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Mouse hrp

Manufactured by Santa Cruz Biotechnology
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Mouse-HRP is a secondary antibody conjugated with Horseradish Peroxidase (HRP). It is designed for the detection and visualization of mouse primary antibodies in various immunoassays, such as Western blotting, Enzyme-Linked Immunosorbent Assay (ELISA), and immunohistochemistry.

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Lab products found in correlation

α tubulin, by Merck Group (2 mentions) Rabbit hrp, by Santa Cruz Biotechnology (2 mentions) Rabbit hrp, by Jackson ImmunoResearch (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions) Flag tag (flag), by Abcam (1 mentions) Gadph, by Santa Cruz Biotechnology (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Ab3691, by Abcam (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) β actin, by Merck Group (1 mentions) Rabbit hrp, by Abcam (1 mentions) U0126, by Santa Cruz Biotechnology (1 mentions) Lcn2 antibody, by R&D Systems (1 mentions) Phosphorylated erk p erk, by Cell Signaling Technology (1 mentions) Matrigel matrix, by Corning (1 mentions) Rad52, by Thermo Fisher Scientific (1 mentions) Pkap1, by Fortis Life Sciences (1 mentions) Mcl 1, by Santa Cruz Biotechnology (1 mentions) Primary antibodies against cleaved caspase 3, by Cell Signaling Technology (1 mentions)

8 protocols using mouse hrp

1

Protein Analysis in Embryos

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Total protein was extracted from embryos and analyzed on 10% SDS-PAGE followed by western blot analysis according to the procedures described by Lin et al. [16 (link)], except that the yolk was removed by deyolking buffer (55 mM NaCl, 1.8 mM KCl and 1.25 mM NaHCO3) and antibodies against Flag (Abcam, Cambridge, UK; 1:5000 dilution), α-tubulin (Sigma-Aldrich, St. Louis, MO, USA; 1:5000 dilution), mouse-HRP (Santa Cruz, Santa Cruz, CA, USA; 1:5000 dilution), and rabbit-HRP (Santa Cruz; 1:5000 dilution) were used.
Lin C.Y., Lin H.Y., Chuang C.K., Zhang P.H., Tu R.Y., Lin S.P, & Tsai H.J. (2020). Effect of Mutated ids Overexpression on IDS Enzyme Activity and Developmental Phenotypes in Zebrafish Embryos: A Valuable Index for Assessing Critical Point-Mutations Associated with Mucopolysaccharidosis Type II Occurrence in Humans. Diagnostics, 10(10), 854.
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2

Embryonic Protein Profile Analysis

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Total proteins extracted from embryos were analysed on a 10% SDS-PAGE followed by western blot analysis according to the procedures described by Lin et al. [5 (link)], except that the yolk was removed and the antibodies against Rtn4a (Abk; 1 : 1000), FAK (Cell Signaling; 1 : 1000), phosphor FAK [pY397] (Thermo; 1 : 1000), cell division control protein 42 homologue (cdc42) (New East; 1 : 500), active cdc42 (New East), α-tubulin (Sigma-Aldrich; 1 : 5000), GADPH (Santa Cruz; 1 : 1000), mouse-HRP (Santa Cruz; 1 : 5000) and rabbit-HRP (Santa Cruz; 1 : 5000) were used.
Lin C.Y., He J.Y., Zeng C.W., Loo M.R., Chang W.Y., Zhang P.H, & Tsai H.J. (2017). microRNA-206 modulates an Rtn4a/Cxcr4a/Thbs3a axis in newly forming somites to maintain and stabilize the somite boundary formation of zebrafish embryos. Open Biology, 7(7), 170009.
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3

Western Blot Analysis of YAP Protein

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To characterize Yap expression at the protein level, cells were lysed with ice-cold RIPA buffer and cleared by centrifuging at maximum speed for 5 minutes at 4 ºC. The total protein concentration in each sample was determined using the Pierce BCA assay kit. Equal protein amounts from each sample or treatment were then prepared and reduced in Laemmli sample buffer. The reduced samples were loaded and run in 10% Novex mini-cell SDS-PAGE gels (Invitrogen) and transferred to PVDF membranes (Bio Rad) after separation. The blots were then blocked in 5% BSA in TBS-T (blocking buffer) overnight at 4 ºC. Primary antibodies in blocking buffer were applied to the blots at room temperature for 2 hours. Blots were then rinsed with TBS-T, and secondary antibodies in TBS-T were applied for 2 hours. Chemiluminescent detection was used to visualize the blots, which were subsequently detected via autoradiography film (Hyblot CL). The film was processed and imaged using a Fotodyne white light transilluminator. Primary antibodies used were anti-YAP (H-9) (Santa Cruz Biotech.) and anti-β-actin (Santa Cruz Biotech.) at a dilution of 1:100 and 1:2000 respectively. The secondary antibody mouse-HRP (Santa Cruz Biotech.) was used at a 1:10,000 dilution. Densitometry analysis of the Western blots was carried out with ImageJ.
Kong Y.P., Rioja A.Y., Xue X., Sun Y., Fu J, & Putnam A.J. (2018). A Systems Mechanobiology Model to Predict Cardiac Reprogramming Outcomes on Different Biomaterials. Biomaterials, 181, 280-292.
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4

