Profilin 1

Manufactured by Cell Signaling Technology

Profilin 1 is a protein that binds to actin monomers, regulating actin dynamics within the cell. It plays a crucial role in various cellular processes such as cell motility, endocytosis, and signal transduction.

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3 protocols using profilin 1

Lung Cancer Tissue Immunofluorescence Analysis

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Human lung cancer tissue slides were obtained from the Lung Cancer Biospecimen Research Network (LCBRN), University of Virginia, Charlottesville, VA. Normal (n = 4) and malignant (n = 4) lung tissue slides were processed for immunofluorescence labeling using standard protocols (45 (link)). Tissue sections were stained with profilin 1, cofilin 1, VASP, and VASP^pS239 (dilution, 1: 500) (Cell Signaling Technology, Inc., Danvers, MA) and anti-human rabbit monoclonal antibodies targeting VASP^pS157 (dilution, 1: 500) (Santa Cruz, Dallas, TX) followed by secondary antibodies Alexa 488 labeled IgG (1:1000) and nuclei were stained with DAPI (Invitrogen, Carlsbad, CA). Images (4 from each slide) were taken using Carl Zeiss’s Axio Scope A1 Polarized Light Fluorescent Microscope (Carl Zeiss, Jena, Germany) with 400X magnification and identical settings. Intensity of image color was quantified using NIH Image J software using identical background settings. The results expressed as intensity/cell.

Ali M., Heyob K., Jacob N.K, & Rogers L.K. (2016). Alterative Expression and Localization of Profilin 1/VASPpS157 and Cofilin 1/VASPpS239 Regulates Metastatic Growth and is Modified by DHA Supplementation. Molecular cancer therapeutics, 15(9), 2220-2231.

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Comprehensive Antibody Validation for Cellular Analysis

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Primary antibodies against following proteins were used: CAP1 (Abcam ab133655), CD36 (Santa Cruz H-300 sc-9154), Syk (Santa Cruz 4D10 sc-1240), GAPDH (Calbiochem 6C5-CB1001), β-actin (Abcam ab20272), β3-integrin (Santa Cruz HC93 sc-14009), cofilin (Cell Signaling Technology D3F9 #5175), profilin-1 (Cell Signaling Technology #3237), and vinculin (Sigma SAB4200080). Secondary antibodies Alexa Fluor 568- or 488-conjugated anti-mouse and anti-rabbit immunoglobulins (Molecular Probes, Invitrogen Life Technologies Ltd.) were used for immunofluorescence. Peroxidase-conjugated anti-mouse and anti-rabbit immunoglobulins (Sigma-Aldrich Co. Ltd.) or IRDye 680 or IRDye 800 anti-mouse and anti-rabbit immunoglobulins (LI-COR Biosciences, Lincoln, USA) were used for Western blot.
Human fibrinogen was from Enzyme Research (Swansea, UK), collagen (Kollagenreagens Horm) was from Takeda (Osaka, Japan), recombinant human resistin (450-19) was from PeproTech (London, UK), latrunculin B and GSNO were from Enzo Life Sciences (Exeter, UK). PGI2 was from Cayman Chemical (Michigan, USA). Glycogen synthase kinase 3 (GSK3) inhibitor CHIR99021 was from Abcam. Other reagents were from Sigma-Aldrich unless otherwise indicated.

Joshi P., Riley D.R., Khalil J.S., Xiong H., Ji W, & Rivero F. (2018). The membrane-associated fraction of cyclase associate protein 1 translocates to the cytosol upon platelet stimulation. Scientific Reports, 8, 10804.

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Antibodies for Western Blotting and Flow Cytometry

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Antibodies for Western blotting were purchased from the following sources: Histone H2A (Cell Signaling Technology #3636); HMGB1 (Cell Signaling Technology #6893); β-Actin (Cell Signaling Technology #3700); Profilin-1 (Cell Signaling Technology #3237); Enolase-1 (Cell Signaling Technology #3810); Bip/GRP78 (Cell Signaling Technology #3177); Alpha-fodrin (EMD Millipore #MAB1622); MLKL (Cell Signaling Technology #14993); Phospho-MLKL Ser358 (Cell Signaling Technology #91689); GAPDH (ThermoFisher #AM4300). Antibodies for flow cytometry were purchased from the following sources: F4/80, PE-Cyanine5 (ThermoFisher #15-4801-82); CD11b APC-eFluor 780 (ThermoFisher #47-0112-82); CD11c Alexa Fluor 700 (ThermoFisher #56-0114-82); CD86 (B7-2) FITC (ThermoFisher #11-0862-82); CD40 Super Bright 436 (ThermoFisher #62-0401-82); MHC Class II I-Ab (ThermoFisher #17-5320-82); CD25 Alexa Fluor 488 (ThermoFisher #53-0251-82); CD69 Alexa Fluor 700 (BioLegend #104539).

Andersohn A., Garcia M.I., Fan Y., Thompson M.C., Akimzhanov A.M., Abdullahi A., Jeschke M.G, & Boehning D. (2019). Aggregated and Hyperstable Damage-Associated Molecular Patterns Are Released During ER Stress to Modulate Immune Function. Frontiers in Cell and Developmental Biology, 7, 198.

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