Recombinant
RabMAb

Anti-FACL4 antibody [EPR8640] (ab155282)

Overview

  • Product name
    Anti-FACL4 antibody [EPR8640]
    See all FACL4 primary antibodies
  • Description
    Rabbit monoclonal [EPR8640] to FACL4
  • Tested applications
    Suitable for: IHC-P, IP, WB, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human FACL4.

  • Positive control
    • WB: HepG2, HeLa and 293T cell lysates and human fetal kidney tissue, mouse brain tissue, rat brain, kidney and spleen tissue lysates. IHC-P: human hepatocellular carcinoma tissue. ICC/IF: HeLa cells. Flow Cyt: Permeabilized 293T cells. IP: HepG2 cell lysate.
  • General notes

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab155282 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
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

IP 1/10 - 1/100.
WB 1/10000 - 1/50000. Predicted molecular weight: 79 kDa.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100 - 1/250.

Target

  • Function
    Activation of long-chain fatty acids for both synthesis of cellular lipids, and degradation via beta-oxidation. Preferentially uses arachidonate and eicosapentaenoate as substrates.
  • Involvement in disease
    Defects in ACSL4 are the cause of mental retardation X-linked type 63 (MRX63) [MIM:300387]. Mental retardation is a mental disorder characterized by significantly sub-average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. Non-syndromic mental retardation patients do not manifest other clinical signs.
    Defects in ACSL4 are involved in Alport syndrome with mental retardation midface hypoplasia and elliptocytosis (ATS-MR) [MIM:300194]. A X-linked contiguous gene deletion syndrome characterized by glomerulonephritis, deafness, mental retardation, midface hypoplasia and elliptocytosis.
  • Sequence similarities
    Belongs to the ATP-dependent AMP-binding enzyme family.
  • Cellular localization
    Mitochondrion outer membrane. Peroxisome membrane. Microsome membrane. Endoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ACS 4 antibody
    • ACS4 antibody
    • ACSL 4 antibody
    • Acsl4 antibody
    • ACSL4_HUMAN antibody
    • acyl CoA synthetase 4 antibody
    • Acyl CoA synthetase long chain family member 4 antibody
    • FACL 4 antibody
    • FACL4 antibody
    • Fatty acid Coenzyme A ligase antibody
    • fatty acid Coenzyme A ligase long-chain 4 antibody
    • LACS 4 antibody
    • LACS4 antibody
    • Lignoceroyl CoA synthase antibody
    • Long chain 4 antibody
    • long chain acyl CoA synthetase 4 antibody
    • long chain fatty acid CoA ligase 4 antibody
    • long chain fatty acid Coenzyme A ligase 4 antibody
    • Long-chain acyl-CoA synthetase 4 antibody
    • Long-chain-fatty-acid--CoA ligase 4 antibody
    • MRX63 antibody
    • MRX68 antibody
    see all

Images

  • All lanes : Anti-FACL4 antibody [EPR8640] (ab155282) at 1/1000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 79 kDa
    Observed band size : 79 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Anti-FACL4 antibody [EPR8640] (ab155282) at 1/50000 dilution (purified) + HepG2 whole cell lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 79 kDa
    Observed band size : 79 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-FACL4 antibody [EPR8640] (ab155282) at 1/10000 dilution (purified)

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HEK293 whole cell lysate
    Lane 3 : Human fetal kidney tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugate goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 79 kDa
    Observed band size : 79 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue labelling FACL4 with purified ab155282 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling FACL4 with purified ab155282 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/200) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Flow Cytometry analysis of 293T cells labelling FACL4 with purified ab155282 at 1/100 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • ab155282 (purified) at 1/30 immunoprecipitating FACL4 in HepG2 whole cell lysate.

    Lane 1 (input): HepG2 whole cell lysate (10µg)

    Lane 2 (+): ab155282 + HepG2 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab155282 in HepG2 whole cell lysate.

    For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-FACL4 antibody [EPR8640] (ab155282) at 1/10000 dilution (unpurified)

    Lane 1 : HepG2 lysate
    Lane 2 : HeLa lysate
    Lane 3 : 293T lysate
    Lane 4 : Fetal kidney lysate, human

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 79 kDa
  • All lanes : Anti-FACL4 antibody [EPR8640] (ab155282) at 1/10000 dilution (unpurified)

    Lane 1 : Rat brain tissue lysate
    Lane 2 : Rat heart tissue lysate
    Lane 3 : Rat kidney tissue lysate
    Lane 4 : Rat spleen tissue lysate


    Predicted band size : 79 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • Flow cytometric analysis of permeabilized 293T cells labelling FACL4 with unpurified ab155282 at a dilution of 1/10 (red) compared to a rabbit IgG (negative) (green).

References

This product has been referenced in:

See 1 Publication for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
human hepatocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted May 31 2017

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