Overview

  • Product name
    Anti-ACADM/MCAD antibody [EPR3708]
    See all ACADM/MCAD primary antibodies
  • Description
    Rabbit monoclonal [EPR3708] to ACADM/MCAD
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ACADM/MCAD aa 200-300. The exact sequence is proprietary.

  • Positive control
    • WB: Human heart or fetal liver tissue lysates; HeLa, HepG2, or K562 cell lysates. IHC-P: Human liver tissue. ICC/IF: HeLa cells.
  • General notes

    Previously labelled as ACADM. 

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab92461 in the following tested applications.

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

Application Abreviews Notes
WB
IP
IHC-P
ICC/IF
Flow Cyt
  • Application notes
    ICC/IF: 1/100 - 1/250.
    IHC-P: 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. The use of an HRP/AP polymerized secondary antibody is recommended.
    IP: 1/10 - 1/100.
    WB: 1/10000 - 1/50000. Predicted molecular weight: 47 kDa.

    Is unsuitable for Flow Cyt.

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      This enzyme is specific for acyl chain lengths of 4 to 16.
    • Pathway
      Lipid metabolism; mitochondrial fatty acid beta-oxidation.
    • Involvement in disease
      Defects in ACADM are the cause of acyl-CoA dehydrogenase medium-chain deficiency (ACADMD) [MIM:201450]. It is an autosomal recessive disease which causes fasting hypoglycemia, hepatic dysfunction, and encephalopathy, often resulting in death in infancy.
    • Sequence similarities
      Belongs to the acyl-CoA dehydrogenase family.
    • Cellular localization
      Mitochondrion matrix.
    • Information by UniProt
    • Database links
    • Alternative names
      • ACAD 1 antibody
      • ACAD1 antibody
      • Acadm antibody
      • ACADM_HUMAN antibody
      • Acyl coenzyme A dehydrogenase antibody
      • Acyl coenzyme A dehydrogenase C 4 to C 12 straight chain antibody
      • FLJ18227 antibody
      • FLJ93013 antibody
      • FLJ99884 antibody
      • MCAD antibody
      • MCADH antibody
      • Medium chain acyl CoA dehydrogenase antibody
      • Medium chain fatty acyl CoA dehydrogenase antibody
      • Medium chain specific acyl CoA dehydrogenase antibody
      • Medium chain specific acyl CoA dehydrogenase mitochondrial antibody
      • Medium-chain specific acyl-CoA dehydrogenase antibody
      • mitochondrial antibody
      see all

    Images

    • All lanes : Anti-ACADM/MCAD antibody [EPR3708] (ab92461) at 1/10000 dilution

      Lane 1 : Human heart lysate
      Lane 2 : fetal liver lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : HepG2 cell lysate
      Lane 5 : K562 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP conjugated Goat anti-Rabbit Ig at 1/2000 dilution

      Predicted band size: 47 kDa

    • ab92461, at 1/100 dilution, staining ACADM/MCAD in formalin-fixed, paraffin-embedded Human liver tissue by immunohistochemistry.

    • Immunofluorescent staining of ACADM/MCAD in HeLa cells using ab92461 at 1/100 dilution.

    • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ACADM/MCAD with unpurified ab92461 at 1/50 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

    References

    This product has been referenced in:
    • Zheng F & Cai Y Concurrent exercise improves insulin resistance and nonalcoholic fatty liver disease by upregulating PPAR-? and genes involved in the beta-oxidation of fatty acids in ApoE-KO mice fed a high-fat diet. Lipids Health Dis 18:6 (2019). Read more (PubMed: 30611282) »
    • Peng J  et al. The detrimental effects of glucocorticoids exposure during pregnancy on offspring's cardiac functions mediated by hypermethylation of bone morphogenetic protein-4. Cell Death Dis 9:834 (2018). Read more (PubMed: 30082698) »
    See all 7 Publications for this product

