Hrp conjugated β actin

Manufactured by Santa Cruz Biotechnology

Sourced in United States

HRP-conjugated β-actin is a laboratory reagent that consists of the protein β-actin conjugated to horseradish peroxidase (HRP). β-actin is a common structural protein found in eukaryotic cells, and the HRP conjugation allows for the detection and quantification of β-actin in biological samples using various immunoassay techniques.

Automatically generated - may contain errors

Lab products found in correlation

27 protocols using hrp conjugated β actin

Western Blot Analysis of Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates were processed and run in SDS/PAGE gels as described previously (Wang et al., 2014, 2016). Antibodies used to detect α‐tubulin, NANOG/NANOGP8 (sc‐134218), and HRP‐conjugated β‐actin were from Santa Cruz Biotechnology (Santa Cruz, CA). All other antibodies were from Cell Signaling (Beverly, MA, USA). Sizes of bands were estimated based on protein standards (Bio‐Rad, Hercules, CA, USA) on the membrane.

Wang R., Bhattacharya R., Ye X., Fan F., Boulbes D.R., Xia L, & Ellis L.M. (2017). Endothelial cells activate the cancer stem cell‐associated NANOGP8 pathway in colorectal cancer cells in a paracrine fashion. Molecular Oncology, 11(8), 1023-1034.

+ Open protocol

+ Expand

Silibinin Regulates Hedgehog Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

ASZ001 cells were received as a generous gift from Dr. Ervin Epstein (Children’s Hospital & Research Center Oakland) and cultured in M154F (Gibco, USA) media supplemented with 2% chelexed heat-inactivated FBS, 1% penicillin - streptomycin, and 0.05 M calcium chloride. Silibinin, dimethyl sulfoxide (DMSO) and trypan blue dye were purchased from Sigma-Aldrich (St. Louis, MO). Sant-1 was obtained from Tocris Biosciences (Bristol, UK); GDC-0449 was purchased from LC laboratories (Woburn, MA). The antibodies for phosphorylated-EGFR (Tyrosine-1173), EGFR, phosphorylated-Akt (Serine-473), phosphorylated-ERK1/2 (Threonine 202/Tyrosine 204), ERK1/2, cyclin D1 and HRP-conjugated secondary antibodies were purchased from Cell Signaling technology (Beverly, CA). Antibodies for Gli-1, SMO and SUFU were purchased from Abcam (Cambridge, UK). HRP-conjugated β-actin was purchased from Santa Cruz Biotechnology (CA, USA).

Dheeraj A., Rigby C.M., O’Bryant C.L., Agarwal C., Singh R.P., Deep G, & Agarwal R. (2017). Silibinin Treatment Inhibits the Growth of Hedgehog Inhibitor Resistant Basal Cell Carcinoma Cells via Targeting EGFR-MAPK-Akt and Hedgehog Signaling. Photochemistry and photobiology, 93(4), 999-1007.

+ Open protocol

+ Expand

Western Blot Analysis of Apoptosis Markers

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Gel electrophoresis and transfer was performed as described previously 18 (link). Using a Chemidoc XRS+ system (1708265; BIO-RAD; Hercules, CA, USA), proteins were detected with SuperSignal™ West Femto chemiluminescent solution (34095; Thermo Fisher Scientific, Waltham, MA, USA). Primary antibodies used in this study included the cleaved caspase 9 antibody (52873; 1:500 dilution; Cell signaling technology, Danvers, MA, USA), the cleaved caspase 3 antibody (9664; 1:500 dilution; Cell signaling technology), p16INK4A antibody (sc-1661; 1:500 dilution; Santa Cruz Biotechnology, Dallas, TX, USA), p21Cip1 antibody (sc-6246; 1:500 dilution; Santa Cruz), phospo-ERK antibody (9101s; 1:500 dilution; Cell signaling technology, Danvers, MA, USA) and HRP-conjugated β-actin (sc47778; 1:1000 dilution; Santa Cruz). Secondary antibodies included HRP-conjugated anti-mouse antibody (sc-516102; 1:1000 dilution; Santa Cruz) and HRP-conjugated anti-rabbit antibody (sc-2357; 1:1000dilution; Santa Cruz).

