Ab105767

Manufactured by Abcam

Ab105767 is a laboratory equipment product manufactured by Abcam. It serves as a core function for specific laboratory applications. A detailed description of the product's intended use or performance characteristics is not available at this time.

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Lab products found in correlation

Sc 7292, by Santa Cruz Biotechnology (2 mentions) Sc 53540, by Santa Cruz Biotechnology (2 mentions) Ab62344, by Abcam (2 mentions) Ab39670, by Abcam (2 mentions) Ab208910, by Abcam (2 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) Mg132, by Merck Group (1 mentions) Bafilomycin a1, by Merck Group (1 mentions) Anti flag, by Merck Group (1 mentions) Z vad fmk, by Apexbio (1 mentions) Anti actin sc 1616, by Santa Cruz Biotechnology (1 mentions) Mtnfα, by Thermo Fisher Scientific (1 mentions) Ab187091, by Abcam (1 mentions) Ab 33174, by Abcam (1 mentions) Mifn β, by PBL Assay Science (1 mentions) Htnfα, by Thermo Fisher Scientific (1 mentions) Lcl161, by Chemietek (1 mentions) Nbp1 77299, by Novus Biologicals (1 mentions) Bapta am, by Bio-Techne (1 mentions)

5 protocols using ab105767

Molecular Mechanisms of Cell Death

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The dimerizer (B/B) and its competitor (washout) were purchased from Clontech. The cytokines used were: mTNFα (Peprotech), mIFNβ (PBL Assay Science) and hTNFα (Peprotech). The SMAC mimetic and inhibitor of IAP (LCL-161) was purchased from Chemietek. Other reagents included CHX (C7698, sigma), Q-VD-OPh (qVD, A8165, Apexbio), 7-Cl-O-Nec-1 (Nec-1s, 504297, calbiochem), zVAD-fmk (A1902, Apexbio), MG132 (Sigma), Necrosulfonamide (NSA, calbiochem), LPS (Sigma), BAPTA-AM (Tocris) and Bafilomycin A1 (Sigma). Antibodies used for immunoblotting in this study were anti-actin (sc-1616, Santa Cruz) anti-FLAG (A8592, Sigma), anti-HA (H6908, Sigma), anti-hMLKL (M6697, Sigma), anti-mMLKL (AP14272b, Agent), anti-pMLKL (ab187091, Abcam), anti-RIPK3 (NBP1-77299, Novus), anti-ALIX (3A9, 2171S, Cell signaling Technology), anti-CHMP2B (ab33174, abcam), anti-CHMP4B (ab105767, abcam), anti-TSG101 (4A10, ab83, abcam) anti-TNF (Clone XT3.11 from BioXCell) and anti-IκBα (C-21, Santa Cruz).

Gong Y.N., Guy C., Olauson H., Becker J.U., Yang M., Fitzgerald P., Linkermann A, & Green D.R. (2017). ESCRT-III acts downstream of MLKL to regulate necroptotic cell death and its consequences. Cell, 169(2), 286-300.e16.

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Antibody Characterization for Cell Imaging

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These studies used commercial mouse monoclonal antibodies against ALIX (sc-53540; Santa Cruz Biotechnology, Dallas, TX), α-tubulin (DM1A; Sigma, St. Louis, MO), lamin A/C (sc-7292; Santa Cruz Biotechnology); commercial rabbit polyclonal antibody against GFP (598; MBL, Nagoya, Japan), TagRFP (AB233; Evrogen, Moscow, Russia), CHMP4B (ab105767; Abcam, Cambridge, UK).

Arii J., Maeda F., Maruzuru Y., Koyanagi N., Kato A., Mori Y, & Kawaguchi Y. (2020). ESCRT-III controls nuclear envelope deformation induced by progerin. Scientific Reports, 10, 18877.

