Cells were seeded on Matrigel (Growth Factor Reduced Matrigel, BD Biosciences, Franklin Lakes, NJ, USA) using 2% Matrigel DMEM media supplemented with 10% fetal bovine serum and penicillin/streptomycin. Immunostaining with cell-based organoids was performed following a previously published protocol.36 (link) Primary antibodies used were anti-E-cadherin (BD Bioscience, 610181) and anti-F-actin (Thermo Fisher, A12381). Images were acquired using a Zeiss LSM 510 microscope and analyzed with ImageJ software.
Anti f actin
Manufactured by Thermo Fisher Scientific
Sourced in United States
Anti-F-actin is a primary antibody that specifically recognizes and binds to F-actin, the filamentous form of actin, a key cytoskeletal protein found in eukaryotic cells. This antibody can be used in various immunodetection techniques to visualize and study the distribution and dynamics of the actin cytoskeleton.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 510 microscope, by Zeiss (1 mentions)
Anti e cadherin, by BD (1 mentions)
Growth factor reduced matrigel, by BD (1 mentions)
Anti v5, by Thermo Fisher Scientific (1 mentions)
K02288, by Selleck Chemicals (1 mentions)
Anti ki67, by Abcam (1 mentions)
Anti cleaved caspase 3, by Thermo Fisher Scientific (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Anti β tubulin, by Abcam (1 mentions)
Anti egfr, by Abcam (1 mentions)
Repsox, by Merck Group (1 mentions)
Anti smad2 3, by Santa Cruz Biotechnology (1 mentions)
Anti p smad2 3, by Santa Cruz Biotechnology (1 mentions)
Anti pegfr y1068, by Abcam (1 mentions)
Galunisertib ly2157299, by Selleck Chemicals (1 mentions)
A83 01, by MedChemExpress (1 mentions)
Anti hepsin, by R&D Systems (1 mentions)
Anti ph3, by Santa Cruz Biotechnology (1 mentions)
Anti smad2, by Santa Cruz Biotechnology (1 mentions)
2 protocols using anti f actin
1
Immunostaining of Cell-Based Organoids
2
Inhibitors and Antibodies for Cellular Signaling
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The inhibitors used in this study were K02288 (Selleckchem, S7359, Houston, TX, USA), Galunisertib/LY2157299 (Selleckchem, S2230), RepSox (Sigma‐Aldrich/Merck, R0158), A‐83‐01 (MedChem Express, HY‐10432, Monmouth Junction, NJ, USA), Erlotinib (EGFRi, Selleckchem, S1023), and human ALK1 inhibitory antibody (R&D Systems, MAB3701). The hepsin inhibitory antibody (Ab25) was a kind gift from Dr. Kirchhofer, Genentech [10 (link)]. ZFH7116 provided by Dr. Janetka [12 (link)]. The antibodies used in this study were Anti‐sheep HRP (Upstate cell signaling solutions, #12‐342), anti‐rabbit HRP (Millipore, AP132P, Burlington, MA, USA), anti‐hepsin (R&D Systems, AF4776), anti‐GAPDH (CST#2118S), anti‐β‐tubulin (Abcam, ab6046, Cambridge, UK), anti‐pEGFR Y1068 (Abcam, ab40815), anti‐EGFR (Abcam, ab32077), anti‐pSmad2/3 (CST#8828), anti‐Smad2/3 (CST#8685), anti‐Smad2 (CST#5339), anti‐pH3 (Santa Cruz, sc‐8656, Dallas, TX, USA), total‐H3 (CST#9715), anti‐V5 (Invitrogen, #R96025), anti‐Ki67 (Abcam, ab15580), anti‐cleaved caspase 3 (CST#9661), anti‐F‐actin (phalloidin staining) (Thermo Fisher Scientific, A22283), anti‐pSmad1/5 (CST#9516) and anti‐ALK‐1 (R&D Systems, AF370).
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