Overview

  • Product name

    Anti-GAD65 + GAD67 antibody
    See all GAD65 + GAD67 primary antibodies
  • Description

    Rabbit polyclonal to GAD65 + GAD67
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, IHC-Fr, WB, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Chicken, Cat, Human, Zebrafish, Macaque monkey
    Predicted to work with: Pig
  • Immunogen

    Synthetic peptide:

    DFLIEEIERLGQDL

    , corresponding to C terminal amino acids 572-585 of Human GAD65+67.

  • Positive control

    • Recombinant Human GAD67 protein (ab114255) can be used as a positive control in WB. Brain.
  • General notes

    This product should be stored undiluted. Storage in frost free freezers is not recommended. Should this product contain a precipitate we recommend microcentrifugation before use. Shelf Life: 12 months from date of despatch.

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

    Preservative: 0.09% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity

    IgG fraction
  • Purification notes

    An immunoglobulin-rich fraction was prepared from whole serum by ammonium sulphate precipitation.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab11070 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 Use at an assay dependent concentration. PubMed: 20650017
IHC-Fr 1/200 - 1/5000.
WB 1/1000 - 1/10000. Predicted molecular weight: 65 , 67 kDa.
ELISA Use at an assay dependent concentration.

Target

Images

  • ab11070 staining GABA transporter 2 in zebrafish retina sections by IHC-Fr. The tissue was fixed with paraformaldehyde and an antigen retrieval step was performed with sodium citrate pH 6. Blocking of the sample was done with 5% BSA in PBS containing 0.5% Triton X, for 60 minutes at 23°C, followed by staining with ab11070 at 1/200 in blocking solution for 16h at 4°C. An alexa 488 conjugated goat anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei are stained in blue with DAPI. GAD65/67 expression can be observed in amacrine cells (in green).

    See Abreview

References

This product has been referenced in:

  • Stevenson SA  et al. Intervention changes acoustic peak frequency and mesolimbic neurochemistry in the Pink1-/- rat model of Parkinson disease. PLoS One 14:e0220734 (2019). Read more (PubMed: 31374106) »
  • Samarut É  et al. ?-Aminobutyric acid receptor alpha 1 subunit loss of function causes genetic generalized epilepsy by impairing inhibitory network neurodevelopment. Epilepsia 59:2061-2074 (2018). Read more (PubMed: 30324621) »
See all 35 Publications for this product

Customer reviews and Q&As

1-8 of 8 Q&A

Answer

Thank you for your reply. I recommend the following HRP conjugated secondary antibodies to use with these rabbit Anti-GAD65+GAD67 primary antibodies. I have included one product from a donkey host and another from goat. Both are guaranteed and the choice will be personal preference of host. These products are below. Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) Ab16284: https://www.abcam.com/Rabbit-IgG-secondary-antibody-ab16284.html Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) Ab97051: https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-HL-HRP-ab97051.html Abcam also provides a number of IHC reagents and kits such as DAB substrate kits, donkey or goat sera and antigen retrieval buffers. All of our products are covered by our six month Abpromise guarantee and are fully supported by our scientific support staff. I have provided links to a few of these products below. DAB substrate kit:https://www.abcam.com/DAB-substrate-kit-ab94665.html Donkey serum:https://www.abcam.com/Donkey-Serum-ab7475.html Goat serum:https://www.abcam.com/Goat-Serum-ab7481.html Antigen retrieval buffer:https://www.abcam.com/Antigen-Retrieval-Buffer-100X-Tris-EDTA-Buffer-pH-9-0-ab93684.html I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.

We are happy to help customers find the most suitable products for their research purposes; however we are not specialized in this particular field and do not want to recommend a product that may be unsuitable. We encourage customers to consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about their specific research interests.

All three of ourAnti-GAD65 + GAD67 antibodies are guaranteed to work in mouse tissues. Ab49832 and Ab11070 are both tested and guaranteed in IHC. I would be very happy to help you to find the best secondary antibody for use with either of these products. In order to determine this could you please let me know the detection system you plan to use?

I am sorry that I could not be more helpful regarding the best marker for your needs, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

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Answer

Gracias por contactarnos de nuevo.

Siento mucho los problemas ocasionados por estos productos que no han funcionado como se esperaba.

Para poder dar el soporte oportuno de los mismos y entender por qué han dado ese resultado, te rogaría que por favor me ampliaras la siguiente información:

- ¿Cuál es el numero de pedido de los anticuerpos?

- ¿Cuáles son los lotes que funcionaron en el pasado de ab59791 y ab49917?

- ¿En qué condiciones se ha llevado a cabo el lisado de tejidos (buffer, inhibidores, agente reductor, etc...)?

- ¿Se ha usado un control positivo en paralelo con las muestras?

- ¿Se ha comprobado el buen estado del secundario utilizado, así como del sustrato DAB? Es decir, ¿se han usado con otros sistemas obteniendo un resultado positivo?

- Recomendamos llevar a cabo una incubación de anticuerpo de 12-16 horas para asegurar que el anticuerpo alcanza el epítopo.

