Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
  • Immunocytochemistry/ Immunofluorescence - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
  • Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
  • Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
  • Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
  • Immunocytochemistry/ Immunofluorescence - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
  • Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
  • Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7739] to Methylmalonyl Coenzyme A mutase
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739]
    See all Methylmalonyl Coenzyme A mutase primary antibodies

  • Description

    Rabbit monoclonal [EPR7739] to Methylmalonyl Coenzyme A mutase

  • Host species

    Rabbit

  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human Methylmalonyl Coenzyme A mutase aa 550-650. The exact sequence is proprietary.
    Database link: P22033

  • Positive control

    • WB: NIH 3T3, K562, 293T, HEK-293, PC-12, C6, mouse kidney, rat kidney, human fetal liver and HeLa cell lysates IHC-P: Human kidney and livertissue ICC/IF: HeLa cells

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133672 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
WB
Mouse
Rat
Human
Application Abreviews Notes
WB
1/1000. Detects a band of approximately 78 kDa (predicted molecular weight: 83 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
ICC/IF
1/100 - 1/250.
Notes
WB
1/1000. Detects a band of approximately 78 kDa (predicted molecular weight: 83 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
ICC/IF
1/100 - 1/250.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      Involved in the degradation of several amino acids, odd-chain fatty acids and cholesterol via propionyl-CoA to the tricarboxylic acid cycle. MCM has different functions in other species.

    • Involvement in disease

      Defects in MUT are the cause of methylmalonic aciduria type mut (MMAM) [MIM:251000]. MMAM is an often fatal disorder of organic acid metabolism. Common clinical features include lethargy, vomiting, failure to thrive, hypotonia, neurological deficit and early death. Two forms of the disease are distinguished by the presence (mut-) or absence (mut0) of residual enzyme activity. Mut0 patients have more severe neurological manifestations of the disease than do MUT- patients. MMAM is unresponsive to vitamin B12 therapy.

    • Sequence similarities

      Belongs to the methylmalonyl-CoA mutase family.
      Contains 1 B12-binding domain.

    • Cellular localization

      Mitochondrion matrix.
    • Information by UniProt

    • Database links

      • Alternative names

        • MCM antibody
        • Methylmalonyl CoA isomerase antibody
        • Methylmalonyl CoA mutase mitochondrial antibody
        • Methylmalonyl Coenzyme A mutase antibody
        • Methylmalonyl-CoA isomerase antibody
        • Methylmalonyl-CoA mutase antibody
        • mitochondrial antibody
        • Mut antibody
        • MUTA_HUMAN antibody
        see all

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling Methylmalonyl Coenzyme A mutase with Purified ab133672 at 1:100 dilution (8.7 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0)Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
      • Immunocytochemistry/ Immunofluorescence - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Immunocytochemistry/ Immunofluorescence - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Methylmalonyl Coenzyme A mutase with Purified ab133672 at 1:100 dilution (9.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor®488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
      • Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        All lanes : Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672) at 0.9 µg/ml (purified)

        Lane 1 : Mouse kidney lysates
        Lane 2 : Rat kidney lysates

        Lysates/proteins at 15 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

        Predicted band size: 83 kDa



        Blocking and diluting buffer: 5% NFDM/TBST.

      • Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        All lanes : Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672) at 0.9 µg/ml (purified)

        Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
        Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
        Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
        Lane 4 : C6 (Rat glial tumor glial cell) whole cell lysates

        Lysates/proteins at 15 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

        Predicted band size: 83 kDa



        Blocking and diluting buffer: 5% NFDM/TBST. 

      • Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)

        Lane 1: Wild-type HAP1 cell lysate (20 µg)
        Lane 2: Methylmalonyl Coenzyme A mutase knockout HAP1 cell lysate (20 µg)
        Lane 3: HeLa cell lysate (20 µg)
        Lane 4: K562 cell lysate (20 µg)

        Lanes 1 - 4: Merged signal (red and green). Green - ab133672 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.

        Unpurified ab133672 was shown to specifically react with Methylmalonyl Coenzyme A mutase when Methylmalonyl Coenzyme A mutase knockout samples were used. Wild-type and Methylmalonyl Coenzyme A mutase knockout samples were subjected to SDS-PAGE. ab133672 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)

        Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling Methylmalonyl Coenzyme A mutase with unpurified ab133672 at 1/50.

      • Immunocytochemistry/ Immunofluorescence - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Immunocytochemistry/ Immunofluorescence - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)

        Immunofluorescent staining of HeLa cells labelling Methylmalonyl Coenzyme A mutase with unpurified ab133672 at 1/100.

      • Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        All lanes : Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672) at 1/1000 dilution (Unpurified)

        Lane 1 : NIH 3T3 cell lysate
        Lane 2 : K562 cell lysate
        Lane 3 : 293T cell lysate
        Lane 4 : HeLa cell lysate
        Lane 5 : Human fetal liver lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

        Predicted band size: 83 kDa

      • Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)
        Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)

      References (1)

      Publishing research using ab133672? Please let us know so that we can cite the reference in this datasheet.

      ab133672 has been referenced in 1 publication.

      • Dalton WB  et al. Hotspot SF3B1 mutations induce metabolic reprogramming and vulnerability to serine deprivation. J Clin Invest 129:4708-4723 (2019). PubMed: 31393856

      Customer reviews and Q&As

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      Immunohistochemistry (Frozen sections) abreview for Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739]

       Excellent
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Frozen sections)
      Sample
      Mouse Tissue sections (Heart (Embryonic))
      Permeabilization
      Yes - Tween-20
      Specification
      Heart (Embryonic)
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
      Fixative
      Paraformaldehyde
      Read More

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      Submitted Aug 29 2016

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