Anti-Fibroblast activation protein, alpha antibody - Catalytic insert (ab28246)

Hi, I want to buy Fibroblast activation protein antibody, and I saw there are 4 products on your website. Ab28243 Catalytic domain Ab28246 Catalytic insert Ab28245 Spacer Ab28244 Stalk I want to do immunofluorescence in rat fibroblast to see if my treatment can activate fibroblast or not. I want to use FAP as a marker for the activated fibroblast. So which antibody do you suggest? Thanks!

ANSWER:

Thank you for contacting Abcam.

I would recommend using ab28244 or ab28246 as each of these is guaranteed to work in IF on rat tissues under our Abpromise.

The immunogen for ab28244 Fibroblast activation protein, alpha antibody starts at the beginning of the stalk region.This region starts at amino acid 30 of the 760 amino acid sequence of human FAP-alpha, and follows the transmembrane domain. Cleavage of the stalk region releases the shed form of FAP-alpha.

ab28246(Fibroblast activation protein, alpha antibody - Catalytic insert) will recognize the 759 amino acid sequence of human FAP-alpha which has an insert in the catalytic domain.

These products are covered by our Abpromise guarantee. If you experience any issues with these products we offer scientific support, replacement or refund under our Abpromise guarantee, for a period of 6 months from purchase, if used according to specifications listed on our datasheet.

Phone call: the customer does not see a signal in Western blot on human primary fibroblasts and would like to know more information about the sample preparation

ANSWER:

Merci pour votre appel de hier.

Je reçu une réponse du laboratoire. Effectivement, le tampon rigoureux d'inhibition de protéases dans le tampon de lyse est seulement requis pour maintenir le FAB-a dans une forme latente. La raison la plus probable de ne pas voir des bandes dans le Western blot est que les cellules ne produisent pas de la FAP-a (pas toutes ces cellules produisent cette protéine). Le mieux serait donc d'utiliser un contrôle positive pour en être sure. Il existent des protéines recombinantes FAP-a (ab79623) ou alternativement, le laboratoire recommande comme contrôle positive des cellules de mélanome humain (ab29339, ab14634, ab14638), qui eux produisent des quantités raisonnables de FAP-a.

Click here (or use the following: http://www.abcam.com/index.html?datasheet=79623).

Click here (or use the following: http://www.abcam.com/index.html?datasheet=29339).

Click here (or use the following: http://www.abcam.com/index.html?datasheet=14634).

Click here (or use the following: http://www.abcam.com/index.html?datasheet=14638).

Veuillez me dire ce que vous en pensez. Etes vous vraiment sûre de l'expression dans vos cellules?

Je me réjouis d'avoir de vos nouvelles et vous souhaite en attendant un très bon weekend.

Dear technical support: The customer have inquires about ab28245 and ab28244. He would like to know what difference of immunogene between them. What is difference between spacer regions and talk region? He would like to perform in flowcytometry. Thank you for your assistance.

ANSWER:

Thank you for your enquiry.

The following information is from the FAQ (frequently asked questions) tab from the datasheet of ab28244:

The descriptive terms for these antibodies are based on comparison with the dipeptidyl peptidase family, especially DPP-4. FAP-alpha starts with a short cytoplasmic domain followed by a transmembrane domain.

ab28244 is rasied against the stalk region. This region starts at amino acid 30 of the 760 amino acid sequence of human FAP-alpha, and follows the transmembrane domain. Cleavage of the stalk region releases the shed form of FAP-alpha. The immunogen for ab28244 Fibroblast activation protein, alpha antibody starts at the beginning of the stalk region.

Between the stalk region and the catalytic domain is the spacer region. Antibody ab28245 (Fibroblast activation protein, alpha antibody - Spacer region) detects in the middle of this spacer region, it starts shortly after amino acid 250.

We also have another antibody, ab28243 (Fibroblast activation protein, alpha antibody - Catalytic domain) which detects in the catalytic domain and near the carboxyterminal end of the whole protein. The 759 amino acid sequence of human FAP-alpha has an insert in the catalytic domain, and ab28246 (Fibroblast activation protein, alpha antibody - Catalytic insert) will recognize this form specifically.

I hope this information will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

Hellow! I want to ask you a question about this antibody,'stalk region','spacer region'and 'catalytic insert',what do they mean?I want to know which section they represent in FAPa protein,i am looking forword to your answers.

ANSWER:

Thank you for your enquiry.

The descriptive terms for these antibodies are based on comparison with the dipeptidyl peptidase family, especially DPP-4. FAP-alpha starts with a short cytoplasmic domain followed by a transmembrane domain.

The stalk region, which starts at amino acid 30 of the 760 amino acid sequence of human FAP-alpha, follows the transmembrane domain. Cleavage of the stalk region releases the shed form of FAP-alpha. RP1-FAP-alpha (ab28244 Fibroblast activation protein, alpha antibody - Stalk region) starts at the beginning of the stalk region.

Between the stalk region and the catalytic domain is the spacer region. RP2-FAP-alpha ab28245 (Fibroblast activation protein, alpha antibody - Spacer region) is in the middle of this spacer region, it starts shortly after amino acid 250.

RP3-FAP-alpha (ab28243 Fibroblast activation protein, alpha antibody - Catalytic domain) is in the catalytic domain and near the carboxyterminal end of the whole protein. The 759 amino acid sequence of human FAP-alpha has an insert in the catalytic domain, and ab28246 (Fibroblast activation protein, alpha antibody - Catalytic insert) will recognize this form specifically.

I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

You have the following antibodies for this protein and each of their immunogen sequences is to a different portion of the protein.

Ab28243 Catalytic domain

Ab28246 Catalytic insert

Ab28245 Spacer

Ab28244 Stalk

I want to find an antibody that will work well in a western blot and in rat. I would like to know where on the protein these immunogen sequences lie. Not necessarily the specific amino acid sequence. For example: where is the stalk region and the spacer region?

Also, would you recommend any of these antibodies over the other one for my purposes? I didn't find that any of them had western blot data on their data sheets.

ANSWER:

Thank you for your enquiry. The descriptive terms for these antibodies are based on comparison with the dipeptidyl peptidase family, especially DPP-4. FAP-alpha starts with a short cytoplasmic domain followed by a transmembrane domain. The stalk region, which starts at amino acid 30 of the 760 amino acid sequence of human FAPa, follows the transmembrane domain. Cleavage of the stalk region releases the shed form of FAPa. RP1-FAPa starts at the beginning of the stalk region. Between the stalk region and the catalytic domain is the spacer region. RP2-FAPa is in the middle of this spacer region, it starts shortly after amino acid 250. RP3-FAPa is in the catalytic domain and near the carboxyterminal end of the whole protein. The 759 amino acid sequence of human FAPa has an insert in the catalytic domain, and RP4-FAPa will recognize this form specifically.

All four antibodies will detect rat FAPa in Western blots. RP2-FAPa and RP3-FAPa are probably most suitable for this researcher's needs. Should you have any further questions then please do not hesitate to get back in touch with me.