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Protein Extraction and Analysis from Trauma Site

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Cytosolic, cytomembrane and overall extracts of protein were requested following the previous description.²⁸, ²⁹ Proteins at the site of trauma and on the contralateral side were harvested at different time and extracted using radioimmunoprecipitation assay (RIPA) buffer on ice (Sigma) and then centrifuged for 15 minutes 1000 g 4°C. The proteins underwent separation using 12% SDS‐PAGE gel and placed to polyvinylidene fluoride (PVDF) membranes (Millipore). Membranes underwent incubation with 5% (w/v) non‐fat dried milk for 2 hours at ambient temperatures and then incubated using primary antibodies throughout the night at 4°C. The membranes underwent incubation in the antibodies below: GAPDH (1:2000, #5174; Cell Signaling Technology), RIP3 (1 µg/mL, ab62344; Abcam), p‐MLKL (1:500, ab208910, Abcam), CHMP4B (1 µg/mL, ab105767; Abcam) and FOXO1 (1:1000, ab39670; Abcam). Using an enhanced chemiluminescence detection system (GE Healthcare), we ascertained bound antibodies. Applying ImageJ software (National Institutes of Health), we studied the achieved bands’ optical density.

Zhao P., Li C., Chen B., Sun G., Chao H., Tu Y., Bao Z., Fan L., Du X, & Ji J. (2020). Up‐regulation of CHMP4B alleviates microglial necroptosis induced by traumatic brain injury. Journal of Cellular and Molecular Medicine, 24(15), 8466-8479.

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Antibody Sources and Dilutions for Research

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Antibodies were purchased and used as follows: commercial mouse monoclonal antibodies against ALIX (sc-53540; Santa Cruz Biotechnology, Dallas, TX; 1:500), Flag (M2, F3165; Sigma, St. Louis, MO; 1:1000), α-tubulin (DM1A, T9026; Sigma; 1:1000), lamin A/C (sc-7292; Santa Cruz Biotechnology; 1:2000), Emerin (4G5, MS-1751-S; Lab Vision Corporation, Fremont, CA; 1:1000), gB (P1105; Virusys Corporation, Taneytown, MD; 1:2000), Strep-tag (4F1, M211-3; MBL, Nagoya, Japan; 1:1000), and DFz2 (12A7; DSHB, Iowa City, IA; 1:6); commercial rabbit polyclonal antibody against GFP (598; MBL; 1:1000), CHMP4B (ab105767; Abcam, Cambridge, UK; 1:1000), CHMP4C (ab155668; Abcam; 1:500), lamin B1 (ab16048-100; Abcam; 1:1000) and calnexin (c4731; Sigma; 1:2000); and commercial mouse polyclonal antibodies against CHMP4A (ab67058; Abcam; 1:500). Mouse polyclonal antibodies against UL31(1:500) and rabbit polyclonal antibodies against UL34 (1:2000) were reported previously^{61 (link)}.

Arii J., Watanabe M., Maeda F., Tokai-Nishizumi N., Chihara T., Miura M., Maruzuru Y., Koyanagi N., Kato A, & Kawaguchi Y. (2018). ESCRT-III mediates budding across the inner nuclear membrane and regulates its integrity. Nature Communications, 9, 3379.

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Immunohistochemical Analysis of Cell Death

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The frozen brain sections were permeabilized for 20 minutes with 0.2% Triton X‐100 (Sigma‐Aldrich, St Louis, MO; USA, X100); then, they underwent blocking processed with 5% normal goat serum (Millipore; S26‐LITER) and incubation throughout the night using major antibodies and then using minor antibodies for 2 hours at ambient temperatures. Nuclei underwent staining processed using DAPI. Major antibodies included RIP3 (5 µg/mL, ab62344; Abcam), p‐MLKL (1:100, ab208910, Abcam), CHMP4B (5 µg/mL, ab105767; Abcam) and FOXO1 (1‐5 µg/mL, ab39670; Abcam). Cell processing was like sections, except that they underwent initial fixing processed for 20 minutes with pre‐cooled paraformaldehyde (4%, w/v). Using an Olympus FluoView confocal microscope with appropriate emission filters (Olympus), we observed immunoreactivity.

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