Overview

  • Product name
    Anti-Quinolinic acid antibody
  • Description
    Rabbit polyclonal to Quinolinic acid
  • Host species
    Rabbit
  • Specificity
    This antibody targets conjugated quinolinic acid, it doesn't recognize free quinolinic acid.
  • Tested applications
    Suitable for: IHC-Fr, IHC-Pmore details
  • Immunogen

    Chemical/ Small Molecule corresponding to Quinolinic acid conjugated to Bovine Serum Albumin (BSA).

Properties

Applications

Our Abpromise guarantee covers the use of ab37106 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-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. PubMed: 21831269

Target

  • Relevance
    Quinolinic acid is a naturally occurring toxin in the central nervous system believed to play a role in AIDS dementia complex. A metabolite of tryptophan with a possible role in neurodegenerative disorders. Elevated csf levels of quinolinic acid are significantly correlated with the severity of neuropsychological deficits in patients who have aids.
  • Alternative names
    • Pyridine 23 dicarboxylic Acid antibody

References

This product has been referenced in:
  • Busse M  et al. Decreased quinolinic acid in the hippocampus of depressive patients: evidence for local anti-inflammatory and neuroprotective responses? Eur Arch Psychiatry Clin Neurosci 265:321-9 (2015). Read more (PubMed: 25409655) »
  • Steiner J  et al. Severe depression is associated with increased microglial quinolinic acid in subregions of the anterior cingulate gyrus: evidence for an immune-modulated glutamatergic neurotransmission? J Neuroinflammation 8:94 (2011). IHC-P ; Human . Read more (PubMed: 21831269) »
See all 2 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you very much for your interest in ab85518 and ab37106.

To our knowledge, ab85518 and ab37106 have not been tested in flow cytometry.If however you would like to test this,I can offer a discount off a future purchase if you buy ab85518 or ab37106 now, test it inflow cytometryand submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab85518 or ab37106 in flow cytometry. I will then send a discount code. This code must be issued before purchasing ab85518 or ab37106 so please wait for my reply before ordering.

2. Purchase ab85518 or ab37106 either by phone, fax, or online (www.abcam.com).

3. Test it in flow cytometry.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab85518 or ab37106 turns out to be unsuitable for flow cytometry, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer



I don't think it will be possible to detect free quinolinic acid by ELISA using ab37106 and ab88518 in a sELISA. These antibodies will not detect free quinolinic acid and they have never been tested together in a sELISA so their epitopes are not known to be compatible. The ELISA protocol that was used with each antibody was to detect the quinolinic acid conjugated to BSA (which we carry as ab45209) in an indirect ELISA assay. The protocol is below:

1. Coat maxisorp well plates (Nunc) with a solution of antigen (the dilutions for antigen range from 4-50 μg/ml, please see ab45209) in sodium carbonate buffer 0.05 M (pH 9.6) containing sodium metabisulfite (SMB) (Acros) 0.001 M, for sixteen hours at 4°C.

2. Saturate well plates with a solution of PBS (pH 7.3) containing 2.5 g/l of BSA (Acros), 0.05% Tween 20 (Acros) and SMB 0.001 M for one hour at 37°C.

3. Wash with PBS/0.5% Tween three times.

4. Dilute primary antibody as per antibody data sheet in PBS containing 2.5 g/l BSA, 10% of glycerol and SMB 0.001 M, 200 μl by well plate (incubating for 2 hours at 37°C).

5 . Wash with PBS/0.5% Tween three times.

6. 200 μl of peroxidase-labeled goat anti- "primary antibody host species" IgG diluted (1/10,000) in a solution of PBS containing 2.5 g/l BSA, 10% of glycerol, 0.5% Tween and SMB 0.001 M, will be applied by well plate (for one hour at 37°C).

7. Rinse well plates with PBS/0.5% Tween three times.

8. And finally, develop the peroxidase by incubating 200 μl by well plate of a citrate 0.1 M/phosphate, 0.2 M (pH 5) solution containing 0.4% of OPD (Sigma) and 0.03% of hydrogen peroxide (Acros) for ten minutes in the dark. After that, stop the reaction by the addition of 50 μl of 2 M HCl.

9. The optical density will be measured at 492 nm, to obtain the different values (IC50).

Unfortunately we do not have any products capable of detecting free quinolinic acid. I took a look at the literature, and as you probably already know it actually is quite involved (usually H NMR, HPLC, etc.).

http://www.cyberneum.de/fileadmin/user_upload/files/publications/pdf2041.pdf

http://www.sciencedirect.com/science/article/pii/0003267094004936

http://www.sciencedirect.com/science/article/pii/S0021967313015793

I am really sorry that these products are not designed to do the experiments you were hoping to run, but please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

These antibodies will not recognize free quinolinic acid. They will only recognize the quinolinic acid in the presence of gluteraldehyde, either as part of the protein carrier linkage, or as a component in the tissue profusion step.

