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Anti-Methylmalonyl Coenzyme A mutase Antibody Picoband™ (monoclonal, 2D6)

Figure 1. Western blot analysis of Methylmalonyl Coenzyme A using anti-Methylmalonyl Coenzyme A antibody (M01065).
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 Hela whole cell lysates;
Lane 2: human K562 whole cell lysates;
Lane 3: human HEK293 whole cell lysates;
Lane 4: human PC-3 whole cell lysates;
Lane 5: human Caco-2 whole cell lysates;
Lane 6: human Raji whole cell lysates;
Lane 7: rat PC-12 whole cell lysates;
Lane 8: mouse NIH/3T3 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-Methylmalonyl Coenzyme A antigen affinity purified monoclonal antibody (Catalog # M01065) 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 Methylmalonyl Coenzyme A at approximately 83KD. The expected band size for Methylmalonyl Coenzyme A is at 83KD.
Figure 2. IHC analysis of Methylmalonyl Coenzyme A using anti-Methylmalonyl Coenzyme A antibody (M01065).
Methylmalonyl Coenzyme A was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Methylmalonyl Coenzyme A Antibody (M01065) 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.
Figure 3. IHC analysis of Methylmalonyl Coenzyme A using anti-Methylmalonyl Coenzyme A antibody (M01065).
Methylmalonyl Coenzyme A was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Methylmalonyl Coenzyme A Antibody (M01065) 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.
Figure 4. IHC analysis of Methylmalonyl Coenzyme A using anti-Methylmalonyl Coenzyme A antibody (M01065).
Methylmalonyl Coenzyme A was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Methylmalonyl Coenzyme A Antibody (M01065) 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.
Figure 5. IHC analysis of Methylmalonyl Coenzyme A using anti-Methylmalonyl Coenzyme A antibody (M01065).
Methylmalonyl Coenzyme A was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Methylmalonyl Coenzyme A Antibody (M01065) 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.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC
Primary Accession	P22033
Host	Mouse
Isotype	Mouse IgG2b
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-Methylmalonyl Coenzyme A mutase Antibody Picoband™ (monoclonal, 2D6) . Tested in IHC, WB applications. This antibody reacts with Human, Mouse, Rat.
Reconstitution	Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Additional Information
Gene ID	4594
Other Names	Methylmalonyl-CoA mutase, mitochondrial, MCM, 5.4.99.2, Methylmalonyl-CoA isomerase, MMUT (HGNC:7526)
Calculated MW	83134 Da
Application Details	Western blot, 0.1-0.5 µg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1 µg/ml, Human, Rat
Subcellular Localization	Mitochondrion.
Source	Eukaryota
Contents	Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na₂HPO₄, 0.05mg NaN₃.
Clone Names	Clone: 2D6
Immunogen	A synthetic peptide corresponding to a sequence at the N-terminus of human MUT, different from the related mouse sequence by one amino acid.
Purification	Immunogen affinity purified.
Cross Reactivity	No cross-reactivity with other proteins.
Storage	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.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	MMUT (HGNC:7526)
Function	Catalyzes the reversible isomerization of methylmalonyl-CoA (MMCoA) (generated from branched-chain amino acid metabolism and degradation of dietary odd chain fatty acids and cholesterol) to succinyl-CoA (3-carboxypropionyl-CoA), a key intermediate of the tricarboxylic acid cycle.
Cellular Location	Mitochondrion matrix. Mitochondrion. Cytoplasm

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

Methylmalonyl-CoA mutase (MUT) is a mitochondrial enzyme that catalyzes the isomerization of methylmalonyl-CoA to succinyl-CoA. This gene is mapped to 6p12.3. MUT requires a vitamin B12-derived prosthetic group, adenosylcobalamin (commonly referred to as AdoCbl), to function. And the product of this enzyme, succinyl-CoA, is a key molecule of the TCA cycle.

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¥ 3,367.00

Cat# ABO14958

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Anti-Methylmalonyl Coenzyme A mutase Antibody Picoband™ (monoclonal, 2D6)

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