Lee Y.H., Kim M., Park H.J., Park J.Y., Song E.S., Lee H., Ko G., Ahn S., Kwon H.W., Byun Y., Kim C., Choi J, & Park J.T. (2023). Chemical screening identifies the anticancer properties of Polyporous parvovarius. Journal of Cancer, 14(1), 50-60.

+ Open protocol

+ Expand

Western Blotting for p53 and Lgr5

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blotting was performed with antibodies against p53 (FL‐393) and Lgr5 (Anti‐GPCR GPR49 antibody) purchased from Santa Cruz Biotechnology and Abcam, respectively. HRP‐conjugated β‐actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used as an internal control. Quantitative analysis was performed using the Image‐J.

Nakaoka T., Saito Y., Shimamoto Y., Muramatsu T., Kimura M., Kanai Y, & Saito H. (2017). Cluster microRNAs miR‐194 and miR‐215 suppress the tumorigenicity of intestinal tumor organoids. Cancer Science, 108(4), 678-684.

+ Open protocol

+ Expand

Western Blot Analysis of Protein Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates were processed and run in SDS-PAGE gels as described previously (Wang et al., 2014 (link); Wang et al., 2016 (link)). Antibodies used to detect α-tubulin, NANOG/NANOGP8 (sc-134218), and HRP conjugated β-actin were from Santa Cruz Biotechnology (Santa Cruz, CA). All other antibodies were from Cell Signaling (Beverly, MA). Sizes of bands were estimated based on protein standards (Bio Rad, Hercules, CA) on the membrane.

+ Open protocol

+ Expand

Evaluation of Protein Expression Levels

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot analysis was performed as described previously 18 (link). Primary antibodies used in this study p21Cip1 antibody (sc-6246; 1:500 dilution; Santa Cruz Biotechnology, Dallas, TX, USA), phospho-retinoblastoma (Rb) antibody (sc-271930; 1:500 dilution; Santa Cruz Biotechnology), phospo-ERK antibody (9101s; 1:500 dilution; Cell signaling technology, Danvers, MA, USA), p-MEK antibody (9121; 1:500 dilution; Cell signaling technology), Cleaved-Caspase 8 (9746s; 1:500 dilution; Cell signaling technology), Cleaved-Caspase 9 (9502S; 1:500 dilution; Cell signaling technology) and HRP-conjugated β-actin (sc47778; 1:1000 dilution; Santa Cruz Biotechnology). Secondary antibodies used in this study HRP conjugated anti-mouse antibody (sc-516102; 1:1000 dilution; Santa Cruz Biotechnology) and HRP conjugated anti-rabbit antibody (sc-2357; 1:1000 dilution; Santa Cruz Biotechnology).

Song E.S., So M.K., Park H.J., Lee H., Lee Y.H., Kuk M.U., Park J., Kwon H.W., Choi J, & Park J.T. (2023). Chemical screening identifies the anticancer properties of Polyporous tuberaster. Journal of Cancer, 14(11), 2075-2084.

+ Open protocol

+ Expand

Erlotinib and Cilengitide Modulate EMT Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Erlotinib was purchased from Sigma–Aldrich (St. Louis, MO, USA), and cilengitide was synthesized by Kyeong-Yong Park (CHA Meditech Co., Ltd., Daejeon, Korea) (Figures S1–S3). Fetal bovine serum (FBS), RPMI-1640 medium, and antibiotics (100 U/mL penicillin and 100 μg/mL streptomycin) were purchased from Corning Inc. (Corning, NY, USA). Recombinant human TGF-β1 was purchased from R&D Systems, Inc. (Minneapolis, MN, USA). Antibodies against E-cadherin (No. 3195), α-smooth muscle actin (α-SMA) (No. 19245), p-ERK1/2 (No. 4370), ERK1/2 (No. 4695), p-EGFR (No. 3777), EGFR (No. 4267), and p-Smad2/3 (No. 8828) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Antibodies against Smad2/3 (No. sc-133098), vimentin (No. sc-6260), N-cadherin (No. sc-59987), and β-catenin (No. sc-7199), horseradish peroxidase (HRP)-conjugated secondary antibodies (No. sc-516102 and No. sc-2357), HRP-conjugated actin (No. sc-1615), and HRP-conjugated β-actin (No. sc-47778) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Jeong J, & Kim J. (2022). Combination Effect of Cilengitide with Erlotinib on TGF-β1-Induced Epithelial-to-Mesenchymal Transition in Human Non-Small Cell Lung Cancer Cells. International Journal of Molecular Sciences, 23(7), 3423.

