Name: Anti-ACADM/MCAD antibody [EPR3708]
Brand: RabMAb
SKU: ab92461
Rating: 5 (7 reviews)

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Western blot - 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

Alexa Fluor® 488 Carrier Free HRP

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.

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

Alternative Versions
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)

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.

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

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) 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 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)

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)

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)

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) 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) 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)

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)

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)

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

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

Click here to view the general protocols

Datasheet download

Download

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

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

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

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

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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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Conjugation kits

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Pathway

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (16)

Customer reviews and Q&As

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

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