Western Blot Analysis of Key Proteins

Protein extracts were separated and transferred to a polyvinylidene fluoride membrane. First, the primary antibody was added to the membrane for overnight incubation at 37 °C. Then, horseradish peroxidase-labeled secondary antibody (ab205718, goat anti-rabbit or goat anti-mouse, 1:10,000, Abcam) was added for further incubation followed by visualization using enhanced chemiluminescence (Shanghai Baoman Biotechnology, Shanghai, China). The primary antibodies procured from Abcam (CA, UK) included KLF4 (ab215036, 1:1000, rabbit; ab214666, 1:1000, rabbit), α‑SMA (ab32575, 1:1000–1:5000, rabbit), PCNA (ab92552, 1:1000–1:10000, rabbit), MMP2 (ab92536, 1:1000–1:5000, rabbit), MMP3 (ab52915, 1:1000–1:20000, rabbit), MMP9 (ab76003, 1:1000–1:20000, rabbit), p-p38 (ab4822, 1:1000, rabbit), p38 (ab170099, 1:1000–1:5000, rabbit). In addition, GAPDH (ab181602, 1:10,000, rabbit, Abcam) was used as an internal reference, and the gel image analysis software was used for quantitative analysis.⁴⁶ (link)

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Huang Z., Hu J., Xu J., Wang H, & Dai L. (2023). microRNA-130b May Induce Cerebral Vasospasm after Subarachnoid Hemorrhage via Modulating Kruppel-like Factor 4. Molecular and Cellular Biology, 43(7), 301-316.

Publication 2023

ab205718 goat anti-mouse ab215036 ab214666 ab32575 ab92552 ab92536 ab52915 ab76003 ab4822 ab170099 ab181602

Antibodies Antibody Chemiluminescence Gapdh Goat Horseradish peroxidase Klf4 Membrane Mmp2 Mmp3 Mmp9 Mouse Pcna Polyvinylidene fluoride Protein Rabbit

Corresponding Organization :

Other organizations : Southern University of Science and Technology, Jinan University

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Variable analysis

independent variables

Primary antibody incubation at 37 °C overnight
Horseradish peroxidase-labeled secondary antibody incubation

dependent variables

Protein expression levels of KLF4, α-SMA, PCNA, MMP2, MMP3, MMP9, p-p38, p38

control variables

GAPDH as an internal reference

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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