Recombinant
RabMAb

Anti-Galactosidase alpha antibody [EP5828(2)] (ab168341)

Overview

  • Product name
    Anti-Galactosidase alpha antibody [EP5828(2)]
    See all Galactosidase alpha primary antibodies
  • Description
    Rabbit monoclonal [EP5828(2)] to Galactosidase alpha
  • Tested applications
    Suitable for: Flow Cyt, IP, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Galactosidase alpha aa 100-200.
    Database link: P06280

  • Positive control
    • WB: MCF-7, 293T, A431 and HeLa cell lysates. IHC-P: Human urinary bladder carcinoma, kidney and uterus tissue. ICC/IF: HeLa cells IP: MCF-7 cell lysates. Flow Cyt: HeLa cells.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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

  • 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
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EP5828(2)
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab168341 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
Flow Cyt 1/90.
IP 1/10 - 1/100.
WB 1/1000 - 1/10000. Predicted molecular weight: 49 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

ICC/IF 1/50 - 1/500.

 

Target

  • Involvement in disease
    Defects in GLA are the cause of Fabry disease (FD) [MIM:301500]. FD is a rare X-linked sphingolipidosis disease where glycolipid accumulates in many tissues. The disease consists of an inborn error of glycosphingolipid catabolism. FD patients show systemic accumulation of globotriaoslyceramide (Gb3) and related glycosphingolipids in the plasma and cellular lysosomes throughout the body. Clinical recognition in males results from characteristic skin lesions (angiokeratomas) over the lower trunk. Patients may show ocular deposits, febrile episodes, and burning pain in the extremities. Death results from renal failure, cardiac or cerebral complications of hypertension or other vascular disease. Heterozygous females may exhibit the disorder in an attenuated form, they are more likely to show corneal opacities.
  • Sequence similarities
    Belongs to the glycosyl hydrolase 27 family.
  • Cellular localization
    Lysosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • AGAL_HUMAN antibody
    • Agalsidase alfa antibody
    • Alpha D galactosidase A antibody
    • Alpha D galactoside galactohydrolase 1 antibody
    • Alpha D galactoside galactohydrolase antibody
    • Alpha gal A antibody
    • Alpha galactosidase A antibody
    • Alpha-D-galactosidase A antibody
    • Alpha-D-galactoside galactohydrolase antibody
    • Alpha-galactosidase A antibody
    • GALA antibody
    • Galactosidase, alpha antibody
    • GLA antibody
    • GLA protein antibody
    • Melibiase antibody
    see all

Images

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Galactosidase with purified ab168341 at 1/500. Cells were fixed with 100% methanol 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 (1/500) 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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human uterus tissue labelling Galactosidase alpha with unpurified ab168341 at a dilution of 1/50.

  • Anti-Galactosidase alpha antibody [EP5828(2)] (ab168341) at 20 µg (purified) + MCF-7 whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size : 49 kDa
    Observed band size : 46 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Galactosidase alpha with purified ab168341 at 1/50. 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.

  • Flow Cytometry analysis of HeLa cells labelling Galactosidase alpha with purified ab168341 at a dilution of 1/90 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Flour® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • ab168341 (purified) at 1/60 immunoprecipitating Galactosidase alpha in MCF-7 whole cell lysate.

    Lane 1 (input): MCF-7 whole cell lysate (10µg)

    Lane 2 (+): ab168341 + MCF-7 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab168341 in MCF-7 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.

  • Anti-Galactosidase alpha antibody [EP5828(2)] (ab168341) at 1/5000 dilution (purified) + HEK293 whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size : 49 kDa
    Observed band size : 46 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human urinary bladder carcinoma tissue labelling Galactosidase alpha with unpurified ab168341 at a dilution of 1/50.

  • All lanes : Anti-Galactosidase alpha antibody [EP5828(2)] (ab168341) at 1/1000 dilution (unpurified)

    Lane 1 : MCF-7 cell lysates
    Lane 2 : 293T cell lysates
    Lane 3 : A431 cell lysates
    Lane 4 : HeLa cell lysates

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 49 kDa

References

ab168341 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab168341.
Please use the links above to contact us or submit feedback about this product.

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

Sign up