Mab835

Manufactured by R&D Systems

Sourced in United States

MAB835 is a monoclonal antibody that recognizes human PCSK9. It can be used for the detection and quantification of PCSK9 in various sample types.

Automatically generated - may contain errors

Lab products found in correlation

Ap106p, by Merck Group (1 mentions) Image labtm software version 5, by Bio-Rad (1 mentions) Polyvinylidene difluoride membrane, by Merck Group (1 mentions) Chemidoc xrs system, by Bio-Rad (1 mentions) Vcx 500 sonicator, by Sonics (1 mentions) Af375, by R&D Systems (1 mentions) Supersignal west femto chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Gel doc imager, by Bio-Rad (1 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Anti rabbit igg hrp, by Cell Signaling Technology (1 mentions) Cdc20 10252 1 ap, by Proteintech (1 mentions) γh2ax, by Cell Signaling Technology (1 mentions) Puma 55120 1 ap, by Proteintech (1 mentions) Mcl 1, by Cell Signaling Technology (1 mentions) Ab133534, by Abcam (1 mentions) Ab234436, by Abcam (1 mentions) P chk1, by Cell Signaling Technology (1 mentions) Bax 50599 2 ig, by Proteintech (1 mentions) Caspase 7, by Cell Signaling Technology (1 mentions)

3 protocols using mab835

Western Blot Analysis of Liver Tumor Organoids

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Liver tumor organoids were lysed using sample buffer [62.5 mM Tris-HCl, pH 6.8, 34.7 mM sodium dodecyl sulfate (SDS), 5% β-mercaptoethanol, and 10% glycerol], sonicated, boiled for 5 min, subjected to SDS-polyacrylamide gel electrophoresis, and transferred onto a polyvinylidene difluoride membrane (Millipore). Sonication was performed using a VCX 500 sonicator (Sonics, Newtown, CT, USA) with 20% amplitude for 4 s in a 1 s-on/1 s-off mode. The membrane was blocked with 5% skim milk in Tris-HCl-buffered saline containing 0.05% Tween 20 (TBST) for 30 min and incubated overnight at 4 °C with primary antibodies against TRAIL (AF375; R&D Systems, Minneapolis, MA, USA), cleaved caspase-3 (CC3, MAB835; R&D Systems), caspase-3 (pro-C3, MAB707; R&D Systems), and GAPDH (sc47724; Santa Cruz Biotechnology, Santa Cruz, CA, USA). The membrane was washed thrice for 5 min with TBST and incubated with horseradish peroxidase-conjugated secondary antibodies (7074S and 7076S; Cell signaling Technology Danvers, MA, USA, or AP106P; Millipore) for 1 h. After washing thrice with TBST, the bands were visualized using EZ-Western Lumi Pico or Femto (Dogen, Seoul, Republic of Korea) and recorded using a ChemiDoc XRS+ System with Image Lab^TM software Version 5.2.1 (Bio-Rad Laboratories, Hercules, CA, USA).

Yoon Y., Kim C.W., Kim M.Y., Baik S.K., Jung P.Y, & Eom Y.W. (2024). Interferon-β Overexpression in Adipose Tissue-Derived Stem Cells Induces HepG2 and Macrophage Cell Death in Liver Tumor Organoids via Induction of TNF-Related Apoptosis-Inducing Ligand Expression. International Journal of Molecular Sciences, 25(2), 1325.

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NLGN3 Shedding in Optic Nerve Lysates

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Optic nerves were lysed in RIPA buffer (Santa Cruz Biotechnology sc-24948) supplemented with 1× phosphatase inhibitor (Thermo Fisher Scientific 78428) and 1× protease inhibitor (Thermo Fisher Scientific 87786). Protein concentrations were determined using the Pierce BCA protein assay kit (Thermo Fisher Scientific 23225). Twenty μg of optic nerve lysate or 5 μg of conditioned medium was used for SDS–PAGE. PVDF membranes were subsequently blocked in 5% BSA and Tris buffered saline with 1% Triton X-100 (TBST), followed by primary antibody (rabbit anti-NLGN3, 1:500, Novus NBP1-90080 lot B118108; rabbit anti-β-actin, 1:10,000, Cell Signaling 4970 lot 15; rabbit anti-ADAM10, 1:250, Abcam ab124695 lot GR3244660-5; rabbit anti-cleaved caspase-3, 1:500, R&D Systems MAB835 lot KHK051810A; and rabbit anti-caspase-3, 1:1,000, Cell Signaling 9662 lot 19) incubation overnight at 4 °C and secondary antibody (1:10,000 anti-rabbit IgG-HRP, Cell Signaling 7074) incubation for 1 h at room temperature. Blots were developed using the SuperSignal West Femto Chemiluminescent Substrate (Thermo Fisher Scientific 34095), imaged on a Bio-Rad Gel Doc imager using Image Lab software (v.6) and analysed using Image Studio Lite (v.5.0.21). The intense overexposed full-length NLGN3 band was cropped out to allow for easy identification of shed NLGN3.

Pan Y., Hysinger J.D., Barron T., Schindler N.F., Cobb O., Guo X., Yalçın B., Anastasaki C., Mulinyawe S.B., Ponnuswami A., Scheaffer S., Ma Y., Chang K.C., Xia X., Toonen J.A., Lennon J.J., Gibson E.M., Huguenard J.R., Liau L.M., Goldberg J.L., Monje M, & Gutmann D.H. (2021). NF1 mutation drives neuronal activity-dependent initiation of optic glioma. Nature, 594(7862), 277-282.

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Western Blot Antibody Protocol

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Western blot was conducted as previously described [35 (link)]. The antibodies for Western blotting were as follows: Mcl-1 (#5453, 1:1000, Cell Signaling Technology, Danvers, MA, USA), p-Chk1 (#2348, 1:1000, Cell Signaling Technology, Danvers, MA, USA), caspase-7 (#9492S, 1:1000, Cell Signaling Technology, Danvers, MA, USA), Bak (#3814, 1:1000, Cell Signaling Technology, Danvers, MA, USA), γH2AX (#9718, 1:1000, Cell Signaling Technology, Danvers, MA, USA), CDC20 (10252-1-AP, 1:1000, Proteintech, Rosemont, IL, USA), Bax (50599-2-Ig, 1:1000, Proteintech, Rosemont, IL, USA), Puma (55120-1-Ap, 1:1000, Proteintech, Rosemont, IL, USA), Bcl-2 (12789-1-Ap, 1:1000, Proteintech, Rosemont, IL, USA), Bcl-xL (10783-1-Ap, 1:1000, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (MAB835, 1:1000, R&D Systems, Minneapolis, MN, USA), cleaved caspase-9 (sc8355, 1:1000, Santa Cruz, CA, USA), Rad51 (ab133534, 1:1000, Abcam, Cambridge, UK), apaf1(ab234436, 1:1000, Abcam, Cambridge, UK) and β-actin (T0022,1:3000, Affinity, San Francisco, CA, USA).

Gao Y., Wen P., Chen B., Hu G., Wu L., Xu A, & Zhao G. (2020). Downregulation of CDC20 Increases Radiosensitivity through Mcl-1/p-Chk1-Mediated DNA Damage and Apoptosis in Tumor Cells. International Journal of Molecular Sciences, 21(18), 6692.

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