Anti-Semaphorin 3A antibody (ab23393)

Hello and thank you for contacting us. This antibody have not been tested for sandwich ELISA with a paired antibody, nor has the detection limit of the antibody been tested. I am sorry I could not be more helpful in this case. Please let me know if you have any further questions.

Thank you for your reply.
My colleague is currently out of the office and I'm afraid my Spanish is not good enough to reply in this language.
Unfortunately testing customers samples in our lab is not possible.
We appreciate the time you have spent on these experiments and, as agreed, we are pleased to arrange a replacement with one unit of ab23393. The order number is: *****.
To check the status of the order please contact our Customer Service team and reference this number.
About the lysis buffer, I would recommend to use a RIPA buffer (Radio Immuno Precipitation Assay buffer):
150 mM sodium chloride
1.0% NP-40 or Triton X-100
0.5% sodium deoxycholate
0.1% SDS (sodium dodecyl sulphate)
50 mM Tris, pH 8.0

For more details about the choice of lysis buffer and Western Blot I would suggest to look at our recommended protocol available on our website:
Protocols homepage: https://www.abcam.com/index.html?pageconfig=popular_protocols
Western Blot: https://www.abcam.com/index.html?pageconfig=resource&rid=11375
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.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Ya me han llegado las imágenes, muchas gracias por haber insistido.
Me confirman que este anticuerpo se ha usado en WB con lisados de cerebro de ratón a diluciones 1/250 y 1/1000. La incubación se ha llevado a cabo durante 2 horas a temperatura ambiente y quimioluminiscencia como sistema de detección.
El péptido inmunógeno está disponible en nuestro catálogo:
https://www.abcam.com/Semaphorin-3A-peptide-ab42768.html
y si te interesa te lo puedo enviar de forma gratuita para que lo probéis, confiando en que os ayude a evaluar el resultado obtenido.
Espero tu respuesta al respecto, y confío en que esta información sea de utilidad.

Thank you for your call today and for your patience.

I have discussedthe purification kit ab102783with my colleagues, and the minimum amount of antibody that is recommended to use with the kit is 20 ug.Since there are 50 ug per vial of ab23393, this kit will be compatible, and youcan dilute the antibody to 100 uL(the minimum volume required).

The protein A usedwiththis kit has a high affinity for rabbit IgG, so the expected recovery is greater than 95%. The IgG will be edulted in 200-300 uL of buffer, however, so it will be more dilute than the original concentration. The neutralisation buffer is not compatible with most BCA or Bradford reagents, so the purified concentration should be measured before addition of the neutraliser, or an OD280 should be used to measure concentration.

I hope this information will be useful, but if you have any further questions or if there is anything else that we can do for you, please let me know and I'll be happy to help.

Thank you for your call today, and I apologize for the trouble with these codes! I have created new codes which will expire on March 1, 2012. I will go through your Abreviews and put these new codes in our internal notes so that the discounts can be applied to your orders tomorrow. Please let me know if you have any questions, or if there is anything else that we can do for you.