Ab75599

Manufactured by Abcam

Ab75599 is a laboratory product manufactured by Abcam. It serves as a research tool designed for scientific applications. The core function of this product is to provide a specific capability for use in experimental procedures. No further details can be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

3 protocols using ab75599

Evaluating EphrinB2 Signaling Pathway

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To reveal the mechanisms underlying EphrinB2-associated regulation, phosphorylation of key kinases in the EphB4/EphrinB2 reverse signalling pathway in BMSCs was evaluated following inducing increased levels of EphrinB2. The Western blot assay was performed as described above for EphrinB2. Briefly, membranes were incubated at 4 °C overnight with primary antibodies against intracellular PAK1 (PAK1; 1:1000 dilution; ab131522), phosphorylated PAK1 (p-PAK1; 1:1000 dilution; ab75599), Grb4 (1:1000 dilution; ab80620; all from Abcam) or GAPDH (1:1000 dilution; Santa Cruz Biotechnology). The membranes were then incubated with the corresponding secondary antibodies. The blots were developed using enhanced chemiluminescence Western blotting reagents and visualized using the Bio-Rad Image Lab system (Bio-Rad Laboratories).

Zhu M., Hua Y., Tang J., Zhao X., Zhang L, & Zhang Y. (2019). Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells. The Journal of International Medical Research, 47(7), 3282-3298.

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Protein Extraction and Western Blot Analysis

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Proteins were extracted from the cerebral cortex or cultured cells using the Membrane and Cytosol Protein Extraction Kit (Beyotime, China) and measured with the BCA Protein Assay Kit (Beyotime, China). Fifty micrograms protein per sample was separated through 10% ExpressPlus™ Gels (GenScript, the USA) and then transferred to Polyvinylidene fluoride (PVDF) membranes. After blocking with 5% skim milk in buffer, the membranes were incubated overnight at 4°C with the following antibodies: Caspase‐1 (89332, CST), Cleaved Caspase‐3 (ab214430, Abcam), COX‐2 (12282, CST), p‐JNK (ab124956, Abcam), JNK (ab179461, Abcam), p‐P38 (4511, CST), P38 (8690, CST), BAX (ab32504, Abcam), Bcl‐2 (ab182858, Abcam), MMP‐2 (87809S, CST), MMP‐9 (ab228402, Abcam), Catalase (ab76110, Abcam), Cu/Zn SOD (ab13498, Abcam), β‐actin (4970, CST), CD63 (EXOAB‐CD63A‐1, SBI), CD81 (EXOAB‐CD81A‐1, SBI), CD9 (EXOAB‐CD9A‐1, SBI), Rac1 (8631, CST), Cdc42 (8747, CST), PAK‐1(ab223849, Abcam) and p‐PAK1 (ab75599, Abcam). The membranes were washed and incubated with the secondary antibodies for 2 hours, and the positive bands were detected with a chromogenic developing agent.

Li H., Luo Y., Liu P., Liu P., Hua W., Zhang Y., Zhang L., Li Z., Xing P., Zhang Y., Hong B., Yang P, & Liu J. (2021). Exosomes containing miR‐451a is involved in the protective effect of cerebral ischemic preconditioning against cerebral ischemia and reperfusion injury. CNS Neuroscience & Therapeutics, 27(5), 564-576.

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Signaling Pathway Inhibitor Protocol

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Qu was purchased from Sigma-Aldrich (No. PHR1488; St Louis, MO). JSH-23, Go6983, and SCH772984 (specific inhibitors of NF-κb, PKC, and ERK1/2 respectively) and A-769662 (specific activator of AMPK) were purchased from Selleck (Houston, TX). Antibodies against uPA (ab131433), uPAR (ab82220), phosphor(p)-Pak1(ab75599), p-Limk1(ab38508), p-cofilin (ab12866), p-NF-κb (p65, S276) (ab106129), p-PKC-α (ab76016), p-PKC-β (ab75657), p-PKC-δ (ab133456), p-ERK1/2 (ab200807), and GAPDH (ab9485) that were used for western blotting were purchased by Abcam (Cambridge, England).

Li H, & Chen C. (2017). Quercetin Has Antimetastatic Effects on Gastric Cancer Cells via the Interruption of uPA/uPAR Function by Modulating NF-κb, PKC-δ, ERK1/2, and AMPKα. Integrative Cancer Therapies, 17(2), 511-523.

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