+ Open protocol

+ Expand

SDS-PAGE and Western Blot Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates were processed and run by SDS-PAGE gel electrophoresis as described previously (12 (link),13 (link)). Antibodies used to detect α-tubulin, and HRP conjugated β-actin were from Santa Cruz Biotechnology (Santa Cruz, CA). All other antibodies were from Cell Signaling (Beverly, MA). For each experiment, protein lysates were loaded to two gels and processed at the same time for separately probing for antibodies specific to phosphorylated proteins and total proteins. All membranes were probed with α-tubulin or β-actin as loading controls and a representative image was shown for each experiment. Each Western blot figure shows representative results of at least three independent experiments.

Wang R., Bhattacharya R., Ye X., Fan F., Boulbes D.R, & Ellis L.M. (2018). Endothelial Cells Promote Colorectal Cancer Cell Survival by Activating the HER3-AKT Pathway in a Paracrine Fashion. Molecular cancer research : MCR, 17(1), 20-29.

+ Open protocol

+ Expand

Molecular Signaling Pathways in Cancer Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

DS was purchased from Sigma-Aldrich (St. Louis, MO, USA). The compound was dissolved in 100% dimethyl sulfoxide (DMSO). A 50 mmol/L stock solution of DS was prepared and stored as small aliquots at –20°C until used. We purchased MTT, DMSO, gelatin, and horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit antibodies from Sigma-Aldrich. Fluorescein isothiocyanate (FITC)-conjugated CD44 and phycoerythrin (PE)-conjugated CD24 antibodies were purchased from BD Biosciences (San Jose, CA, USA). The phospho-specific antibodies anti-p38, anti-AKT, anti-mTOR, and specific antibodies anti-p38, anti-AKT, anti-mTOR, and AKT inhibitor LY294002 were purchased from Cell Signaling Technology (Danvers, MA, USA). HRP-conjugated β-actin, OCT4, proliferation cell nuclear antigen (PCNA), p53, p21, cdc2, cyclin B1, CDK4, cyclin D, CDK2, and cyclin E antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Ock C.W, & Kim G.D. (2021). Dioscin Decreases Breast Cancer Stem-like Cell Proliferation via Cell Cycle Arrest by Modulating p38 Mitogen-activated Protein Kinase and AKT/mTOR Signaling Pathways. Journal of Cancer Prevention, 26(3), 183-194.

+ Open protocol

+ Expand

VEGFR2 and STAT3 Phosphorylation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in RIPA buffer containing protease inhibitor cocktail (Roche Diagnostics, Indianapolis, IN) and orthovanadate (Thermo Scientifics, Rockford, IL), and clarified by centrifugation at 13,000×g rpm for 5 min at 4 °C. Protein concentration in the supernatant was quantified by bicinchoninic acid assay (BCA). 20 µg of protein from each treatment was loaded into NuPAGE 4–12% Bis-Tris Gels (Invitrogen, Carlsbad, CA), transferred to a PVDF membrane, and incubated with antibodies to phosphorylated VEGFR2 (p-VEGFR2, Y996) (1:500, Santa Cruz Biotechnology, Santa Cruz, CA), VEGFR2 (1:1000, Cell Signaling Technology, Danvers, MA), p-STAT3 (Y705) (1:1000, Cell Signaling Technology), or total STAT3 (1:1000, Cell Signaling Technology) at 4 °C overnight. After incubating with primary antibodies, membranes were then probed with HRP conjugated goat anti-rabbit secondary antibody or goat anti-mouse secondary antibody (1:3000–5000, ThermoFisher Scientific, Waltham, MA) at room temperature for 1 h. All the membranes were reprobed with HRP conjugated β-actin (1:3000, Santa Cruz Biotechnology) as loading controls. Densitometry analysis was performed on exposed films using UN-SCAN-IT software (Silk Scientific, Orem, UT) and normalized to β-actin.

Simmons A.B., Bretz C.A., Wang H., Kunz E., Hajj K., Kennedy C., Yang Z., Suwanmanee T., Kafri T, & Hartnett M.E. (2018). Gene therapy knockdown of VEGFR2 in retinal endothelial cells to treat retinopathy. Angiogenesis, 21(4), 751-764.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!