Hrp protein a

Manufactured by BD

Sourced in United States

HRP Protein A is a recombinant protein that binds to the Fc region of immunoglobulins. It is commonly used as a detection reagent in various immunoassay techniques, such as ELISA and Western blotting, to identify and quantify the presence of specific antibodies in samples.

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Lab products found in correlation

Protease inhibitor cocktail, by Merck Group (2 mentions) Horseradish peroxidase hrp conjugated secondary antibody, by Merck Group (2 mentions) Rabbit anti ap4 ε, by BD (2 mentions) Vinculin, by Merck Group (1 mentions) Protein g dynabead, by Thermo Fisher Scientific (1 mentions) Cenpf, by Abcam (1 mentions) Top2a, by Santa Cruz Biotechnology (1 mentions) Cdc20, by Santa Cruz Biotechnology (1 mentions) Flag antibody, by Merck Group (1 mentions) Anti flag m2, by Merck Group (1 mentions) Anti ha 11, by Fortrea (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Anti β actin, by Merck Group (1 mentions) Chloral hydrate, by Merck Group (1 mentions) Goat anti mouse igg hrp, by Santa Cruz Biotechnology (1 mentions) Fam labeled taqman primers, by Thermo Fisher Scientific (1 mentions) Surebeads protein g magnetic beads, by Bio-Rad (1 mentions) Any kd mini protean tgx gel, by Bio-Rad (1 mentions) Gene expression master mix, by Thermo Fisher Scientific (1 mentions) Nitrocellulose filter paper sandwiches, by Bio-Rad (1 mentions)

4 protocols using hrp protein a

Western Blot and Immunoprecipitation Assay

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Whole protein extracts were obtained by lysing cells in TNN buffer (50mM Tris (pH 7.5), 120mM NaCl, 5mM EDTA, 0.5% NP40, 10mM Na4P₂O₇, 2mM Na₃VO₄, 100mM NaF, 1mM PMSF, 1mM DTT, 10mM β-glycerophosphate, protease inhibitor cocktail (Sigma)). Protein lysates or purified protein were separated by SDS-PAGE, transferred to a PVDF-membrane and detected by immunoblotting with the first and secondary antibodies: β-actin (Santa Cruz Biotechnology, sc-47778) 1:5000, B-MYB (clone LX015.1, [42 (link)] 1:5, anti-HA.11 (Covance, MMA-101P) 1:1000, anti-FLAG M2 (Sigma, F3165) 1:5000, anti-His (Sigma, H1029) 1:2000, Vinculin (Sigma, V9131) 1:10000, TOP2A (Santa Cruz Biotechnology, sc-365916) 1:1000, CDC20 (Santa Cruz Biotechnology, sc-5296) 1:500, YAP (Santa Cruz Biotechnology, sc-10199) 1:1000, p-YAP(S127A) (Cell Signaling Technology, 4911) 1:1000, LIN9 (Bethyl, A300-BL2981), NUSAP1 (Geert Carmeliet) 1:1000, CENPF (Abcam, ab-5) 1:1000, anti-mouse-HRP (GE healthcare, NXA931) 1:5000 and HRP Protein A (BD Biosciences, 610438) 1:5000. For immunoprecipitation of FLAG-tagged proteins, protein G dynabeads (Thermo Fisher Scientific) were first coupled with 1 μg FLAG-antibody (Sigma, F3165) and then immunoprecipitated with 1mg whole cell lysate. After five times of washing with TNN, proteins were separated by SDS-PAGE and detected by immunoblotting using the desired antibodies.

Gründl M., Walz S., Hauf L., Schwab M., Werner K.M., Spahr S., Schulte C., Maric H.M., Ade C.P, & Gaubatz S. (2020). Interaction of YAP with the Myb-MuvB (MMB) complex defines a transcriptional program to promote the proliferation of cardiomyocytes. PLoS Genetics, 16(5), e1008818.