Protein Extraction and Western Blotting Methodology

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Cells were washed 3× with PBS, followed by lysis with MAPK buffer and a protease inhibitor cocktail from Sigma-Aldrich #P8340, 50 mM β-glycerophosphate, pH 7.2, 0.5% Triton X-100, 5 mM EGTA, 100 μM sodium orthovanadate, 1 mM dithiothreitol, and 2 mM MgCl2. 10–40 μg of total protein was fractionated by SDS–polyacrylamide gel electrophoresis and transferred to Polyvinyledine difuloride membranes. Antibodies used were as follows: pSTAT1 (Y701) Cell Signaling #9167S (1:500–1:1,000); STAT1, Cell Signaling #9172S (1:1,000–1:1,500); SOCS1, Abcam #ab3691 (1:300–1:500); IFNGR1 (interferon γ receptor α), Lifespan Biosciences #LS-C33-4260 (1:300–1:500); IFNGR2 (interferon γ receptor β/AF-1), Abcam #77246 (1:300–1:500); β-ACTIN, Sigma-Aldrich #A5441 (1: 5,000–1:10,000); Rabbit HRP, Jackson Immuno Research #111-035-144 (1: 5,000–1:10,000); and Mouse HRP, Santa Cruz #sc-2005 (1: 5,000–1:10,000).
Bullock B.L., Kimball A.K., Poczobutt J.M., Neuwelt A.J., Li H.Y., Johnson A.M., Kwak J.W., Kleczko E.K., Kaspar R.E., Wagner E.K., Hopp K., Schenk E.L., Weiser-Evans M.C., Clambey E.T, & Nemenoff R.A. (2019). Tumor-intrinsic response to IFNγ shapes the tumor microenvironment and anti–PD-1 response in NSCLC. Life Science Alliance, 2(3), e201900328.
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5

Protein Immunoblotting Protocol

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For immunoblotting, 20–40 μg of protein extracts were boiled and separated by 11–13% SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were transferred to polyvinylidene difluoride membranes, blocked for 1 h in Tris-buffered saline (TBS) containing 0.1% Tween 20 and 5% (w/v) dry skim milk powder, and incubated overnight with the indicated primary antibodies. The membranes were then incubated for 1 h with an HRP-conjugated secondary antibody (rabbit-HRP, Abcam; or mouse-HRP, Santa Cruz Biotechnology). Chemiluminescence was detected using chemiluminescence reagents (Animal Genetics, Tallahassee, FL, USA).
Park E.K., Lee J.C., Park J.W., Bang S.Y., Yi S.A., Kim B.K., Park J.H., Kwon S.H., You J.S., Nam S.W., Cho E.J, & Han J.W. (2015). Transcriptional repression of cancer stem cell marker CD133 by tumor suppressor p53. Cell Death & Disease, 6(11), e1964-.
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6

Glioma Cell Line Characterization

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The human glioma U-251 cell line was kindly provided by Dr. Dah-Yu Lu (Department of Pharmacology, School of Medicine, China Medical University, Taichung, Taiwan). The human glioma GBM8401 and GBM8901 cell line was kindly provided by Dr. Li-Sung Hsu (Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan). The U-87MG (BCRC No. 60360) and M059K cell line (BCRC No. 60381) were purchased from the Bioresources Collection and Research Center (BCRC, Hsinchu, Taiwan). These cells were cultured in DMEM/F12 and MEM medium supplemented with 1 × penicillin/streptomycin and 10% fetal bovine serum (HyClone, Logan, UT, USA) were incubated at 37 °C in a humidified incubator containing a 5% CO2 atmosphere. LCN2 antibody was purchased from R&D Systems, Inc (Minneapolis, MN, USA). U0126 (MEK inhibitor), siRNA-CTSD (si-CTSD), Cathepsin D (CTSD), t-ERK, β-actin, HRP-mouse and HRP-rabbit antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Matrigel Matrix was purchased from Corning company (Tewksbury, MA, USA). Phosphorylated-ERK (p-ERK) was obtained from Cell Signaling Technology (Beverly, MA, USA).
Hsieh Y.H., Tsai J.P., Yu C.L., Lee C.C., Hsu J.C, & Chen J.C. (2021). Overexpression of Lipocalin-2 Inhibits Proliferation and Invasiveness of Human Glioblastoma Multiforme Cells by Activating ERK Targeting Cathepsin D Expression. Biology, 10(5), 390.
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7

DNA Damage Response Protein Analysis

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Primary antibodies: RAD51 (Merk, Ab-1, PC130, 1:1000), RAD52 (Thermo Fisher, PA5-65036, 1:500), XAB2 (abcam, ab129487, 1:1000), CtIP (Active Motif, 61141, 1:500), ATM (abcam, ab32420 (Y170), 1:1000), P-ATM (abcam, ab81292 (pS1981), 1:50000), P-KAP1 (Bethyl, IHC-00073 (pS824) 1:200).
Secondary antibodies: HRP rabbit (Jackson Laboratory, Cat. No. 111-035-003, 1:50 000), HRP mouse (Amersham/Sigma, Cat. No. GENA931-1ML, 1:10 000), HRP mouse (Santa Cruz, sc-516102, 1:10 000).
Sharma A.B., Erasimus H., Pinto L., Caron M.C., Gopaul D., Peterlini T., Neumann K., Nazarov P.V., Fritah S., Klink B., Herold-Mende C.C., Niclou S.P., Pasero P., Calsou P., Masson J.Y., Britton S, & Van Dyck E. (2021). XAB2 promotes Ku eviction from single-ended DNA double-strand breaks independently of the ATM kinase. Nucleic Acids Research, 49(17), 9906-9925.
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8

MTT Assay for Apoptosis Evaluation

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3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) was obtained from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies against cleaved caspase-3, and cleaved PARP were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Mcl-1, β-actin, HRP-mouse and HRP-rabbit antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Hsieh Y.H., Yu F.J., Nassef Y., Liu C.J., Chen Y.S., Lin C.Y., Feng J.L, & Wu M.H. (2022). Targeting of Mcl-1 Expression by MiRNA-3614-5p Promotes Cell Apoptosis of Human Prostate Cancer Cells. International Journal of Molecular Sciences, 23(8), 4194.
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