Right lungs were collected at the time of euthanasia and snap frozen. Proteins were isolated from tissue homogenates in RIPA buffer (Boston Bioproducts, Ashland, MA, USA) with protease inhibitors26 . The protein concentration in cell lysates was measured using the BCA Assay (Pierce, Thermo Scientific). 20 μg of proteins were separated on a 10–20% Tris-Glycine gel (Novex, Life Technologies) and transferred to a PVDF membrane. After blocking overnight at 4°C in 5% milk, blots were incubated with a goat polyclonal IL-33 (1:500, R&D Systems) or mouse monoclonal β-actin (1:1000, Cell Signaling, Danvers, MA) antibodies diluted in TBST at RT for 2h, followed by a rabbit anti-goat or goat anti-mouse secondary antibody (1:3000, BioRad) diluted in TBST for 1h at RT. The blots were visualized using the Supersignal West Femto Chemiluminescent substrate (Thermo Scientific) and imaged by a KODAK M35A X-OMAT processor.
Rabbit anti goat or goat anti mouse secondary antibody
Manufactured by Bio-Rad
Rabbit anti-goat or goat anti-mouse secondary antibody is a laboratory reagent used to detect the presence of primary antibodies raised in goat or mouse, respectively. It functions by binding to the Fc region of the primary antibody, allowing for the detection and visualization of the target antigen.
Automatically generated - may contain errors
Lab products found in correlation
Goat polyclonal il 33, by R&D Systems (2 mentions)
M35 a x omat processor, by Kodak (2 mentions)
Supersignal west femto chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions)
Ripa buffer, by Boston BioProducts (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
Bca assay, by Thermo Fisher Scientific (2 mentions)
2 protocols using rabbit anti goat or goat anti mouse secondary antibody
1
Western Blot Analysis of IL-33 Expression in Lung Tissue
2
Western Blot Analysis of IL-33 Expression in Lung Tissue
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Right lungs were collected at the time of euthanasia and snap frozen. Proteins were isolated from tissue homogenates in RIPA buffer (Boston Bioproducts, Ashland, MA, USA) with protease inhibitors26 . The protein concentration in cell lysates was measured using the BCA Assay (Pierce, Thermo Scientific). 20 μg of proteins were separated on a 10–20% Tris-Glycine gel (Novex, Life Technologies) and transferred to a PVDF membrane. After blocking overnight at 4°C in 5% milk, blots were incubated with a goat polyclonal IL-33 (1:500, R&D Systems) or mouse monoclonal β-actin (1:1000, Cell Signaling, Danvers, MA) antibodies diluted in TBST at RT for 2h, followed by a rabbit anti-goat or goat anti-mouse secondary antibody (1:3000, BioRad) diluted in TBST for 1h at RT. The blots were visualized using the Supersignal West Femto Chemiluminescent substrate (Thermo Scientific) and imaged by a KODAK M35A X-OMAT processor.
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