Anti-PDI antibody - ER Marker (ab31811)

Dear xxxx,  

I think that ab31811 (rabbit polyclonal) antibody agains PDI is more suitable for us because they was already tested with Human brain and they are polyclonal (it is critical for our goal) >You wrote Alternatively, as discussed yesterday, whilst ab31811  has not been as well characterized as ab2792 (what does it mean, "not been as well characterized"?), it has been used to perform Western blotting with Human brain (which kind of brain area? cortex, putamen, cerebellum?) whole tissue lysate in house and I can obtain a detailed protocol of how this is performed if you are still interested in this antibody. It will be great to have a full protocol from this experiment. And as I understood if I have any problems obtaining the results with this antibody with human brain samples, I am entitled to an alternative antibody or a refund. Is that correct?  It is very important to discuss this point with my supervise.

Best regards,

ANSWER:

Sorry for the dely in getting back to you. I now have further information concerning Anti-PDI antibody (ab31811).

The human brain tissue lysate used for the Western blot presented on the datasheet of ab31811 was taken from the parietal lobe.

The protocol used to perform this Western blot is as follows: 1. Tissue lysate prepared 2. Lysate (20-40 µg) loaded on SD-page gel and transferred to Nitrocellulose membrane  3. Membrane blocked using 5% BSA in 0.1% TBST, for 1 hour at room temperature 4. Primary diluted to 1 µg/ml in 5% BSA TBST, incubated with the membrane overnight at 4°C with gentle rocking 5. Was step performed: 0.1% TBST 2 brief rinses followed by 1x15 min then 3x10 min at room temperature. 6. Secondary antibody incubated with membrane: IR Dye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution in 5% BSA 0.1% TBST, at room temperature for 1 hour, with gentle rocking. 7. Repetition of wash step 5

I hope this information has been of help. If you need any further information or help in ordering please do not hesitate to contact me again.

Dear xxxxx,  

I think that ab31811 (rabbit polyclonal) antibody agains PDI is more suitable for us because they was already tested with Human brain and they are polyclonal (it is critical for our goal) >You wrote Alternatively, as discussed yesterday, whilst ab31811  has not been as well characterized as ab2792 (what does it mean, "not been as well characterized"?), it has been used to perform Western blotting with Human brain (which kind of brain area? cortex, putamen, cerebellum?) whole tissue lysate in house and I can obtain a detailed protocol of how this is performed if you are still interested in this antibody. It will be great to have a full protocol from this experiment. And as I understood if I have any problems obtaining the results with this antibody with human brain samples, I am entitled to an alternative antibody or a refund. Is that correct?  It is very important to discuss this point with my supervise. Best regards,

ANSWER:

Thank you for contacting us again.

When I said that ab31811 had not been charaterised as well as ab2792 I simply meant that this antibody had not been used and referenced in as many publications and had not had as many reviews by our other customers. This does not mean that it will perform any differently. I think ab31811 would be suitable for the experiment you will be performing.

I am currently investigating exactly which type of brain sample was used with ab31811 to obtain the blot presented on the datasheet and as soon as I have the answer I will get back to you. You are correct in thinking that if you have problems obtaining the results expected with Western blotting of human samples you would be entitled to a refund.

We usually deal with such cases (which are quite rare) by initially looking into the experiment you have been performing by getting you to fill out a brief questionnaire (5-10 min max) in order to understand the problem and investigate the case internally if need be, then if we can't find anything to help you otimise the protocol we would offer a free of charge replacement or a refund, whichever you prefer. We are very flexible and understand that you know what you are doing so therefore usually the questionnaire is just to establish if we need to initiate some additional testing on our side. We do not try to withhold refunds.

I hope this information has been of help, if you need any further information please do not hesitate to ask. I will let you know once I have received confirmation of the obtain sample used and the protocol used.

Phone call asking if any publications were available for Anti-PDI antibody ab31811. They wish to use it for Western blotting with human brain samples. Was also interested in Calnexin antibodies for the same application and sample type.

ANSWER:

Thank you for contacting us. I have looked into the products that we have available for the species and application you are interested in (Western blotting with human brain samples) for PDI and Connexin.

1. PDI

There are two antibodies which look like they would be promising for your research: ab2792 is a mouse monoclonal which has been used in the following publications to perform Western blotting and immunoprecipitation, although not with the same tissue samples you are hoping to analyse:

Western blotting with human granulocytes, antibody used as loading control:

Petheo GL et al. Molecular and functional characterization of Hv1 proton channel in human granulocytes. PLoS One 5:e14081 (2010). WB; Human. PubMed: 21124855

Immunoprecipitaiton illustrating the association of PDI with TAP in M553 (melanoma cell line):

Rizvi SM & Raghavan M Mechanisms of Function of Tapasin, a Critical Major Histocompatibility Complex Class I Assembly Factor. Traffic 3, 332-347, (2009). IP; Human. PubMed: PubMed: 20070606

This antibody has also been used with several human cell lines to perform Western blotting with quite clean results as well as one report of using Human Tissue lysate - whole (Brain):

http://www.abcam.com/index.html?datasheet=2792&tab=abreviews&intabreviewid=13102

Alternatively, as discussed yesterday, whilst ab31811 (rabbit polyclonal) has not been as well charaterised as ab2792, it has been used to perform Western blotting with Human brain whole tissue lysate in house and I can obtain a detailed protocol of how this is performed if you are still interested in this antibody.

Both of these antibodies have been tested for human samples and Western blotting and as such would be covered by our Abpromise guarantee, more information of which can be found from the link below. This means that if you have any problems obtaining the results you would expect with this antibody with your human brain samples, and we are unable to help you obtain better results, you would be entitled to an alternative antibody or a refund.

www.abcam.com/abpromise

2. Connexin

As there are so many different forms of connexin, in order to suggest the best antibody for you could you please specify which form you are wanting to study? We have antibodies against:

Connexin 36/GJA9, 30.3/GJB4, 40.1/GJD4, 59/GJA10, 26/GJB2, 30/GJB6, 62GJA10, 46/GJA3, 31/GJB3, 50/GJA8, 23/GJE1, 31.1/GJB5, 47GJC2, 45/GJC1

I hope this information has been of help. If you'd like more help in selecting a suitable anti-connexin antobody, or would like any other information please do let me know.