Af1694

Manufactured by R&D Systems

Sourced in United States, United Kingdom

AF1694 is a laboratory reagent produced by R&D Systems. It serves as a recombinant human protein, functioning as a growth factor.

Automatically generated - may contain errors

Lab products found in correlation

Af1738, by R&D Systems (2 mentions) Ab16667, by Abcam (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Ripa buffer, by Applygen (1 mentions) Diaminobenzidine solution, by ZSGB-BIO (1 mentions) Rm2235, by Leica (1 mentions) Fstl1, by R&D Systems (1 mentions) Mab1694, by R&D Systems (1 mentions)

4 protocols using af1694

FSTL1 and Ki67 Expression in Breast Cancer

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Human tumor samples from patients with breast cancer or BCBM were obtained from Cancer Hospital of Huanxing (Beijing, China). Tissues were harvested, fixed, paraffin-embedded, sectioned, and photographed. The sections were stained with hematoxylin. The expression of FSTL1 and Ki67 was determined using IHC staining. The primary antibody used was polyclonal goat anti- human FSTL1 antibody (5 µg/ml, AF1694; R&D Systems) and rabbit anti-Ki67 antibody (1:250, ab16667; Abcam).
This study was approved by Cancer Hospital of Huanxing review board, and informed consent was obtained from all patients under the protocols prescribed by Cancer Hospital of Huanxing ethics committee. All procedures performed involving human participants were in accordance with the ethical standards of the Institutional and National Research Committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

An J., Wang L., Zhao Y., Hao Q., Zhang Y., Zhang J., Yang C., Liu L., Wang W., Fang D., Lu T, & Gao Y. (2017). Effects of FSTL1 on cell proliferation in breast cancer cell line MDA-MB-231 and its brain metastatic variant MDA-MB-231-BR. Oncology Reports, 38(5), 3001-3010.

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Protein Expression Analysis in Lung and Breast Tissues

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The lung and breast tissues were lysed in RIPA buffer (Applygen, Beijing, China) containing a phosphatase inhibitor. Human FSTL1 antibody (AF1694; R&D Systems, Minneapolis, USA), Mouse FSTL1 antibody (AF1738; R&D Systems), and antibodies recognizing the following proteins were used: Phospho-p44/42 mitogen-activated protein kinase (MAPK) (#4370; Cell Signaling Technology, Boston, USA), β-actin (#4970; Cell Signaling Technology). Western blot analyses were performed following standard protocols as described previously [20 (link)].

Zhang Y., Xu X., Yang Y., Ma J., Wang L., Meng X., Chen B., Qin L., Lu T, & Gao Y. (2018). Deficiency of Follistatin-Like Protein 1 Accelerates the Growth of Breast Cancer Cells at Lung Metastatic Sites. Journal of Breast Cancer, 21(3), 267-276.

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Immunohistochemical Analysis of Breast Cancer Metastasis

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Human breast cancer samples were obtained from the Cancer Hospital of Huanxing Chaoyang District Beijing and were fixed in 4% paraformaldehyde (PFA) for 24 to 48 hours. Mouse metastatic lung tissue was dissected and fixed in PFA for 48 hours, and then stored in 70% alcohol. All samples were embedded in paraffin, and 5 µm sections were obtained using a Paraffin slicer (RM2235; Leica, Heidelberg, Germany). For hematoxylin and eosin (H&E) staining, lung tissue was stained with H&E. For immunohistochemistry, sections were deparaffinized and rehydrated, then incubated with primary antibodies (FSTL1, AF1694 and AF1738; R&D Systems) at 4℃ overnight, followed by a horseradish peroxidase secondary antibody (donkey anti-goat IgG H&L, ab97110; Abcam, Cambridge, UK) at 37℃ for 1 hour. Samples were developed using a diaminobenzidine solution (ZsBio, Beijing, China) for 2 minutes at room temperature. Sections were then dehydrated and affixed to coverslips.

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FSTL1 Quantification in Serum via ELISA

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100 μL serum samples were added to the wells of the assay plate, which was coated with 5 μg/mL human FSTL1 capture antibody (R&D system, AF1694). The plate was incubated at 4 °C overnight, then 2.5 μg/mL biotinylated human FSTL1 detection antibody (R&D system, MAB1694) was added followed by washing with wash buffer and the plate was incubated for 1 hour at room temperature. Streptavidin-HRP was added to the wells of assay plate and incubated for another 30 min at room temperature. After washing the plate again, TMB substrate was added to each well and incubated in the darkness for 15 min at room temperature. The absorbance values of the wells at 450 nm and 570 nm were obtained after the stop solution was added. The ELISA assay was performed in duplicate for every sample.

Fang Y., Zhang S., Li X., Jiang F., Ye Q, & Ning W. (2017). Follistatin like-1 aggravates silica-induced mouse lung injury. Scientific Reports, 7, 399.

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