Human Fibroblast Activation Protein alpha /FAP Antibody
Human Fibroblast Activation Protein alpha /FAP Antibody Summary
Leu26-Asp760
Accession # Q12884
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Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of Human Fibroblast Activation Protein alpha /FAP by Western Blot. Western blot shows lysates of WI-38 human lung fibroblast cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Fibroblast Activation Protein a/FAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3715) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Fibroblast Activation Protein a/FAP at approximately 97 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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Fibroblast Activation Protein alpha /FAP in Human Squamous Cell Carcinoma. Fibroblast Activation Protein a/FAP was detected in immersion fixed paraffin-embedded sections of human squamous cell carcinoma using Sheep Anti-Human Fibroblast Activation Protein a/FAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3715) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to connective tissue. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
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Fibroblast Activation Protein alpha /FAP in Human Basal Cell Carcinoma. Fibroblast Activation Protein alpha /FAP was detected in immersion fixed paraffin-embedded sections of human basal cell carcinoma using Sheep Anti-Human Fibroblast Activation Protein alpha /FAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3715) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
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Detection of Human Fibroblast Activation Protein alpha /FAP by Simple WesternTM. Simple Western lane view shows lysates of IMR‑90 human lung fibroblast cell line and WI‑38 human lung fibroblast cell line, loaded at 0.2 mg/mL. A specific band was detected for Fibroblast Activation Protein alpha /FAP at approximately 130 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human Fibroblast Activation Protein alpha /FAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3715) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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Western Blot Shows Human Fibroblast Activation Protein alpha /FAP Specificity by Using Knockout Cell Line. Western blot shows lysates of WI‑38 human lung fibroblast cell line and human FAP knockout WI‑38 human lung fibroblast cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Fibroblast Activation Protein alpha /FAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3715) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Fibroblast Activation Protein alpha /FAP at approximately 97 kDa (as indicated) in the parental WI‑38 human lung fibroblast cell line, but is not detectable in knockout WI‑38 human lung fibroblast cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Fibroblast Activation Protein alpha/FAP
FAP (also known as seprase) is a Type II transmembrane serine protease that is structurally related to dipeptidyl peptidase IV (1). FAP has substrate specificity similar to dipeptidyl peptidase IV, which is specific for N-terminal Xaa-Pro sequences, but FAP is also an endopeptidase able to degrade gelatin and Type I collagen (2). The enzymatically active form of FAP is a dimer (3). FAP has a restricted tissue distribution. It is not detectable in normal tissues or resting fibroblasts, but is highly expressed on reactive stromal fibroblasts in epithelial cancers (4), in granulation tissue during wound healing, and in bone and soft tissue sarcomas (5). Because of its expression patterns and enzymatic activities, FAP is believed to play roles in tumor invasion, tissue remodeling, and wound repair.
- Scanlan, M.J. et al. (1994) Proc. Natl. Acad. Sci. USA 91:5657.
- Park, J.E. et al. (1999) J. Biol. Chem. 274:36505.
- Pineiro-Sanchez, M.L. et al. (1997) J. Biol. Chem. 272:7595.
- Garin-Chesa, P. et al. (1990) Proc. Natl. Acad. Sci. USA 87:7235.
- Rettig, W.J. et al. (1988) Proc. Natl. Acad. Sci. USA 85:3110.
Product Datasheets
Citations for Human Fibroblast Activation Protein alpha /FAP Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
19
Citations: Showing 1 - 10
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Thy-1 plays a pathogenic role and is a potential biomarker for skin fibrosis in scleroderma
Authors: RG Marangoni, P Datta, A Paine, S Duemmel, MA Nuzzo, L Sherwood, J Varga, C Ritchlin, BD Korman
JCI Insight, 2022;0(0):.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC -
FAP is critical for ovarian cancer cell survival by sustaining NF-kappaB activation through recruitment of PRKDC in lipid rafts
Authors: B Li, Z Ding, O Calbay, Y Li, T Li, L Jin, S Huang
Cancer Gene Therapy, 2022;0(0):.
Species: Mouse
Sample Types: Whole Cell
Applications: IP -
Type IV Collagen in Human Colorectal Liver Metastases-Cellular Origin and a Circulating Biomarker
Authors: M Lindgren, G Rask, J Jonsson, A Berglund, C Lundin, P Jonsson, I Ljuslinder, H Nyström
Cancers, 2022;14(14):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Metallothionein 2A Expression in Cancer-Associated Fibroblasts and Cancer Cells Promotes Esophageal Squamous Cell Carcinoma Progression
Authors: M Shimizu, YI Koma, H Sakamoto, S Tsukamoto, Y Kitamura, S Urakami, K Tanigawa, T Kodama, N Higashino, M Nishio, M Shigeoka, Y Kakeji, H Yokozaki
Cancers, 2021;13(18):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
IL-1-driven stromal-neutrophil interactions define a subset of patients with inflammatory bowel disease that does not respond to therapies
Authors: M Friedrich, M Pohin, MA Jackson, I Korsunsky, SJ Bullers, K Rue-Albrec, Z Christofor, D Sathananth, T Thomas, R Ravindran, R Tandon, RS Peres, H Sharpe, K Wei, GFM Watts, EH Mann, A Geremia, M Attar, Oxford IBD, Roche Fibr, S McCuaig, L Thomas, E Collantes, HH Uhlig, SN Sansom, A Easton, S Raychaudhu, SP Travis, FM Powrie
Nature Medicine, 2021;0(0):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Proteomic Analyses of Fibroblast- and Serum-Derived Exosomes Identify QSOX1 as a Marker for Non-invasive Detection of Colorectal Cancer
Authors: N Ganig, F Baenke, ML Thepkayson, K Lin, VS Rao, FC Wong, H Polster, M Schneider, D Helm, M Pecqueux, AM Seifert, L Seifert, J Weitz, NN Rahbari, C Kahlert
Cancers, 2021;13(6):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Cancer-associated fibroblasts downregulate type I interferon receptor to stimulate intratumoral stromagenesis
Authors: C Cho, R Mukherjee, AR Peck, Y Sun, N McBrearty, KV Katlinski, J Gui, PK Govindaraj, E Puré, H Rui, SY Fuchs
Oncogene, 2020;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: ICC -
Extreme Diversity of the Human Vascular Mesenchymal Cell Landscape
Authors: LE Bruijn, BEWM van den Ak, CM van Rhijn, JF Hamming, JHN Lindeman
J Am Heart Assoc, 2020;9(23):e017094.
