, NCBI)
Gene Name
COMT
Full Name
Catechol O-methyltransferase
Weight
30.037kDa
Superfamily
class I-like SAM-binding methyltransferase superfamily
Alternative Names
Catechol O-methyltransferase; COMT COMT HEL-S-98n catechol-O-methyltransferase catechol O-methyltransferase|epididymis secretory sperm binding protein Li 98n|testicular tissue protein Li 42
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on COMT, check out the COMT Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for COMT: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-COMT Antibody Picoband® (monoclonal, 15C10) (M00464)
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