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    acadmmcad-antibody-epr3708-ab92461.pdf

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Signal Transduction Metabolism Mitochondrial
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RecombinantRabMAb

Recombinant Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

  • Datasheet
Reviews (7)Q&A (5)References (16)

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Western blot - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Immunocytochemistry/ Immunofluorescence - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Flow Cytometry (Intracellular) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Western blot - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Immunoprecipitation - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Western blot - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Immunocytochemistry/ Immunofluorescence - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Flow Cytometry (Intracellular) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
  • Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3708] to ACADM/MCAD
  • Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)
  • Reacts with: Mouse, Rat, Human

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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 Cyt (Intra)more 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, mouse liver, and rat stomach tissues; ICC/IF: HeLa cells; IP: Mouse heart lysate; Flow Cyt (intra): 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3708
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Cardiovascular
    • Lipids / Lipoproteins
    • Fatty Acids
    • Metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of lipids and lipoproteins
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipid metabolism
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Fatty acid oxidation

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-ACADM/MCAD antibody [EPR3708] (ab203262)
    • HRP Anti-ACADM/MCAD antibody [EPR3708] (ab203465)
    • Anti-ACADM/MCAD antibody [EPR3708] - BSA and Azide free (ab239914)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human ACADM/MCAD protein (ab99329)

Applications

The Abpromise guarantee

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 (2)
1/10000 - 1/50000. Predicted molecular weight: 47 kDa.
IP
1/10 - 1/100.
IHC-P (1)
1/600. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

ICC/IF (2)
1/50 - 1/250.
Flow Cyt (Intra)
1/60.
Notes
WB
1/10000 - 1/50000. Predicted molecular weight: 47 kDa.
IP
1/10 - 1/100.
IHC-P
1/600. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

ICC/IF
1/50 - 1/250.
Flow Cyt (Intra)
1/60.

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.
  • Target information above from: UniProt accession P11310 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 34 Human
    • Entrez Gene: 11364 Mouse
    • Entrez Gene: 24158 Rat
    • Omim: 607008 Human
    • SwissProt: P11310 Human
    • SwissProt: P45952 Mouse
    • SwissProt: P08503 Rat
    • Unigene: 445040 Human
    • Unigene: 10530 Mouse
    • Unigene: 6302 Rat
    see all
  • 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

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

    Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
    Lane 2 : Human heart lysates
    Lane 3 : Mouse heart lysates
    Lane 4 : Rat heart lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 47 kDa
    Observed band size: 43 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue sections labeling ACADM/MCAD with purified ab92461 at 1/600 dilution (1.02 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Immunocytochemistry/ Immunofluorescence - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ACADM/MCAD with purified ab92461 at 1:50 dilution (10 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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Flow Cytometry (Intracellular) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Flow Cytometry (Intracellular) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

    Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ACADM/MCAD with purified ab92461 at 1/60 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

     

  • Western blot - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Western blot - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Anti-ACADM/MCAD antibody [EPR3708] (ab92461) at 1/10000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

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

    Predicted band size: 47 kDa
    Observed band size: 43 kDa why is the actual band size different from the predicted?

  • Immunoprecipitation - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Immunoprecipitation - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    ab92461 (purified) at 1/30 dilution (2 µg) immunoprecipitating ACADM/MCAD in Mouse heart lysate.
    Lane 1 (input): Mouse heart lysate 10 µg
    Lane 2 (+): ab92461 & Mouse heart lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92461 in Mouse heart lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling ACADM/MCAD with purified ab92461 at 1/600 dilution (1.02 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling ACADM/MCAD with purified ab92461 at 1/600 dilution (1.02 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

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

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

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

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Immunocytochemistry/ Immunofluorescence - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

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

  • Flow Cytometry (Intracellular) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Flow Cytometry (Intracellular) - Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

    Intracellular 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. 

     

     

  • Anti-ACADM/MCAD antibody [EPR3708] (ab92461)
    Anti-ACADM/MCAD antibody [EPR3708] (ab92461)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (16)

Publishing research using ab92461? Please let us know so that we can cite the reference in this datasheet.

ab92461 has been referenced in 16 publications.

  • Raimo S  et al. Mitochondrial morphology, bioenergetics and proteomic responses in fatty acid oxidation disorders. Redox Biol 41:101923 (2021). PubMed: 33725513
  • Tang J  et al. Severe pantothenic acid deficiency induces alterations in the intestinal mucosal proteome of starter Pekin ducks. BMC Genomics 22:491 (2021). PubMed: 34193047
  • Hu X  et al. Toll-like receptor 4 is a master regulator for colorectal cancer growth under high-fat diet by programming cancer metabolism. Cell Death Dis 12:791 (2021). PubMed: 34385421
  • Zhang B  et al. Effects of riboflavin deficiency on the lipid metabolism of duck breeders and duck embryos. Poult Sci 100:101342 (2021). PubMed: 34438327
  • Ma C  et al. piRNA-63076 contributes to pulmonary arterial smooth muscle cell proliferation through acyl-CoA dehydrogenase. J Cell Mol Med 24:5260-5273 (2020). PubMed: 32227582
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-ACADM/MCAD antibody [EPR3708]

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 28 2017

Western blot abreview for Anti-ACADM/MCAD antibody [EPR3708]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 27 2014

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