Anti-Fibroblast activation protein, alpha 抗体

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody (AB53066)

Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue sections labeling Fibroblast activation protein, alpha with ab53066 (without peptide-right and with peptide- left).

• WB

Unknown

Western blot - Anti-Fibroblast activation protein, alpha antibody (AB53066)

Blocking buffer : 5% (w/v) non-fat dry milk in TBST.

Primary antibody dilution buffer : 5%（w/v）non-fat dried milk，0.1%（v/v）, Tween-20 in TBST.

Secondary antibody dilution buffer : 5%（w/v）non-fat dried milk，0.1%（v/v），Tween-20 in TBST.

12% SDS gel. Nitrocellulose membrane.

Blocking : Room temperature for 2 hours or overnight at 4°C. Then wash 3x for 5 minutes with 0.05% blocking buffer.

Primary antibody incubation : diluted in TBST at 1/500. Incubate overnight with 4 degrees shaking. Then, in 0.05% TBST, wash membrane 3-4 times for 10min.

Secondary antibody incubation : diluted in TBST at 1/2000. Incubate 37°C for 1 hour. Then, in 0.05% TBST, wash membrane 3-4 times for 10min.

ECL development.

All lanes:

Western blot - Anti-Fibroblast activation protein, alpha antibody (ab53066) at 1/500 dilution

Lane 1:

LOVO cell extract

Lane 2:

LOVO cell extract at 40 µg with immunising peptide

Predicted band size: 88 kDa

Observed band size: 90 kDa

false

• IHC-P

CiteAb

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody (AB53066)

-Fibroblast activation protein immunohistochemistry using Anti-Fibroblast activation protein, alpha antibody ab53066. Publication image and figure legend from Wu, J., Feng, Z., et al., 2022, Nat Commun, Pubmed 35115492.

The ferroptosis inducer IKE decreased fibroblast populations in RA synovium.a Representative immunohistochemical staining of FAPα in hyperplastic rheumatoid synovium of 26 RA and 21 OA patients. Scale bars, 50 μm. b Representative fluorescent multiplex IHC staining and scoring of joints labeled with anti-F4/80 (red), anti-FAPα (green), and DAPI (blue). Scale bars, 100 μm. n = 413 (NC), 3698 (CIA), 3469 (CIA + IKE) cells from 3 independent joints for each group. ****p < < 0.0001; ns, P = 0.6849; one-way ANOVA followed by multiple comparisons. c Cell death in FAPα+ fibroblasts, CD68+ macrophages, CD31+ endothelial cells, CD3+ T cells, or CD19+ B cells isolated from hyperplastic synovium treated with 0.125 μm RSL3 for 6 h, quantified by SYTOX staining followed by flow cytometry. Cell death (d) and lipid ROS production (e) in circulating fibrocytes from PBMCs and synovial fibroblasts from inflamed joint fluid of RA patients treated with 0.125 μm RSL3 for 18 h (cell death) or 6 h (lipid ROS). ****p < < 0.0001, ***P = 0.0001; two-tailed t-test. n = 6 patients. f Representative fluorescent multiplex IHC staining of retained hyperplasia synovium of CIA model mice treated with or without IKE and labeled with anti-F4/80 antibody (red), anti-FAPα antibody (green), and DAPI (blue). Scale bars, 200 μm. g The number of F4/80+ macrophages within the 50 μm radium of FAPα+ fibroblasts in joints of CIA mice. *p < < 0.05; ns, P = 0.00878; two-tailed t-test. n = 3 joints. Data are presented as mean ± SD. Source data are provided as a Source data file.

• IHC-P

CiteAb

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody (AB53066)

-Fibroblast activation protein immunohistochemistry using Anti-Fibroblast activation protein, alpha antibody ab53066. Publication image and figure legend from Errarte, P., Guarch, R., et al., 2016, PLoS One, Pubmed 28033421.

