Anti-ATG8 抗体 (ab4753)

As far as we are aware, ab4753 has never been tested with samples from P.pastoris. All tested species by our 6 month guarantee are stated on our datasheets, and these are updated as soon as any new information is brought to our attention.

I am sorry I have not been able to find the the P pastoris ATG8 amino acid sequence on the protein database sites in order to check the alignment with the immunogen. If you have the sequence available, I would be pleased to check this for you.

If the alignment is high, this will indicate the antibody is likely to detect in P pastoris. If you would like to test the antibody in P pastoris, please do not hesitate to contact me with the sequence prior to the purchase as you may be eligible for our AbTrial testing discount program. <

This antibody will primarily detect one band at 14 kDa, but may also recognize other prominent intrinsic bands corresponding to UBLs or their conjugates.

Could you send us the protein sequences specific to species in question?We can then check for homology.

Thank you very much for contacting us with your question.
I ran an alignment between the immunogen of ab4753 (full-length S. cerevisiae protein) and the C. reinhardtii autophagy protein sequence, and there is 75% identity between the sequences. It is possible that the antibody will be cross-reactive with Chlamydomonas, but we can't say for sure.
We don't offer small samples for testing, but we do have a testing discount program if you'd still like to try this antibody. As part of this program, you would first purchase and test ab4753, and then submit an Abreview with your data and protocol. In exchange we will give you a $347 discount (the price of ab4753) on a future purchase.
I hope that this information will be useful, but if you have further questions or need anything else, please let me know and I'll be happy to help.

Muchas gracias por tu respuesta y por enviar las imágenes.

Efectivamente es posible que el vial concreto que hemos enviado al cliente se haya degradado. Dado que el producto está bajo garantía, estaré encantada de reemplazarle o reembolsarle su importe.

El único lote actualmente disponible de este anticuerpo es el mismo que recibió el cliente en el último pedido, me temo que no hay más en stock.

Espero tu respuesta para que me indiques cómo proceder y poder ayudar al cliente.

Gracias por contactarnos y mandarnos el cuestionario completo. Lamentablemente no he recibido la imagen adjunta en el cuestionario, por lo que agradecería que si es posible la volvierais a enviar.

En la datasheet del anticuerpo se indica que éste reconoce otras bandas intensas correspondientes a derivados de ubiquitinas.

Sin embargo, atendiendo al protocolo seguido, hay una serie de sugerencias que posiblemente mejoraran el rendimiento del anticuerpo, disminuyendo el ruido de fondo:

- Usar inhibidores enzimáticos, especialmente inhibidores de proteasas, evitan la degradación de la proteína.

-Para aumentar las condiciones reductoras yo sugeriría aumentar el tiempo de hervido de las muestras a 10 minutos.

-La dilución usada de anticuerpo es muy baja. Nosotros recomendamos usar diluciones de entre 1/4000- 1/8000. Igualmente, aumentar la dilución del secundario (por ejemplo a 1/6000) puede disminuir el background.

-Otra sugerencia es cambiar las condiciones de bloqueo. Usando BSA al 5% durante 2 horas.

Confió en que estas recomendaciones mejoren el resultado. En caso contrario, por favor, no dudéis en contactarme de nuevo.

Thank you for contacting us. Unfortunately, I was unable to view the image provided in the word document. Is is possible for you to send a copy of the image separately?

Based on the customer's protocol, it may be possible to obtain more specific results using less primary antibody. We typically recommend using a 1:4000 dilution. It is also possible that the customer's prolonged washes are removing the blocking agent from the membrane and causing background. For that reason I would recommend performing 3 x 5 minute washes.

Regarding the lack of signal, it may be due to the lysis in the 100% trichloroacetic acid. Using a gentler lysis buffer such as a 1% NP-40 or 1% Triton X-100 in TBS may help to better preserve the protein epitopes.

I hope this helps to improve the customer's results. If not, please let me know the original order number and I will be happy to offer a free of charge replacement or credit.

Thank you for your email.

I see. Yes, in that case the antibody is indeed not working correctly. I apologize about this.

I wish you good luck with your research, and please let me know if I can help you with anything else.

Thank you for your reply. I'm sorry that the antibody did not work. I was wondering if the ATG8 mutant you were refering to is a deletion mutant (ATG8 protein is absent) or just point or fragmentmutation in the ATG8 protein? In case of the latter, the antibody might still be specific and just detect a band at 27 kDa ( a dimer?).

In any case, please see below for the refund information.

Your credit note ID for the refund is xxx.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.