Lc3 antibody

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The LC3 antibody is a laboratory reagent used for the detection and analysis of the LC3 protein. LC3 is a key autophagy-related protein involved in the formation of autophagosomes. The antibody can be used in various immunoassay techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to study the expression and localization of LC3 in biological samples.

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Lab products found in correlation

Ecl plus chemiluminescent system, by Cytiva (1 mentions) Anti mouse caspase 1 p10, by Santa Cruz Biotechnology (1 mentions) P62 antibody, by Santa Cruz Biotechnology (1 mentions) Anti β actin antibody, by Santa Cruz Biotechnology (1 mentions) Anti 14 3 3 antibody, by Santa Cruz Biotechnology (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions)

3 protocols using lc3 antibody

Fluorescence Imaging of Autophagy Markers

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Cells grown on glass coverslips were treated with the test compounds for 24 hours. Next, slides were fixed with 4% paraformaldehyde for 20 minutes and treated with 0.05% Triton X‐100 for 15 minutes at RT. Thereafter, the coverslips were stained with LC3 antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) (diluted in 5% BSA) for 3 hours at RT, then washed with PBS thrice, followed by incubation with secondary antibodies at RT for 1 hour. Next, nuclei were stained with DAPI for 1 minute. Images were acquired using fluorescence microscopy.

Cui J., Man S., Cui N., Yang L., Guo Q., Ma L, & Gao W. (2018). The synergistic anticancer effect of formosanin C and polyphyllin VII based on caspase‐mediated cleavage of Beclin1 inhibiting autophagy and promoting apoptosis. Cell Proliferation, 52(1), e12520.

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Western Blot Analysis of Macrophage Signaling

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Aliquots of AMϕ lysates were separated on a 10% SDS-PAGE under non-reducing condition. Equivalent loading of the gel was determined by quantitation of protein as well as by reprobing membranes for actin detection. Separated proteins were electroblotted onto PVDF membrane and blocked for 1 h at room temperature with Tris-buffered saline containing 1% BSA. The membranes were then probed with primary antibody (polyclonal anti-NOD2, -NOD1, -IKKγ (phospho Ser31), -MIP-2, -MIF or −LC3 antibody purchased from Santa Cruz Biotechnology, Santa Cruz, CA) at room temperature for 1 h. After washing, primary antibodies associated with the membranes were detected on autoradiographic film by horseradish peroxidase-conjugated secondary antibodies and the ECL plus chemiluminescent system (Amersham, Arlington Heights, IL) according to the manufacturer's instructions. Blots were quantitated using Scion Image software (Scion Corp., Frederick, MD) and normalized to actin. Caspase-1 cleavage in the AMϕ was measured by detecting its p10 fragment in Western blot using rabbit polyclonal anti-mouse caspase-1 p10 (Santa Cruz Biotechnologies, CA).

Wen Z., Fan L., Li Y., Zou Z., Scott M.J., Xiao G., Li S., Billiar T.R., Wilson M.A., Shi X, & Fan J. (2014). Neutrophils Counteract Autophagy-Mediated Anti-Inflammatory Mechanisms in Alveolar Macrophage: Role in Post-Hemorrhagic Shock Acute Lung Inflammation. Journal of immunology (Baltimore, Md. : 1950), 193(9), 4623-4633.

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Evaluating Protein Expression in Rat Ventricle

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Western blots were performed as described previously9 (link). Briefly, the rats' left ventricle tissue and NRC samples were lysed in a lysis buffer containing 50 mM Tris, 150 mM NaCl, 1% Nonidet P-40, 0.25% superoxide dismutase, 1 mM EDTA, 1 mM NaF, 1 mM Na₃VO₃, 1 mM phenylmethylsulphonyl fluoride, and a proteinase inhibitor cocktail (Roche). The samples were evaluated by SDS-PAGE. Briefly, total protein from the samples was determined before being subjected to polyacrylamide gel electrophoresis and being transferred to a nitrocellulose (NC) membrane. The NC membrane was immunoblotted with an anti-14-3-3 antibody (Santa Cruz; 1:1,000), LC3 antibody (Santa Cruz; 1:1,000), p62 antibody (Santa Cruz; 1:1,000), and anti-β-actin antibody (Santa Cruz; 1:10,000). β-Actin protein served as a loading control. Protein bands were evaluated by densitometry using the Odyssey infrared imaging system (LI-COR).

Su F., Shi M., Zhang J., Zheng Q., Zhang D., Zhang W., Wang H, & Li X. (2018). Simvastatin Protects Heart from Pressure Overload Injury by Inhibiting Excessive Autophagy. International Journal of Medical Sciences, 15(13), 1508-1516.

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