Overview

  • Product name
    Anti-Fibroblast activation protein, alpha antibody
    See all Fibroblast activation protein, alpha primary antibodies
  • Description
    Rabbit polyclonal to Fibroblast activation protein, alpha
  • Host species
    Rabbit
  • Specificity
    This antibody recognizes the Stalk Region of both long isoforms of Fibroblast activation protein, alpha but does not recognize other DPP family members.
  • Tested applications
    Suitable for: ICC/IF, IHC-P, Flow Cyt, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Fibroblast activation protein, alpha. The exact sequence is proprietary.

Properties

Applications

Our Abpromise guarantee covers the use of ab28244 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 1 - 10 µg/ml.
IHC-P Use at an assay dependent concentration. PubMed: 18403594
Flow Cyt 1/50. PubMed: 20361950

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/1000 - 1/5000. Detects a band of approximately 110-120 kDa (predicted molecular weight: 88 kDa). 1/1000 when using colorimetric substrates such as BCIP/NBT - 1/5000 when using chemiluminescent substrates. Note: EDTA/EGTA treatment of tissues or lysates is required to see latent zymogen. Dilution optimised using Chromogenic detection.

Target

  • Function
    In association with DPP4 is involved in the pericellular proteolysis of the extracellular matrix (ECM), the migration and invasion of endothelial cells into the ECM. May have a role in tissue remodeling during development and wound healing, and may contribute to invasiveness in malignant cancers.
  • Tissue specificity
    Fibroblast specific.
  • Sequence similarities
    Belongs to the peptidase S9B family.
  • Post-translational
    modifications
    N-glycosylated.
    The N-terminus may be blocked.
  • Cellular localization
    Cell membrane. Cell projection > lamellipodium membrane. Cell projection > invadopodium membrane. Found in cell surface lamellipodia, invadopodia and on shed vesicles. Colocalized with DPP4 in invadopodia and lamellipodia of migratory activated endothelial cells in collagenous matrix. Colocalized with DPP4 on endothelial cells of capillary-like microvessels but not large vessels within invasive breast ductal carcinoma.
  • Information by UniProt
  • Database links
  • Alternative names
    • 170 kDa melanoma membrane bound gelatinase antibody
    • 170 kDa melanoma membrane-bound gelatinase antibody
    • DPPIV antibody
    • FAP antibody
    • FAPA antibody
    • Fibroblast activation protein alpha antibody
    • Integral membrane serine protease antibody
    • SEPR_HUMAN antibody
    • Seprase antibody
    see all

Images

  • All lanes : Anti-Fibroblast activation protein, alpha antibody (ab28244) at 1/1000 dilution

    Lane 1 : Recombinant Human FAPalpha at 0.08 µg
    Lane 2 : Recombinant Human FAPalpha at 0.04 µg
    Lane 3 : Recombinant Human FAPalpha at 0.02 µg
    Lane 4 : Recombinant Human FAPalpha at 0.01 µg
    Lane 5 : Recombinant Human FAPalpha at 0.005 µg

    Predicted band size: 88 kDa

  • ICC/IF image of ab28244 stained MEF1 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28244, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Boulding T  et al. LSD1 activation promotes inducible EMT programs and modulates the tumour microenvironment in breast cancer. Sci Rep 8:73 (2018). Read more (PubMed: 29311580) »
  • Yang Z  et al. Dicer reprograms stromal fibroblasts to a pro-inflammatory and tumor-promoting phenotype in ovarian cancer. Cancer Lett 415:20-29 (2018). WB, IF . Read more (PubMed: 29199004) »
See all 18 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Mouse breast cancer)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris ph9 / citrate pH6
Permeabilization
Yes - Tween 20 (0.1%)
Specification
Mouse breast cancer
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 10 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse breast tumors)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
50 µg
Specification
Mouse breast tumors
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Aug 10 2018

Answer

Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As we discussed on the phone, I would like to collect a few more details from you in regards to the samples and protocol used with the antibody. To this end, I have attached aquestionnaire. This should only take 5-10 minutes to complete and if you could include any images of the staining that would be very helpful.

OnceI havereceive the completed questionnaire,I will look at the protocol and see if there are any suggestionsI can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary.

If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

Read More

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.

Read More

Answer

Thank you for contacting Abcam.
We do not know of any publications which have used this product in IHC-P in mouse tissues.
A good starting point for determining the optimal concentration is to use the dilution used in the earlier referenced paper, 1:600. Of course you will need to titer out the best concentration but using this reference for your experiments will give you an excellent start.
I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Read More

Answer

Thank you for contacting us.

This product was used in IHC-P in the following publication: Bhati R et al. Molecular characterization of human breast tumor vascular cells. Am J Pathol 172:1381-90 (2008). IHC-P; Human. (PubMed: 18403594) (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2329846/?tool=pubmed)
Where it is referenced as Rabbit polyclonal antibody to fibroblast activation protein (FAP), α-Stalk region (Abcam, Cambridge, MA) and was used at 1:600 dilution.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your enquiry and your interest in our products.

The tested applications and the species are normally listed on the on-line product datasheets. Generally, we test our antibodies in human, mouse and rat samples - very few antibodies are characterized in specifically in porcine. However, it does not necessary mean that they are unsuitable in this species but we have no preliminary data yet to support the cross-reactivity.

I have conducted a search for you and found some antibodies (ab28244, ab44971, ab51824) which recognize fibroblasts and are suitable for Flow cytometry but no cross-reactivity data with porcine so far. We can certainly offer a discount code for testings and the Terms and Conditions of this offer can be found at the following site: https://www.abcam.com/collaborationdiscount.

If you are interested in any particular fibroblast markers, you may wish to conduct a search using our website and check some publications using PubMed websites.

I hope this helps and if I can assist further, please do not hesitate to contact me.

Read More

Answer

Thank you for contacting Abcam regarding ab28244. Unfortunately the immunogen information for ab28244 is considered proprietary and I do not have access to that information. Please accept my apologies for not being able to provide you with the information you required. If there is anything else I can do help you, please do not hesitate to contact me.

Read More

Answer

Thank you for your reply. I can confirm that only ab28244 (Anti-Fibroblast activation protein, alpha antibody - Stalk region) is tested and guaranteed for flow cytometry and I therefore recommend this antibody for your experiments. Please find a publication that used this antibody in flow cytometry on the datasheet: datasheet=28244.https://www.abcam.com/index.html?datasheet=28244 https://www.abcam.com/index.html?datasheet=28244. 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 and is extracellular. 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. This antibody recognizes the Stalk Region of both long isoforms of Fibroblast activation protein-alpha. 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. I hope this will be helpful to you. Please fell for to contact me again in case you have further questions.

Read More

Answer

Thank you for contacting us. I can confirm that ab28244 is tested and guaranteed for FACS and IHC-P in mouse, human and rat by our Abpromise. and therefore should be suitable to isolate cells that express FAP. This antibody recognizes the Stalk Region of both long isoforms of Fibroblast activation protein-alpha. The stalk region, which starts at amino acid 30 of the 760 amino acid sequence of human FAP-alpha, follows the transmembrane domain and is extracellular. Cleavage of the stalk region releases the shed form of FAP-alpha. We have no data concerning the recognition of the cleaved FAP by ab28244. We are not specialized in the field of pancreatic stellate cells and do not know if this antibody is suitable to isolate these cells specifically. We do not want to recommend a product that may be unsuitable. We encourage customers to consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about their specific research interests. I am sorry that I could not be more helpful, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

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1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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