Species: Human
Sample Types: Whole Cells
Applications: ICC -
Patch repair of deep wounds by mobilized fascia
Authors: D Correa-Gal, D Jiang, S Christ, P Ramesh, H Ye, J Wannemache, S Kalgudde G, Q Yu, M Aichler, A Walch, U Mirastschi, T Volz, Y Rinkevich
Nature, 2019;0(0):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Fibroblast activation protein-positive fibroblasts promote tumor progression through secretion of CCL2 and interleukin-6 in esophageal squamous cell carcinoma
Authors: N Higashino, YI Koma, M Hosono, N Takase, M Okamoto, H Kodaira, M Nishio, M Shigeoka, Y Kakeji, H Yokozaki
Lab. Invest., 2019;0(0):.
Species: Human
Sample Types: Whole Cells
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Highly versatile cancer photoimmunotherapy using photosensitizer-conjugated avidin and biotin-conjugated targeting antibodies
Authors: N Shirasu, H Shibaguchi, H Yamada, M Kuroki, S Yasunaga
Cancer Cell Int., 2019;19(0):299.
Species: Human
Sample Types: Whole Cells
Applications: Photoimmunotherapy -
Cancer-associated fibroblasts induce antigen-specific deletion of CD8+T Cells to protect tumour cells
Authors: MA Lakins, E Ghorani, H Munir, CP Martins, JD Shields
Nat Commun, 2018;9(1):948.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry -
A synthetic urinary probe-coated nanoparticles sensitive to fibroblast activation protein ? for solid tumor diagnosis
Authors: X Feng, Q Wang, Y Liao, X Zhou, Y Wang, W Liu, G Zhang
Int J Nanomedicine, 2017;12(0):5359-5372.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Adrenergic-mediated increases in INHBA drive CAF phenotype and collagens
Authors: AS Nagaraja, RL Dood, G Armaiz-Pen, Y Kang, SY Wu, JK Allen, NB Jennings, LS Mangala, S Pradeep, Y Lyons, M Haemmerle, KM Gharpure, NC Sadaoui, C Rodriguez-, C Ivan, Y Wang, K Baggerly, P Ram, G Lopez-Bere, J Liu, SC Mok, L Cohen, SK Lutgendorf, SW Cole, AK Sood
JCI Insight, 2017;2(16):.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC -
Fibroblast activation protein augments progression and metastasis of pancreatic ductal adenocarcinoma
Authors: A Lo, CP Li, EL Buza, R Blomberg, P Govindaraj, D Avery, J Monslow, M Hsiao, E Puré
JCI Insight, 2017;2(19):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Tumor-targeting efficacy of a BF211 prodrug through hydrolysis by fibroblast activation protein-?
Authors: XP Chai, GL Sun, YF Fang, LH Hu, X Liu, XW Zhang
Acta Pharmacol. Sin., 2017;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Western Blot -
Fibroblast Activation Protein (FAP) Accelerates Collagen Degradation and Clearance from Lungs in Mice
Authors: MH Fan, Q Zhu, HH Li, HJ Ra, S Majumdar, DL Gulick, JA Jerome, DH Madsen, M Christofid, DW Speicher, WW Bachovchin, C Feghali-Bo, E Puré
J. Biol. Chem., 2016;291(15):8070-89.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Immunoprecipitation -
Antitumor effects of chimeric receptor engineered human T cells directed to tumor stroma.
Authors: Kakarla S, Chow K, Mata M, Shaffer D, Song X, Wu M, Liu H, Wang L, Rowley D, Pfizenmaier K, Gottschalk S
Mol Ther, 0;21(8):1611-20.
Species: Human
Sample Types: Whole Cells
Applications: CAR-T (Flow Cytometry) -
Suppression of antitumor immunity by stromal cells expressing fibroblast activation protein-alpha.
Authors: Kraman M, Bambrough P, Arnold J, Roberts E, Magiera L, Jones J, Gopinathan A, Tuveson D, Fearon D
Science, 0;330(6005):827-30.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry
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Human Fibroblast Activation Protein alpha
FAP Antibody ref AF3715 RetD Systems
Polyclonal Sheep . 95kd mais peut présenter
Des glycosylations d’où la taille aux alentours de
120kd!
La prochaine fois:
- Utiliser des concentrations de protéines >0,6mg/ml
- Diluer l’anticorps secondaire sheep 1/25è au lieu du
1/10è pour éviter un peu plus le bruit de fond.
- Reconstituer l’anticorps à 0,5mg/ml
Au lieu de 0,2mg/ml recommandé par R et D.
Efficacité validée dans conditions suivantes:
Dilution anticorps FAP= 1/10è (0,2mg/ml solution stock)
[CMLh]= 0,7mg/ml
Dilution anticorps 2aire sheep = 1/10è