FAP immunohistochemical expression in primary and metastatic CCRCC.Clear cell renal cell carcinoma arranged in nests (A) show FAP-positive cancer-associated fibroblasts delineating tumor lobes (B). Lymph node metastatic tumor growing more diffusely without recognizable architecture (C) displays FAP-positive cancer-associated fibroblasts intimately intermingled with tumor cells (D).

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-Fibroblast activation protein, alpha antibody (AB53066)

-Fibroblast activation protein Immunocytochemistry-immunofluorescence using Anti-Fibroblast activation protein, alpha antibody ab53066. Publication image and figure legend from Kang, M. K., Jiang, F., et al., 2023, Cancers (Basel), Pubmed 37444482.

CTHRC1 activated pancreatic stellate cells (PSCs) in vitro. Activation of PSC after treatment with PBS, CTHRC1 (1.7 nM), or TGF-β (2.1 nM) for 24 h demonstrated by the protein expression of FAP and α-SMA in PSCs using (A) Western blot and (B) immunofluorescence (IF) assay, by (C) quantification of lipid droplets in PSC cytoplasm using Oil Red O staining (decreased optical density (O.D.) of lipid droplets indicates PSC activation), and by (D) metastatic capacity of PSCs using migration/invasion assays. To confirm the activating effects of CTHRC1 on PSCs, PSCs were treated with PBS, CTHRC1 (1.7 nM), and/or α-CTHRC1 (anti-CTHRC1 mAb, 69.4 nM) for 24 h, (E) concentrations of collagen I and fibronectin were measured using indirect ELISA, and (F) metastatic capacity of PSCs was determined using migration/invasion assays. p-values : *, **, *** and **** indicate p < < 0.05, 0.01, 0.001 and 0.0001, respectively.

• WB

CiteAb

Western blot - Anti-Fibroblast activation protein, alpha antibody (AB53066)

Fibroblast activation protein, alpha western blot using anti-Fibroblast activation protein, alpha antibody ab53066. Publication image and figure legend from Evrard, S. M., Lecce, L., et al., 2016, Nat Commun, PubMed 27340017.

ab53066 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab53066 please see the product overview.

Oxidative stress promotes EndMT and has additive effects to TGF-β.(a) Representative dihydroethidium (DHE) microscopy images of thoracic aortic sections from tamoxifen-induced end.SclCreERT;R26RstopYfp;ApoE-/- mice after 8 versus 18 weeks HFD demonstrating in situ superoxide production. Analysis by one-way ANOVA; overall p<0.0001. (b) TUNEL assay on atherosclerotic lesions from tamoxifen-induced end.SclCreERT;R26RstopYfp;ApoE-/- mice after 8 versus 18 weeks HFD demonstrating apoptotic Yfp+ cells. For a and b : L=lumen; scale bars, 100 μm. Insets at higher magnification as indicated. Arrows indicate co-positive cells. Staining performed on at least three spatially separated thoracic aortic sections per mouse (n=5 mice per group). Control experiments from non-atherosclerotic mice shown in Supplementary Fig. 8. (c) Phase contrast microscopy of HUVECs grown in basal media (ctrl) or following exposure to TGF-β and/or H2O2 for 5 days. Scale bars, 100 μm. (d) Heatmap showing expression of endothelial genes (top), non-endothelial genes also downregulated in EndMT/EMT, genes known to be upregulated in EndMT/EMT and mesenchymal genes (bottom) for HUVECs exposed to control conditions (basal media—unstimulated), TGF-β or TGF-β plus 200 μM H2O2. Because there are no gene sets for 'EndMT' or 'fibroblast' in databases like GO, KEGG, Reactome or MSigDB, this gene list was compiled by extensive literature search (Supplementary Table 2). (e) FAP protein expression by western blot with quantification in HUVECs exposed to control (ctrl) conditions (unstimulated) or TGF-β, with and without H2O2 at 100 or 200 μM. FAP ladder (L) represents 100 kDa (upper) and 75 kDa (lower intense band). FAP expected molecular weight/size is 90 kDa. GAPDH ladder represents 37 kDa. GAPDH expected molecular weight/size is 37 kDa. (f) FAP RNA levels assessed by qRT-PCR under identical conditions. (g) CD31 protein by western blot with quantification. CD31 ladder (L) represents 150 kDa (upper) and 100 kDa (lower intense band). CD31 expected molecular weight/size is 130 kDa. GAPDH ladder represents 37 kDa. GAPDH expected molecular weight/size is 37 kDa. (h) CD31 RNA levels assessed by qRT-PCR is presented for HUVECs under identical conditions. Data in e–h analysed by two-way ANOVA with complete results in Supplementary Table 3. (i) Cell migration for unstimulated HUVECs versus after treatment with TGF-β only or TGF-β+200 μM H2O2 for 5 days. Analysis by one-way ANOVA; overall P=0.015. (j) Cell invasion for unstimulated HUVECs versus after treatment with TGF-β only or TGF-β+200 μM H2O2 for 5 days. Analysis by one-way ANOVA; overall p<0.0001. n=6–9 for qRT-PCR and 3–6 for western blots. Migration and invasion determined from three independent experiments with each experiment having n=3. NS, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