Por otro lado, quisiera asegurar al cliente que los anticuerpos no han podido afectarse por el hecho de llegarles a 8C. El envío del producto en los bloques azules de hielo es solo una medida de precaución, y ésta no es la primera vez que un cliente nos reporta la llegada de los productos a temperatura ambiente, sin que posteriormente haya sufrido algún problema con el mismo.

En nuestros laboratorios hemos hecho estudios de estabilidad de los anticuerpos y proteínas con la temperatura, demostrando que son estables hasta a 45C durante una semana sin que pueda impactar en su actividad.

El lunes estaré fuera de la oficina y no podre contestar los correos, pero a partir del martes día 2 podre segur adelante con esta reclamación. Espero tu respuesta entonces.

Que pases un buen día.

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Answer

Thank you for your reply. I hope the increased antibody will help to label the cell bodies. If you are able to get specific signal, then you may be able to reduce the background by shortening your antigen retrieval or increasing the blocking time. If not, please let me know your original order or PO number and I will be happy to help you further.

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Answer

Thank you for contacting us. I am sorry that this antibody has been giving no signal in your IHC. Could you please clarify a few points of your protocol?

1) What type of samples are you working with? What species and tissue type? How long were your samples fixed for?

2) Did you observe any background signal or were the sections completely blank? Have you used your secondary antibody recently with other primary antibodies? What secondary antibody were you using?

3)In some cases, long wash steps can reduce the antibody binding. I would recommend trying 3 x 5 minute washes after the primary and secondary antibody incubations.

4)Depending on the expression of GAD65 + GAD 67 in your samples, you may need to use up to a 1:200 dilution for this primary antibody. Using more antibody may help to increase your signal.

I hope this helps, if not, please let me know your original order or PO number and I will be happy to assist you further.

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Answer

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products. I would like to confirm that you are eligible for our testing discount program if the antibody has not yet been purchased. If you are interested I will issue an 100% discount code for you. You would have to buy ab11070 and then test ab11070 with the mentioned recombinant proteins in WB. After submitting an Abreview the discount could would become active and you would be able to order your free antibody as an reward for testing this for future users and us at Abcam. To find out more about our Abreview system, please see the following link: https://www.abcam.com/abreviews I hope this information is helpful. Please do not hesitate to contact us again with any other questions or if your are interested in this offer.  

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Answer

Thank you for participating in our testing discount program. When you submit your abreivew, please be sure to note the testing discount code in the "additional notes" section. The website does have specific questions for you to answer as well as the notes section for any additional information you may have. Once you have submitted the review you will be able to use the code for an additional antibody. You will need to specify the code to the customer service agent when ordering so that you receive the antibody free. The testing discount program only provides a free antibody per each purchased antibody, so you would not be able to receive another code for this free antibody. Please let me know if you have any further questions.

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Question

ANTIBODY CODE ab11070 DESCRIPTION OF THE PROBLEM Multiple bands, wrong band size. SAMPLE plant cell extract from plants transformed to express GAD65 PRIMARY ANTIBODY antiGAD65+67 (ab11070), abcam SECONDARY ANTIBODY antirabbit Ig, 1:1000, in PBSTM (1 % milk powder), 1 h at room temperature, 3x5 min washing with PBSTM DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Positive control: human brain normal tissue lysate (ab7918) Negative control: tobacco wilt-type plants extract ANTIBODY STORAGE CONDITIONS -20 C ab diluted in PBSTM (milk powder 5%) SAMPLE PREPARATION Hepes, PMSF, DTT, sodium metabisulfite, 5 min at 96 C AMOUNT OF PROTEIN LOADED 20 ug ELECTROPHORESIS/GEL CONDITIONS 7% stackin gel, 15% resolving gel, Lammlei buffer, 70 mA TRANSFER AND BLOCKING CONDITIONS 5% milk powder in PBST overnight at 4 C HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 6 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? increased milk powder concentrario up to 10% in PBST tried dilutions of primary ab of 1:5000, 1:10000, 1:15000 and 1:20000 tried preabsorbing antibody with wild-type extract (acetone powders) ADDITIONAL NOTES In the positive control I get a band of the expected size (65 kDa) and another one of about 20 kDa. When detected with antiGAD65+67, brain lysate should show two band, one at 65 and one at 67 kDa. The band at 65 is not present in the wild type nor in the transgenic lines. However, there's a band of 20 kDa that is present in the transgenic lines and in the positive control (even though this band is not expected) but not in the wild-type. I've checked the presence of the gene by PCR in the transgenic lines and it is apparently present.

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Answer

Thank you for your enquiry and for submitting more details of your protocol. We are very sorry to hear that you are having problem with this antibody. We have searched our database and found that this is a popular selling product and your feedback is the first we have received about it not working. Therefore, at this stage, we would suggest that there is either a problem with the vial you received, or modifications to your protocol are needed to obtain a positive result. Looking at the information supplied above, it would appear that you have diluted the antibody prior to storage. This is not recommended and therefore may have caused a detrimental effect to the antibody.

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