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Anti mif

Manufactured by Cell Signaling Technology
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Anti-MIF is a laboratory reagent produced by Cell Signaling Technology. It is an antibody designed to detect the presence and quantity of the protein macrophage migration inhibitory factor (MIF) in biological samples.

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Lab products found in correlation

Anti rabbit hrp, by Thermo Fisher Scientific (2 mentions) Precast gel, by Thermo Fisher Scientific (2 mentions) Anti beta actin, by Fortis Life Sciences (2 mentions) Ibright imager, by Thermo Fisher Scientific (2 mentions) Ripa buffer, by Boston BioProducts (2 mentions) Protease and phosphatase inhibitor, by Roche (2 mentions) Hrp conjugated goat anti rabbit and anti mouse antibodies, by Cell Signaling Technology (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Anti erk, by Cell Signaling Technology (1 mentions) Fura 2 am, by Thermo Fisher Scientific (1 mentions) Psuper vector, by Oligoengine (1 mentions) Anti pc1 7e12, by Santa Cruz Biotechnology (1 mentions) Anti mtor, by Cell Signaling Technology (1 mentions) Anti p mtor, by Cell Signaling Technology (1 mentions) Anti p erk, by Cell Signaling Technology (1 mentions) Ionomycin, by Merck Group (1 mentions) Lympholyte h, by Cedarlane (1 mentions) Anti f actin, by Santa Cruz Biotechnology (1 mentions) Confocal laser scanning microscope, by Leica Microsystems (1 mentions)

4 protocols using anti mif

1

Western Blot Analysis of MIF Protein

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Protein lysates were prepared with ice-cold RIPA buffer (Boston Bio Products) supplemented with protease and phosphatase inhibitors (Roche). Protein lysates were separated using SDS-polyacrylamide gel electrophoresis (pre-cast gels, Thermo Fisher) and transferred to a nitrocellulose membrane. After blocking the membrane with 5% milk in PBST (PBS with 0.1% Tween20, Sigma), the membranes were incubated overnight at 4°C with anti-MIF (Cell Signaling) and anti-Beta Actin (Bethyl) antibodies, diluted in PBST with 5% BSA. The blots were washed 3 times and then incubated with secondary antibodies (anti-Rabbit HRP, Thermo Fisher), diluted in PBST with 5% milk. Finally, the membranes were incubated with ECL substrate (Pierce) for 1 min and exposed for signal detection with the iBright Imager (Thermo Fisher).
Tessaro F.H., Ko E.Y., De Simone M., Piras R., Broz M.T., Goodridge H.S., Balzer B., Shiao S.L, & Guarnerio J. (2022). Single-cell RNA-seq of a soft-tissue sarcoma model reveals the critical role of tumor-expressed MIF in shaping macrophage heterogeneity. Cell reports, 39(12), 110977.
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2

Western Blot Analysis of MIF Protein

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Protein lysates were prepared with ice-cold RIPA buffer (Boston Bio Products) supplemented with protease and phosphatase inhibitors (Roche). Protein lysates were separated using SDS-polyacrylamide gel electrophoresis (pre-cast gels, Thermo Fisher) and transferred to a nitrocellulose membrane. After blocking the membrane with 5% milk in PBST (PBS with 0.1% Tween20, Sigma), the membranes were incubated overnight at 4°C with anti-MIF (Cell Signaling) and anti-Beta Actin (Bethyl) antibodies, diluted in PBST with 5% BSA. The blots were washed 3 times and then incubated with secondary antibodies (anti-Rabbit HRP, Thermo Fisher), diluted in PBST with 5% milk. Finally, the membranes were incubated with ECL substrate (Pierce) for 1 min and exposed for signal detection with the iBright Imager (Thermo Fisher).
Tessaro F.H., Ko E.Y., De Simone M., Piras R., Broz M.T., Goodridge H.S., Balzer B., Shiao S.L, & Guarnerio J. (2022). Single-cell RNA-seq of a soft-tissue sarcoma model reveals the critical role of tumor-expressed MIF in shaping macrophage heterogeneity. Cell reports, 39(12), 110977.
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3

Plasmid-mediated Silencing of PKD2

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Culture media, saline buffers and plastic material were obtained from EuroClone (Milan, Italy). Anti-P-mTOR, anti-mTOR, anti-P-ERK, anti-ERK, and anti-MIF antibodies were purchased from Cell Signaling Technology (EuroClone). Anti PC1 7E12, anti-β-actin and anti-NFkB antibodies were bought from Santa Cruz Biotechnology (Milan, Italy). Anti-CD3 (OKT3) monoclonal antibody was purchased from Invitrogen (Thermo Fisher Scientific, Milan, Italy). The anti-PC2 N-ter antibody was kindly provided by Prof. Stefan Somlo (Yale University, CT, USA). Enhanced chemiluminescent substrates for Western blotting, HRP-conjugated goat anti-rabbit and anti-mouse antibodies were purchased from Cell Signaling Technology (EuroClone). Lympholyte®-H and phytohaemagglutinin (PHA) were obtained from Cedarlane Laboratories (Tebu-Bio, Milan, Italy) and EuroClone, respectively. Interleukin-2 (IL-2) and Fura 2-AM were purchased from Invitrogen. Platelet-activating factor (PAF), ATP and ionomycin were purchased from Sigma-Aldrich (Milan, Italy). The recombinant plasmid for PKD2 silencing (TRPP2-siRNA) was constructed in our laboratory by using the pSuper vector (Oligoengine, Seattle, WA) as previously described [12 (link)].
Magistroni R., Mangolini A., Guzzo S., Testa F., Rapanà M.R., Mignani R., Russo G., di Virgilio F, & Aguiari G. (2019). TRPP2 dysfunction decreases ATP-evoked calcium, induces cell aggregation and stimulates proliferation in T lymphocytes. BMC Nephrology, 20, 355.
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4

Immunofluorescence Analysis of Cellular Morphology

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To confirm morphological changes, cells were washed thrice with PBS, fixed in 4% paraformaldehyde (PFA) for 10 min at room temperature, and permeabilized with PBS containing 0.25% Triton X-100 (PBST) for 10–15 min at room temperature. After three washes with PBS, cells were blocked with 1% bovine serum albumin for 30 min. Samples were incubated overnight at 4°C with the relevant antibodies anti-MIF (1:500; Cell Signaling Technology) and anti-F actin (1:500; Santa-Cruz Biotechnology Inc.) for 2 h at room temperature. After three washes with PBS, immune-labeled cells were counterstained with 50 µl of DAPI at 37°C for 10 min. Cells were analyzed using a laser scanning confocal microscope (Leica Microsystems).
Sohn H.M., Kim B., Park M., Ko Y.J., Moon Y.H., Sun J.M., Jeong B.C., Kim Y.W, & Lim W. (2019). Effect of CD133 overexpression on bone metastasis in prostate cancer cell line LNCaP. Oncology Letters, 18(2), 1189-1198.
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