false

• WB

CiteAb

Western blot - Anti-Fibroblast activation protein, alpha antibody (AB53066)

-Fibroblast activation protein western blotting using Anti-Fibroblast activation protein, alpha antibody ab53066. Publication image and figure legend from Zhang, S., Wang, J., et al., 2024, Cell Biosci, Pubmed 39605072.

CAFs exhibit increased glycolytic activity and lactate production. A Extract bright field images of primary CAFs and PFs from three pairs of patients. Scale bars, 200 μm. B Extracting bright field diagrams of TGF-β-treated (10ng/ml) NIH-3T3. Scale bars, 10 μm. C Western blot results of CAFs-related markers were used to prove that the obtained cells were CAFs. β-actin was used as a control. D, E Clear flow chart of CAF and PF, as well as NIH-3T3 and its induced cells culture medium after the same cultivation time, as well as statistical chart of the pH value of the culture medium. Student t-test. F ECAR experiment was used to detect the release rate of acidic metabolites produced by cells. G Glucose consumption experiment was used to detect the rate of glucose consumption and uptake by cells. H Detecting the lactate secretion levels of CAFs, PFs, NIH-3T3 and their induced cells separately. I, J RT-qPCR and western blot results of glycolysis-related markers. β-actin and GAPDH were used as a control. Independent experiments (in vitro) were performed in triplicate. Data are presented as mean ± SD. *p < < 0.05, **p < < 0.01, ***p < < 0.001, ****p < < 0.0001

false

• WB

CiteAb

Western blot - Anti-Fibroblast activation protein, alpha antibody (AB53066)

-Fibroblast activation protein western blotting using Anti-Fibroblast activation protein, alpha antibody ab53066. Publication image and figure legend from Kang, M. K., Jiang, F., et al., 2023, Cancers (Basel), Pubmed 37444482.

CTHRC1 activated pancreatic stellate cells (PSCs) in vitro. Activation of PSC after treatment with PBS, CTHRC1 (1.7 nM), or TGF-β (2.1 nM) for 24 h demonstrated by the protein expression of FAP and α-SMA in PSCs using (A) Western blot and (B) immunofluorescence (IF) assay, by (C) quantification of lipid droplets in PSC cytoplasm using Oil Red O staining (decreased optical density (O.D.) of lipid droplets indicates PSC activation), and by (D) metastatic capacity of PSCs using migration/invasion assays. To confirm the activating effects of CTHRC1 on PSCs, PSCs were treated with PBS, CTHRC1 (1.7 nM), and/or α-CTHRC1 (anti-CTHRC1 mAb, 69.4 nM) for 24 h, (E) concentrations of collagen I and fibronectin were measured using indirect ELISA, and (F) metastatic capacity of PSCs was determined using migration/invasion assays. p-values : *, **, *** and **** indicate p < < 0.05, 0.01, 0.001 and 0.0001, respectively.

false