    Customer reviews and Q&As

    1-8 of 8 Abreviews or Q&A

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (human hepatocytes)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    15 µg
    Specification
    human hepatocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

    Abcam user community

    Verified customer

    Submitted Dec 28 2017

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HEK293)
    Specification
    HEK293
    Blocking step
    Serum as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Apr 03 2017

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing (10% SDS PAGE)
    Loading amount
    30 µg
    Specification
    Brain
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted May 27 2014

    Answer

    I appreciate the time that has been spent on these experiments and would be pleased to arrange, in compensation, a replacement of ab92461 with another anti-ACADM such as ab110296. Alternatively I can also arrange a credit note. I look forward to hearing from you with details of how you would like to proceed. Could you also please send me one of the following : the name of the distributor and the date of purchase? www.abcam.com/ab110296

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    Question
    Answer

    Thank you very much for your email. My colleague is in contact with the laboratory in order to try to find out what the problem might is. Indeed we have to investigate this issue further as the quality of our products is most important. We have had no reason so far to suspect any problems with this antibody. Can you confirm also that using no primary antibody - or an alternative rabbit antibody - the secondary antibody alone does not give any signal? Thank you very much for your cooperation. I would like to apologize for the long delays to resolve this issue. In order to allow you to continue with your research meanwhile, I can offer you an alternative antibody against ACADM. I would like to suggest the ab110296, Anti-ACADM antibody [ clone 3B7BH7], a mouse monoclonal antibody tested in WB and on mouse, rat, cow and human. The specificity of this antibody has also been validated with mass spectrometry. https://www.abcam.com/index.html?datasheet=110296 Please let me know whether you would like to accept the alternative antibody. I am looking forward to hear back from you.

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    Answer

    Thank you for sending the image. We can’t make assumptions for the specificity of an antibody because of its same size band specificity of as of beta actin. Many protein have similar sizes, so experimental evidence is always good to prove the cross reactivity of any ab. I have following recommendations; - Use beta actin negative control lysate e.g. skeletal muscle myocyte and heart muscle myocytes are negative for beta actin so the lysates of these will be a good positive control. ab29330; https://www.abcam.com/Skeletal-Muscle-Human-Tissue-Lysate-adult-normal-tissue-ab29330.html http://www.proteinatlas.org/ENSG00000075624/normal - There is no sequence similarity between ACADM and beta actin so the antibody can’t cross reacts with beta actin. I would suggest using purified beta actin protein as a positive control. http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111102-154040-0003-65312871-pg - ACADM is generally present in mitochondrial matrix. Prepare the mitochondrial lysates by getting rid of beta actin to see if the antibody was able to detect the ACADM or not. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. PS: Let me know if you are interested in using lysates ab29330; I can offer you 50% discount.

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    Question
    Answer

    Thank you for contacting us. The antibody is specific against ACADM protein only; ab92461 should not recognize the beta actin protein. Could you please explain why you think the antibody is detecting the beta actin protein? In regards to the expected band size of ACADM which is 47 kDa, that may run at different molecular weight due to various post translational modifications however we can’t say the ab92461is detecting beta actin instead of ACADM. I would suggest using beta actin negative lysates as a control lysates for experimental proof e.g. heart muscle myocytes are negative for beta actin. Looking froward to hearing from you. Have a good day!

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    Answer

    Thank you for your enquiry regarding ab92461 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody. I appreciate the time you have spent on these experiments in the laboratory and it is disappointing the results have not been successful. To identify the source of the problem I would like to ask few questions; - Was the lysates denatured in loading buffer? We normally recommend our customers using 1X SDS loading buffer with boiling at 100C for 5 minutes. - Could you provide an image? Regarding the band below 42 kDa; is this the only band observed? - Could you specify at what temperature the tissues were treated with RIPA buffer? Also give a breif description e.g. how the lysates were prepared? - Does the band resolved properly in gel? - Finally could you provide the Abcam order number? The order number give in your email does not match any order record for this product. I will look forward to receiving your reply soon.

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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