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Characterization of Cannabinoid Receptor Signaling

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Fluriso™ was purchased from VetOne (Boise, ID, USA). Chloral hydrate, sodium pentobarbital and protease inhibitor cocktail were purchased from Sigma-Aldrich (St. Louis, MO, USA). Lentiviral vectors were produced and packaged by VectorBuilder (Santa Clara, CA, USA). The Pierce ECL Western Blotting Substrate, Tris/glycine/SDS buffer RNAqueous®, 4PCR Total RNA Isolation Kit and High Capacity cDNA Reverse Transcriptase Kits, as well as FAM-labeled TaqMan Primers and Gene Expression Master Mix were purchased from ThermoFischer Scientific (Waltham, MA, USA). Anti-CB1R and anti-CNRIP1 antibodies were purchased from Abeam (Cambridge, MA, USA). Anti-CNR1 antibody (used in western blot) was purchased from Synaptic Systems (Goettingen, Germany). HRP Protein A was purchased from BD Biosciences (San Jose, CA, USA). Anti-β-actin was purchased from Chemicon. Goat anti-rabbit IgG-HRP and goat anti-mouse IgG-HRP were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). SureBeads™ Protein G Magnetic beads, Laemmli Sample Buffer, Mini-Protean TGX Any kD gels and nitrocellulose/filter paper sandwiches were supplied from BioRad (Hercules, CA, USA). All other chemicals and reagents were of either analytical or laboratory grade, and were purchased from various suppliers.

Perez S.M., Donegan J.J., Boley A.M., Aguilar D.D., Giuffrida A, & Lodge D.J. (2018). Ventral hippocampal overexpression of Cannabinoid Receptor Interacting Protein 1 (CNRIP1) produces a schizophrenia-like phenotype in the rat. Schizophrenia research, 206, 263-270.

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Antibody Characterization and Application

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Unless otherwise noted, all chemicals were purchased from Sigma (St.
Louis, MO, USA). The following antibodies were used in this study: rabbit
anti-AP4 β, rabbit anti-AP4 ε (for western blotting and
immunoprecipitation; both in-house; Hirst et al., 1999), mouse anti-AP4
ε (for immunofluorescence; 612019; BD Transduction Labs), rabbit
anti-clathrin (in-house; Simpson et al., 1996), rabbit anti-GFP (gift from
Matthew Seaman, Cambridge Institute for Medical Research, UK), and rabbit
anti-tepsin (in-house; Borner et al., 2012). Horseradish peroxidase
(HRP)–conjugated secondary antibodies were purchased from Sigma-Aldrich,
and fluorescently labelled secondary antibodies were from Invitrogen. For
Western blotting of immunoprecipitates where the protein band of interest was
close to an IgG band, protein-A-HRP (BD Biosciences) was used in the place of
HRP-conjugated secondary antibody.

Archuleta T.L., Frazier M.N., Monken A.E., Kendall A.K., Harp J., McCoy A.J., Creanza N, & Jackson L.P. (2017). Structure and evolution of ENTH and VHS/ENTH-like domains in tepsin. Traffic (Copenhagen, Denmark), 18(9), 590-603.

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Immunoprecipitation and Western Blotting

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Unless otherwise noted, all chemicals were purchased from Sigma (St. Louis, MO, USA). The following antibodies were used in this study: rabbit anti-AP4 β, rabbit anti-AP4 ε (for western blotting and immunoprecipitation; both in-house; Hirst et al., 1999), mouse anti-AP4 ε (for immunofluorescence; 612019; BD Transduction Labs), rabbit anti-clathrin (in-house; Simpson et al., 1996), rabbit anti-GFP (gift from Matthew Seaman, Cambridge Institute for Medical Research, UK), and rabbit anti-tepsin (in-house; Borner et al., 2012). Horseradish peroxidase (HRP)–conjugated secondary antibodies were purchased from Sigma-Aldrich, and fluorescently labelled secondary antibodies were from Invitrogen. For Western blotting of immunoprecipitates where the protein band of interest was close to an IgG band, protein-A-HRP (BD Biosciences) was used in the place of HRP-conjugated secondary antibody.

Frazier M.N., Davies A.K., Voehler M., Kendall A.K., Borner G.H., Chazin W.J., Robinson M.S, & Jackson L.P. (2016). Molecular basis for the interaction between Adaptor Protein Complex 4 (AP4) β4 and its accessory protein, tepsin. Traffic (Copenhagen, Denmark), 17(4), 400-415.

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