Overview

  • Product name

  • Description

    Rabbit polyclonal to GAA
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Full length protein, corresponding to amino acids 1-952 of Human GAA.

  • Positive control

    • Mouse intestine tissue lysate. GAA transfected 293T cell lysate. IHC-P: Human placenta FFPE tissue sections

Properties

Applications

Our Abpromise guarantee covers the use of ab102815 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 1/500 - 1/1000. Predicted molecular weight: 105 kDa.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    Essential for the degradation of glygogen to glucose in lysosomes.
  • Involvement in disease

    Defects in GAA are the cause of glycogen storage disease type 2 (GSD2) [MIM:232300]; also called acid alpha-glucosidase (GAA) deficiency or acid maltase deficiency (AMD). GSD2 is a metabolic disorder with a broad clinical spectrum. The severe infantile form, or Pompe disease, presents at birth with massive accumulation of glycogen in muscle, heart and liver. Cardiomyopathy and muscular hypotonia are the cardinal features of this form whose life expectancy is less than two years. The juvenile and adult forms present as limb-girdle muscular dystrophy beginning in the lower limbs. Final outcome depends on respiratory muscle failure. Patients with the adult form can be free of clinical symptoms for most of their life but finally develop a slowly progressive myopathy.
  • Sequence similarities

    Belongs to the glycosyl hydrolase 31 family.
    Contains 1 P-type (trefoil) domain.
  • Post-translational
    modifications

    The different forms of acid glucosidase are obtained by proteolytic processing.
    Phosphorylation of mannose residues ensures efficient transport of the enzyme to the lysosomes via the mannose 6-phosphate receptor.
  • Cellular localization

    Lysosome. Lysosome membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • 70 kDa lysosomal alpha-glucosidase antibody
    • Acid alpha glucosidase antibody
    • Acid maltase antibody
    • Aglucosidase alfa antibody
    • Alpha glucosidase antibody
    • GAA antibody
    • Glucosidase alpha acid (Pompe disease glycogen storage disease type II) antibody
    • Glucosidase alpha acid antibody
    • Glucosidase alpha antibody
    • LYAG antibody
    • LYAG_HUMAN antibody
    • Lysosomal alpha glucosidase antibody
    see all

Images

  • Anti-GAA antibody (ab102815) at 1/500 dilution + Mouse intestine lysate at 50 µg

    Predicted band size: 105 kDa

  • All lanes : Anti-GAA antibody (ab102815) at 1/500 dilution

    Lane 1 : GAA transfected 293T cell line
    Lane 2 : Non transfected 293T cell line

    Lysates/proteins at 25 µg per lane.

    Predicted band size: 105 kDa

  • IHC image of GAA staining in human placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab102815, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

ab102815 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

That seems like a good idea.

I would possible also suggest reducing the washing conditions used slightly, using 3x5 minute washes with TBST rather than extended periods.

Please do let me know how you get on.

Read More

Answer

Thank you for contacting us.

I now have some further information in regards to how the western blot presented on the datasheet of Anti-GAA antibody (ab102815) was performed.

5% non-fat milk in PBST (0.2%) was used to block the membrane for 1 hour a room temperature.

The working concentration of the primary antibody was 1ug/mL and theprimary antibody was diluted in fresh blocking buffer [5% non-fat milk/PBST (0.2%)]. The antibody wasincubatedovernight at4℃.

It therefore seems that the blocking and diluent you have been using should not be contributing to the problem. If you could pleaseprovide the information requested inmy previous email hopefully I can try to isolate where the problem may be coming from, and if a replacement antibody may be required.

From my previous email:

I was hoping you may be able to clarify a little further the problems you have been experiencing. Do you observe no bands at all or is there high background? Would it be possible to send an image of the results obtained? I also have a few further questions:

1. how was the tissue samples prepared? How were they lysed and were any protease inhibitors used?

2. Was the sample run denatured or native? Was boiling and reducing agents employed?

3. What buffer was employed in the wash steps?

4. Do you have the order number (or purchase order number) of ab102815?

This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I was hoping you may be able to clarify a little further the problems you have been experiencing. Do you observe no bands at all or is there high background? Would it be possible to send an image of the results obtained? I also have a few further questions:

1. how was the tissue samples prepared? How were they lysed and were any protease inhibitors used?

2. Was the sample run denatured or native? Was boiling and reducing agents employed?

3. Whatbuffer was employed in the wash steps?

4. Do you have the order number (or purchase order number) of ab102815?

This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


I look forward to receiving your reply.

Read More

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