Image lab quantification software

Manufactured by Bio-Rad

Sourced in United States

Image Lab is a quantification software developed by Bio-Rad. It provides tools for analyzing and quantifying data from a variety of imaging and analysis applications.

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Image Lab software (4753 citations) Image Lab (1121 citations)

6 protocols using image lab quantification software

Western Blot Analysis of Protein Expression

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30 μg of protein from whole cell lysates was resolved on 10% SDS-PAGE gels (Bio-Rad Laboratories, Hercules, CA) and transferred onto PVDF membranes (GE Healthcare, Piscataway, NJ). After transfer, membranes were blocked in 5% nonfat milk in TBST and membranes were incubated with specific antibodies (overnight at 4°C) followed by horseradish peroxidaseconjugated anti-mouse or anti-rabbit (Santa Cruz, Dallas, TX) immunoglobulin for 1 hour at RT. Signal was detected by Chemiluminescence Reagent (PerkinElmer, Waltham, MA) and visualized by autoradiography. Anti-human survivin antibody was purchased from R&D Systems (Minneapolis, MN). Anti-CUG-BP1, anti-HuR, and anti-GAPDH antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX). All primary anti-bodies were used at a dilution of 1:2000 and all secondary antibodies were used at a dilution of 1:4000. Signal intensity was quantified using Image Lab quantification software (Bio-Rad, Hercules, CA).

Phatak P., Byrnes K.A., Mansour D., Liu L., Cao S., Li R., Rao J.N., Turner D.J., Wang J.Y, & Donahue J.M. (2015). Overexpression of miR-214-3p in Esophageal Squamous Cancer Cells Enhances Sensitivity to Cisplatin by Targeting Survivin Directly and Indirectly Through CUG-BP1. Oncogene, 35(16), 2087-2097.

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Western Blot Analysis of Cell Signaling Proteins

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30 μg of protein from whole cell lysates was resolved on 10% SDS-PAGE gels (Bio-Rad Laboratories, Hercules, CA) and transferred onto PVDF membranes (GE Healthcare, Piscataway, NJ). After transfer, membranes were blocked in 5% nonfat milk in TBST and membranes were incubated with specific antibodies (overnight at 4°C) followed by horseradish peroxidase-conjugated anti-mouse or anti-rabbit (Santa Cruz, Dallas, TX) immunoglobulin for 1 hour at RT. Signal was detected by Chemiluminescence Reagent (PerkinElmer, Waltham, MA) and visualized by autoradiography. Anti-MAP3K11, anti-Cdc42, anti-c-Jun, anti-Rac-1, anti-cyclin D1 and anti-GAPDH were purchased from Santa Cruz Biotechnology (Dallas, TX). Anti-phosphorylated c-Jun (Ser62) II antibody was obtained from Cell Signaling Technology (Danvers, MA). Signal intensity was quantified using Image Lab quantification software (Bio-Rad, Hercules, CA).

Byrnes K.A., Phatak P., Mansour D., Xiao L., Zou T., Rao J.N., Turner D.J., Wang J.Y, & Donahue J.M. (2015). Overexpression of miR-199a-5p decreases esophageal cancer cell proliferation through repression of mitogen-activated protein kinase kinase kinase-11 (MAP3K11). Oncotarget, 7(8), 8756-8770.

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Quantitative Western Blot Analysis of RAB14

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Twenty micrograms of protein were resolved on 10% SDS-PAGE gels (Bio-Rad Laboratories, Hercules, CA, USA) and transferred onto PVDF membranes (GE Healthcare, Piscataway, NJ, USA). After transfer, membranes were blocked in 5% nonfat milk in TBST and incubated with anti-human RAB14 and anti-GAPDH antibodies (Santa Cruz, CA, USA) overnight at 4 °C followed by horseradish peroxidase-conjugated anti-mouse or anti-rabbit (Santa Cruz, Dallas, TX, USA) immunoglobulin secondary antibodies for 1 hour at room temperature. Signal was detected by Chemiluminescence Reagent (PerkinElmer, Waltham, MA) and visualized by autoradiography. Dilutions were 1:2000 for primary antibodies and 1:4000 for secondary antibodies. Signal intensity was quantified using Image Lab quantification software (Bio-Rad, Hercules, CA, USA). All the original blots are included as supplemental figures.

Phatak P., Burrows W.M., Creed T.M., Youssef M., Lee G, & Donahue J.M. (2022). MiR-214-3p targets Ras-related protein 14 (RAB14) to inhibit cellular migration and invasion in esophageal Cancer cells. BMC Cancer, 22, 1265.

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Western Blot Analysis of Protein Expression

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Immunoblot Analysis of PAK4 Expression

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Whole cell lysates were resolved on 10% SDS-PAGE gels (Bio-Rad Laboratories, Hercules, CA, USA) and Immunoblots were performed as reported earlier [4 (link)]. Anti-human PAK4 antibody was purchased from Cell Signaling (Danvers, MA, USA), anti-CD1 was purchased from Millipore (Billerica, MA, USA), anti-GAPDH, anti-tubulin, and horseradish peroxidase-conjugated anti-mouse or anti-rabbit antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX). Signal intensity was quantified using Image Lab quantification software (Bio-Rad, Hercules, CA, USA).

Phatak P., Burrows W.M., Chesnick I.E., Tulapurkar M.E., Rao J.N., Turner D.J., Hamburger A.W., Wang J.Y, & Donahue J.M. (2018). MiR-199a-3p decreases esophageal cancer cell proliferation by targeting p21 activated kinase 4. Oncotarget, 9(47), 28391-28407.

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Automated Western Blotting Protocol

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Whole cell lysates (20 μg) were resolved on 10% SDS-PAGE gels (Bio-Rad Laboratories, Hercules, CA, USA) and immunoblots were performed as reported earlier [12 (link)]. The automated Western system JESS was used to run lysates from si-Jun-B transfections. Anti-human Jun-B (1:2000) antibody was purchased from Proteintech (Danvers, MA, USA), anti-GAPDH (1:2000) and horseradish peroxidase-conjugated anti-rabbit antibodies (1:3000) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The signal intensity was quantified using Image Lab quantification software (Bio-Rad, Hercules, CA, USA) for conventional immunoblotting and a compass for simple Western software was used for automated Western.

Phatak P., Tulapurkar M.E., Burrows W.M, & Donahue J.M. (2023). MiR-199a-5p Decreases Esophageal Cancer Cell Proliferation Partially through Repression of Jun-B. Cancers, 15(19), 4811.

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