Overview

  • Product name
    Anti-Glutamine Synthetase antibody
    See all Glutamine Synthetase primary antibodies
  • Description
    Rabbit polyclonal to Glutamine Synthetase
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Monkey
    Predicted to work with: Chicken, Hamster, Cow, Dog, Human, Pig, Cynomolgus monkey
  • Immunogen

    Synthetic peptide:

    RTCLLNETGDEPFQYKN

    , corresponding to C terminal amino acids 357-373 of Mouse Glutamine Synthetase

  • Positive control
    • Rat brain tissue, rat brain cytosolic fraction extract, cerebellum, kidney

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: 0.0268% PBS
  • Concentration information loading...
  • Purity
    Ion Exchange Chromatography
  • Purification notes
    Whole antiserum is fractionated and then further purified by ion-exchange chromatography to provide the IgG fraction of antiserum that is essentially free of other rabbit serum proteins.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab49873 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 1/10000. Predicted molecular weight: 42 kDa.
IHC-P 1/10000 - 1/20000.
ICC/IF Use at an assay dependent concentration.
IHC-Fr 1/5000.

Target

  • Function
    This enzyme has 2 functions: it catalyzes the production of glutamine and 4-aminobutanoate (gamma-aminobutyric acid, GABA), the latter in a pyridoxal phosphate-independent manner (By similarity). Essential for proliferation of fetal skin fibroblasts.
  • Involvement in disease
    Defects in GLUL are the cause of congenital systemic glutamine deficiency (CSGD) [MIM:610015]. CSGD is a rare developmental disorder with severe brain malformation resulting in multi-organ failure and neonatal death. Glutamine is largely absent from affected patients serum, urine and cerebrospinal fluid.
  • Sequence similarities
    Belongs to the glutamine synthetase family.
  • Developmental stage
    Expressed during early fetal stages.
  • Cellular localization
    Cytoplasm. Mitochondrion.
  • Information by UniProt
  • Database links
  • Alternative names
    • cell proliferation-inducing protein 59 antibody
    • GLNA antibody
    • GLNA_HUMAN antibody
    • GLNS antibody
    • GLUL antibody
    • Glutamate ammonia ligase antibody
    • Glutamate decarboxylase antibody
    • Glutamate--ammonia ligase antibody
    • glutamine synthase antibody
    • Glutamine synthetase antibody
    • GS antibody
    • PIG 43 antibody
    • PIG 59 antibody
    • PIG43 antibody
    • PIG59 antibody
    • Proliferation inducing protein 43 antibody
    see all

Images

  • All lanes :

    Lane 2 : 10000
    Lane 3 : 20000

    Secondary
    All lanes : Goat Anti-Rabbit IgG-Peroxidase

    Predicted band size: 42 kDa

  • ab49873 at 1/5000 staining cultured mouse astroglial cells by ICC/IF. The cells were paraformaldehyde fixed and permeabilized with TritonX100 before being blocked and stained with the antibody for 24 hours at 4°C. An Alexa-Fluor ® 594 conjugated chicken anti-rabbit antibody was used as the secondary.

    See Abreview

  • Glutamine Synthetase antibody (ab49873) immunohistochemistry on rat brain and kidney tissue (formalin/PFA-fixed paraffin-embedded) sections. Rat cerebellum and rat kidney sections were incubated with ab49873 (1/21000) for 2h at RT. Antigen retrieval was performed by heat induction in citrate buffer pH6. The upper panel shows immunoreactivity in rat kidney, apart from convoluted tubule positivity (proximal: see asterisks) there is positivity within the glomerular capsule (perhaps the parietal epithelium: see arrows). In the lower image (cerebellum), apart from astrocytic glial cell body and process positivity, I note that what I think are oligodendrocytes in the grey matter, are also positive. In the corpus callosum of the same section (and all other major axonal tracks of this brain), linearly arranged cells are also positive: these are known to be oligodendrocytes.
    .

    See Abreview

References

This product has been referenced in:
  • Afroz S  et al. CGRP Induces Differential Regulation of Cytokines from Satellite Glial Cells in Trigeminal Ganglia and Orofacial Nociception. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30736422) »
  • Zhou M  et al. Non-cell-autonomous activation of IL-6/STAT3 signaling mediates FGF19-driven hepatocarcinogenesis. Nat Commun 8:15433 (2017). IHC-P ; Mouse . Read more (PubMed: 28508871) »
See all 21 Publications for this product

Customer reviews and Q&As

11-19 of 19 Abreviews or Q&A

Answer

Thank you for getting back to me.

