Ab9722

Manufactured by Cell Signaling Technology

Sourced in United Kingdom

Ab9722 is a primary antibody product offered by Cell Signaling Technology. It is designed to detect specific target proteins in various applications, such as Western blotting and immunohistochemistry. The core function of Ab9722 is to provide a reliable and specific tool for researchers to identify and study their protein of interest.

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Lab products found in correlation

3 protocols using ab9722

Western Blot Analysis of Cardiac Markers

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The heart tissue and H9C2 cells were ground and lysed by protein lysis buffer (Beyotime, China) with PMSF and phosphatase inhibitor cocktail. Total protein samples were extracted from the lysate, and the protein concentration was detected. The sample was loaded into 8% or 12% SDS-PAGE gel for electrophoresis and then transferred to the PVDF membrane. It was blocked for 1 hour and then immersed in primary antibody for 12 hours at 4°C, such as GAPDH (Proteintech, 60004-1-Ig, USA), GSDMD (Abcam, ab219800, ab209845, UK), NLRP3 (Abcam, ab214185, UK), caspase-1 (Abcam, ab179515, UK), IL-1β (Abcam, ab9722, UK), caspase-3 (Cell Signaling Technology, 14220T, USA), caspase-7 (Cell Signaling Technology, 12827T), caspase-8 (Beyotime, AC056, China), Bcl-2 (Beyotime, AB112, China), Bax (Beyotime, AB026, China), PARP-1 (Cell Signaling Technology, 9532S, USA), and poly/mono-ADP ribose (Cell Signaling Technology, 83732S, USA). The membranes were washed with TBST twice, immersed in the secondary antibody for 1 hour, and then washed again. Bands were displayed by the ECL detection kit (Millipore, USA).

Zhang Z.H., Zhang Z.G., Chen M.W., Yang Y., Li R.J., Xu J.J., Yang C., Li Y.Y., Chen H.W., Liu S.X., Li Y.L., Luo P., Liu Y.J., Chen W.B., Shan Z.G, & Huang Z.R. (2022). Inhibition of GSDMD Activates Poly(ADP-ribosyl)ation and Promotes Myocardial Ischemia-Reperfusion Injury. Oxidative Medicine and Cellular Longevity, 2022, 1115749.

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Western Blot Analysis of TLR4, NF-κB Pathway

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Proteins were extracted from treated HUVECs and ApoE^−/− mice aortas. All samples were lysed on ice for 30 min in a RIPA buffer (Beyotime, Nantong, China) after being ground with a pestle. Equal amounts of protein were separated by SDS-PAGE and transferred to PVDF membranes. After blocking by 5% non-fat milk, the membranes were incubated with primary antibodies for TLR4 (Abcam, ab120684-1, 1:500), p65 (Abcam, ab7970, 1:1000), interleukin-1β (IL-1β)(Abcam, ab9722, 1:500) and p-p65 (Ser536) (Cell signaling, #3033, 1:1000) overnight at 4 °C. After washing and incubating with an HRP-conjugated secondary antibody (1:10,000), the immunoreactive bands were visualized using chemiluminescence (Millipore Corporation, Billerica, MA, USA). Sample loadings were normalized to β-actin expression.

Zeng X., Guo R., Dong M., Zheng J., Lin H, & Lu H. (2018). Contribution of TLR4 signaling in intermittent hypoxia-mediated atherosclerosis progression. Journal of Translational Medicine, 16, 106.

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Astrocyte Polarization Induction and Targeting

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A1 reactive astrocytes were generated by culturing the purified astrocytes on PDMS coated tissue culture plates and then treating for 24 h with IL-1α (3 ng/ml, Sigma, I3901), TNF-α (30 ng/ml, Cell Signaling Technology, 8902SF), and C1q (400 ng/ml, MyBioSource, MBS143105). A2 reactive astrocytes were generated by culturing the purified astrocytes on PDMS coated tissue culture plates and then treating for 24 h with IL-1β (30 ng/ml, Cell Signaling Technology, 8900SF) and TNF-α (30 ng/ml, Cell Signaling Technology, 8902SF). A1 reactive astrocytes were targeted for 48 h using neutralizing antibodies to IL-1α (30 ng/ml, Abcam, ab9614), TNF-α (30 ng/ml, Cell Signaling Technology, 7321), and TGF-β (30 ng/ml, R&D Systems, 243-B3-002/CF). A2 reactive astrocytes were targeted for 48 h using neutralizing antibodies to IL-1β (30 ng/ml, Abcam, ab9722), TNF-α (30 ng/ml, Cell Signaling Technology, 7321), and IL-6 (30 ng/ml, Abcam, ab6672) [36] , [37] . Polydimethylsiloxane (PDMS) coated tissue culture plates were prepared by mixing Sylgard-184 elastomer and curing agents at a ratio of 10:1 (w/v), then pouring into the plates and curing for 48 h.

Khodadadei F., Arshad R., Morales D.M., Gluski J., Marupudi N.I., McAllister J.P., Limbrick D.D., & Harris C.A. (2021). The Effect of A1/A2 Reactive Astrocyte Expression on Hydrocephalus Shunt Failure.

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