Perfusion protocol for Adult male Sprague Dawley (weight around 0.5 kg) :

1. The animals can be deeply anaesthetized, for example with urethane (0.5-1.5g/kg, intraperitoneal).

2. Heparinize and perfuse via the ascending aorta with 100 ml of cold physiologic saline (0.9% NaCl)

and with the following fixative solution:

a) 300 ml of cold 4% paraformaldehyde and 2% glutaraldehyde in 0.1 M phosphate-buffer (PB), pH

7.2 (two minutes).

b) 600 ml of cold 4% paraformaldehyde and 2% glutaraldehyde in 0.1 M PB, pH 7.2 (ten minutes).

c) Dissect out the brains and place in a solution of 4% paraformaldehyde in 0.1 M PB, pH 7.2, at 4°C

for twelve to sixteen hours.

d) Before the brains are cut on a freezing microtome, infuse the brain in increasing concentrations of

sucrose (a first bath of 5% of sucrose in PBS until the brains sink), after that repeat the same process

in a solution with a higher level of sucrose (10%, 20%, 25% and finally 30%).

3. Cut serial sections around 50 μm-thick. Keep at 4°C in PBS (0.1 M, pH 7.2) and process for

immunostaining.

Immunostaining protocol:

1. In order to avoid possible interference with endogenous peroxidase, treat free-floating sections with

distilled water containing NH3 (20%), H2O2 (30%) and NaOH (1%) for 20 minutes (other method is

using a solution with 33% of H2O2 and 66% of methanol).

2. Wash the sections for 20 minutes in 0.15 M phosphate-buffered saline (PBS) (pH 7.2).

3. Pre-incubate for 30 minutes in PBS containing 10% of normal horse serum and 0.3% of Triton

X-100 (mixed solution).

4. Incubate at room temperature (1 hour 30 minutes) and overnight at 4°C mixing with a solution

containing primary antibodies.

5. Wash for 30 minutes with PBS.

6. Incubate for 60 minutes at room temperature with biotinylated anti-rabbit IgG (Vector) diluted 1/200

in PBS.

7. Wash for 30 minutes with PBS.

8. Incubate sections for 1 hour with a 1/100 diluted avidin-biotin-peroxidase complex (Vectastain).

9. Wash for 30 minutes with PBS.

10. Wash with Tris-HCl buffer (pH 7.6)(10 minutes).

11. Develop the tissue-bound peroxidase with H2O2 using 3, 3' diaminobenzidine as chromogen.

12. Rinse the sections with PBS and coverslip with PBS/Glycerol (1/1).

They don't have a specific WB protocol, but they do have the antigen (attached to BSA) that could be used as a positive control.

The linkage between Quinolinic acid and BSA is by 1-(3-Dimethyl-aminopropyl)-3 ethyl carbodiimide. The conjugation site is one or both of the carboxyl groups on quinolinic acid.

For ab37106, here is another reference: Guillemin GJ, Brew BJ, Noonan CE, Takikawa O, Cullen KM (2005) Indoleamine 2,3 dioxygenase and quinolinic acid immunoreactivity in Alzheimer's disease hippocampus. Neuropathol Appl Neurobiol 31(4):395-404.

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Answer



I am happy to confirm that both antibodies are recognizing t to their target in dependent of the species that sample originates from.

The targets are not proteins or peptides but are chemical compounds (metabolite) that have the same structure in every species.

Therefore I can recommend both antibodies for your experiment with rat samples. Please make sure to check if your planned application is tested.

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Answer


Using a quinolinic acid adsorbed on bovine serum albumin, antibody specificity was performed with an ELISA test by competition experiments with the following compounds:

Compound Cross reactivity*

Quinolinic acid-BSA 1
Kynurenic-BSA 1/50,000
Phenylalanine-BSA 1/50,000

* Quinolinic acid-BSA concentration/unconjugated or conjugated concentration at half displacement.

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Answer

Thank you for your inquiry.

I can confirm that ab37106 is tested and guaranteed for ELISA, IHC-Fr (PFA fixed)andIHC-P.

All information we have we state on the datasheet.

Unfortunately, we do not currently have more information about this product. We will update the on-line datasheet for this product as soon as we receive additional data or images.

We apologize for any inconvenience this may cause.

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Answer

Thank you for your enquiry.

I can confirm that ab37106Quinolinic acid antibody is sold as whole antiserum. Unpurified antibodies, such as those sole as whole antiserum, will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, andserum will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for whole antiserum, concentration of antibody is known to very between 1- 10 mg/ml.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this informationwill be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Answer

Thank you for your enquiry. I am afraid that we do not know the exact concentration of this product since unpurified antibody preparations vary significantly in specific antibody concentration. However, there are some “typical ranges”, which can be used as a guideline for estimation. In this case, the concentration may be estimated to be around 10-16mg/ml. I hope this information will be helpful to you. Should you require any further assistance, please do not hesitate to contact me.

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

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