Anti-gamma Synuclein antibody (ab6169)

Thank you and your customer for taking the time to complete our questionnaire and contact us. I am sorry to hear your customer has had difficulty obtaining satisfactory results from this antibody.

The details your customer has kindly provided will enable us to investigate this case for your customer and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab6169:

1.) I suggest to use positive control. Wehave positive data with MCF7 cells. I could not find any evidence that gamma Synuclein is expressed in rat tendons or fibroblasts.
It is described as specifically expressed in the peripheral nervous system with high expression in motoneurons of the brainstem and it lso found in neurons of many other brain regions including the cerebellar cortex, thalmus, hypothalamus and CA1, CA2, CA3 and CA4 regions of the hippocampus.

http://www.uniprot.org/uniprot/Q63544

http://biogps.org/#goto=genereport&id=6623(choose species rat)

2.) General suggestion would include to incubate the primary antibody over night at 4C and to use BSA as blocking agent. This would ensure a stronger signal.

We are happy to offer this technical support. In the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I hope this information is helpful, thank you and your customer for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again.

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. 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 with the order number xxxx. 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.

Thank you for contacting Abcam. At Abcam, providing quality service is just as important as providing quality products. Our Abpromise® ensures that you can trust our products to perform as stated on the datasheets, or we will offer a replacement, credit, or refund. For our records, could you please send me the protocol that you have used with ab47966. Including cell preparation, blocking solutions, times and temperature as well as antibody incubation times and any controls that you may have used. If possible, would you be able find the lot number of the previous vial or an order number from that purchase as well? The Abcam Abpromise® guarantee: • 100% Scientific and Customer Support for any product you buy from Abcam or one of our authorized distributors. • We guarantee our products work in the tested species and applications stated on the datasheet. • We will replace or refund products not performing as stated on the datasheet if reported within 6 months of purchase. • We investigate any quality concerns raised by customers to ensure our catalog contains products that perform to the highest standards. Please note these conditions to our Abpromise®: • Protocol information must be provided in order for the claim to be validated. • Antibodies tested in recombinant samples only are not guaranteed for use on endogenous samples. • “Predicted to react” information on the datasheets is provided for reference only and these species are not guaranteed. • Fast Track antibodies are covered for ELISA against the immunizing peptide only. I look forward to hearing back form you and working through this issue with you. I hope this information is helpful, but please do not hesitate to contact us with further questions.

Thank you for your enquiry. The only gamma-synuclein antibody which we currently have is ab6169, which is not directed against mouse. To see if another company may have what you are looking for, I suggest that you click on the link to "The World's Antibody Gateway". The World's Antibody Gateway is a free search engine service provided by Abcam to help you to quickly find the antibodies that you are looking for. It is a free-text search engine developed by the Abcam team so that it searches the catalogs of all online antibody companies (currently 249).

We tested the antibody on recombinant protein only 100ng recombinant protein each lane. The primary was 1/2000, secondary was alkaline phosphatase conjugated anti-rabbit from Sigma. We did not try it on crude samples such as brain or other tissue homogenates. I have attached the Western blot image to the datasheet so please refer to that.

The antibody should work on paraffin embedded breast cancer tissue, as it works fine with paraffin embedded human brain tissue which contained abundant gamma-synuclein. However, we have not tried any non-neural tissues.

For neurological tissue, the use of a scientific microwave is preferable, as it can be set to just below boiling to help minimise the risk of section dissociation. We have no experience of this particular antibody in non-neural tissue. For a general antigen retrieval and IHC protocol on formalin fixed paraffin embedded tissue, please see the links on the right hand side of the Abcam homepage.

Try using this antibody at the recommended dilution of 1:1000, then adjust the concentration accordingly from the results. Make sure you perform antigen retrieval before staining (I'll send you my protocol). If doing immunofluorescence, use PBS for buffer washes and to make up reagents. If doing immunoperoxidase or phosphatase use TBS with 0.025% Triton for buffer washes (except after the application of the secondary, use TBS alone). If you're not using a kit for immunoperoxidase or phosphatase, make all reagents up in TBS with 1% BSA. (Leave the BSA out of the ABC, if you're using it, and the chromagen). Use a humidified incubation chamber to avoid slides drying out. Block in normal rabbit serum for 2 hours before adding the primary. Use at 1:10 Incubate the primary at 4 degrees centigrade over night. If using HRP, block endogenoue peroxidases with hydrogen peroxide after adition of the secondary. Likewise if using AP, but use Levamisole. If using AEC as a chromagen, use an aqueous mountant and do not dehydrate through alcohol. I hope that these tips are of use to you.

If the centriole contains tubulin then this antibody will detect it.