I am sorry that the reply to your fax was sent to your supervisor by one of my colleagues, Keith. This was due to the way our assigning system works. The protocol tips that I gave and those of Keith are not that dissimilar. We both recommended optimising the dilution of the antibody. Keith also recommended using Glycine to reduce the possible background created from uncapped aldehyde groups. I did not mention this as you were having problems seeing any staining, the background was therefore not my top priority to tackle. What may be significantly contributing to your lack of signal may be the way that the tissue is fixed. Over fixation can lead tothe epitopeto be probed to bemasked.Even with permiabilisation, thismay not be unmasked. Someform of antigen retrieval may therefore be necessary.In my previous email I tried to find out a little more about this in order to be able to provide advice on how this could be improved:

1. Exactly how was the tissue fixed? For how long was the tissue treated with PFA? What concentration of PFA was used? At what temperature?

2. Has the goat anti-rabbit secondary antibody used for both experiments been used successfully before?

3. With the tissue sections that you have, have they been successfully stained to detect a different target with a different antibody? Could you send me some images of this staining?

If you could also let me know under exactly what conditions the new experiment was carriedout, that would be helpful. Which ofmy suggestionsdid you try?

As I mentioned in my previous email, ab93434 has to our knowledge not been used in staining of frozen sections and it may be that the antibody is not suitable for this application. As this application is not listed on the datasheet the antibody is therefore not covered by the Abpromise for this application.

As you are having problems with both of these very different antibodies and I am as yet not sure what is contributing to this problem, it seems that it may either be the tissue being used or the protocol which may be contributing to the lack of signal. As I want you to get the best staining that you can, I would therefore like to ascertain if it is a problem of the antibodies first. Have you performed a western blotting experiment with these antibodies? If not, I would like to provide you with a positive control for this application to test the activity of the antibodies.

If you would like for me to send this positive control for western blotting please do let me know. Otherwise, all I can offerat this time is a refund, in addition to the ab91073 which I offered as a good-will gesture.

I hope this has been of help. I look forward to receivingyour reply.

Read More

Answer

Thank you for providing me with this extra information. It has allowed me to more fully understand the problems you have been experiencing with ab49873 and ab93434 previously.

Having looked at the protocol I have a few further questions:

1. Exactly how was the tissue fixed? For how long was the tissue treated with PFA? What concentration of PFA was used?

2. What were the samples tested? Which species did the tissue come from?

3. Had the goat anti rabbit used for both of the experiments been used successfully before?

As ab93434 has as yet to our knowledge not been used in staining frozen sections it may be that the antibody is not suitable for this application. However, there are a few suggestions I can make which may help toimprove the results seen so far.

1. I would suggest decreasing the dilution of both the antibodies. Although ab49873 is recommended to be used at 1/5000, this is only a starting point and may need to be optimised depending on the tissue and protocol used. With ab49873, I would suggest initially trying 1/500 to see if there is any signal. With ab93434 we recommend using 5 µg/ml as an initial starting point, I would therefore use a dilution of 1/50 to start.

2. As you have not been observing any staining with overnight incubation at 4 degrees, I would try incubating at room temperature for 2 hours to see if this improves the results.

3. I would increase the level of Triton X used to 0.2 % for performing the permiabilisation.

I hope these protocol tips are of use, with the answers to the questions above I may be able to provide further suggestions.

I am able to send you ab91073 directly as I suggested to detect theL isoform of glutaminase as a good-will gesture for the bad experience you have had. If you would prefer to change this to ab60709 I can also organise this.

I look forward to receiving your reply.

Read More

Question
Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab49873 with the order number 972573. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Monkey Tissue sections (Cerebellum)
Specification
Cerebellum
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C

Mr. Carl Hobbs

Verified customer

Submitted Apr 21 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Brain cortex and colon)
Specification
Brain cortex and colon
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C

Mr. Carl Hobbs

Verified customer

Submitted Jan 22 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney and cerebellum)
Specification
Kidney and cerebellum
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid 10mM pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C

Mr. Carl Hobbs

Verified customer

Submitted Apr 01 2009

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Specification
Brain
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C

Mr. Carl Hobbs

Verified customer

Submitted Feb 09 2009

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (early postnatal brain section)
Specification
early postnatal brain section
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 16 2008

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cultured Cells (astroglia)
Specification
astroglia
Fixative
Paraformaldehyde
Permeabilization
Yes - TritonX100 0.1%
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. Zsuzsanna Kornyei

Verified customer

Submitted Oct 29 2007

11-19 of 19 Abreviews or Q&A

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