His Tag Horseradish Peroxidase-conjugated Antibody Summary
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 μm filtered solution in PBS.
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
Detection of His Tag by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line either non-transfected or transfected with His-tagged human CIQ4 and CHO Chinese hamster ovary cell line transfected with His-tagged BAI-1. PVDF membrane was probed with a 1:4000 dilution of Mouse Anti-His Tag HRP-conjugated Monoclonal Antibody (Catalog # MAB050H). Specific bands were detected for His Tag at approximately 32 and 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of His Tag by Simple WesternTM. Simple Western lane view shows lysates of HEK293 human embryonic kidney cell line transfected with His-tagged human CIQ4 and human MYCOC and CHO Chinese hamster ovary cell line transfected with His-tagged BAI-1, loaded at 0.2 mg/mL. Specific bands were detected for His Tag at approximately 32, 65, and 120 kDa (as indicated) using a 1:1000 dilution of Mouse Anti-His Tag HRP-conjugated Monoclonal Antibody (Catalog # MAB050H). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Reconstitution Calculator
Preparation and Storage
- 6 months from date of receipt, 2 to 8 °C as supplied.
Background: His Tag
Consecutive histidine residues (usually 6 to 10 in length) are often inserted into the amino acid sequences of recombinant proteins. The resulting His-tagged proteins can be detected or purified by using anti-polyHis antibodies.
Product Datasheets
Citations for His Tag Horseradish Peroxidase-conjugated Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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A conserved C-terminal domain of TamB interacts with multiple BamA POTRA domains in Borreliella burgdorferi
Authors: Hall, KT;Kenedy, MR;Johnson, DK;Hefty, PS;Akins, DR;
PloS one
Species: E. coli
Sample Types: Cell Lysates
Applications: Western Blot -
Fibrin drives thromboinflammation and neuropathology in COVID-19
Authors: Ryu, JK;Yan, Z;Montano, M;Sozmen, EG;Dixit, K;Suryawanshi, RK;Matsui, Y;Helmy, E;Kaushal, P;Makanani, SK;Deerinck, TJ;Meyer-Franke, A;Rios Coronado, PE;Trevino, TN;Shin, MG;Tognatta, R;Liu, Y;Schuck, R;Le, L;Miyajima, H;Mendiola, AS;Arun, N;Guo, B;Taha, TY;Agrawal, A;MacDonald, E;Aries, O;Yan, A;Weaver, O;Petersen, MA;Meza Acevedo, R;Alzamora, MDPS;Thomas, R;Traglia, M;Kouznetsova, VL;Tsigelny, IF;Pico, AR;Red-Horse, K;Ellisman, MH;Krogan, NJ;Bouhaddou, M;Ott, M;Greene, WC;Akassoglou, K;
Nature
Species: N/A
Sample Types: Antibody
Applications: ELISA Detection -
Rapid discovery of monoclonal antibodies by microfluidics-enabled FACS of single pathogen-specific antibody-secreting cells
Authors: Fischer, K;Lulla, A;So, TY;Pereyra-Gerber, P;Raybould, MIJ;Kohler, TN;Yam-Puc, JC;Kaminski, TS;Hughes, R;Pyeatt, GL;Leiss-Maier, F;Brear, P;Matheson, NJ;Deane, CM;Hyvönen, M;Thaventhiran, JED;Hollfelder, F;
Nature biotechnology
Species: Human
Sample Types: Whole Cells
Applications: Neutralization -
Immuno-profiling of Brucella proteins for developing improved vaccines and DIVA capable serodiagnostic assays for brucellosis
Authors: Nandini, P;Jakka, P;Murugan, S;Mazumdar, V;Kumar, D;Prakash, R;Barbuddhe, SB;Radhakrishnan, G;
Frontiers in microbiology
Species: Bacteria
Sample Types: Cell Lysates
Applications: Western Blot -
Preclinical Characterization and Phase I Trial Results of INBRX-109, A Third-Generation, Recombinant, Humanized, Death Receptor 5 Agonist Antibody, in Chondrosarcoma
Authors: Vivek Subbiah, Sant P. Chawla, Anthony P. Conley, Breelyn A. Wilky, Anthony Tolcher, Nehal J. Lakhani et al.
Clinical Cancer Research
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Shifting the balance: soluble ADAM10 as a potential treatment for Alzheimer's disease
Authors: Ayelet Sarah Hershkovits, Sivan Gelley, Rawad Hanna, Oded Kleifeld, Avidor Shulman, Ayelet Fishman
Frontiers in Aging Neuroscience
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Systemic checkpoint blockade by PD-L1 single-chain antibody confers potent anti-tumor immunity and long-term survival
Authors: H Wang, V Khattar, JA Hensel, R Ashton, Y Lu, AG Sorace, Y Wang, JS Deshane, JL Mieher, C Deivanayag, S Ponnazhaga
Molecular Cancer Therapeutics, 2022-11-03;0(0):.
Species: Mouse
Sample Types: Cell Lysate
Applications: Western Blot -
PIM1-Induced Cytoplasmic Expression of RBMY Mediates Hepatocellular Carcinoma Metastasis
Authors: HH Chua, MH Chang, YH Chen, DJ Tsuei, YM Jeng, PH Lee, YH Ni
Cellular and Molecular Gastroenterology and Hepatology, 2022-10-01;0(0):.
Species: Human, Mouse
Sample Types: Tissue Homogenates
Applications: Western Blot -
Comprehensive fitness landscape of SARS-CoV-2 Mpro reveals insights into viral resistance mechanisms
Authors: Julia M Flynn, Neha Samant, Gily Schneider-Nachum, David T Barkan, Nese Kurt Yilmaz, Celia A Schiffer et al.
eLife
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Comparative expression of soluble, active human kinases in specialized bacterial strains
Authors: A Sunderhaus, R Imran, E Enoh, A Adedeji, T Obafemi, MH Abdel Aziz
PLoS ONE, 2022-04-19;17(4):e0267226.
Species: Bacteria - Escherichia coli
Sample Types: Cell Lysates
Applications: Western Blot -
Dendritic cell immunoreceptor drives atopic dermatitis by modulating oxidized CaMKII-involved mast cell activation
Authors: X Luo, J Chen, H Yang, X Hu, MP Alphonse, Y Shen, Y Kawakami, X Zhou, W Tu, T Kawakami, M Wan, NK Archer, H Wang, P Gao
JCI Insight, 2022-03-08;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: ELISA Detection -
Evaluation of Immunogenicity and Efficacy of Fasciola hepatica Tetraspanin 2 (TSP2) Fused to E. coli Heat-Labile Enterotoxin B Subunit LTB Adjuvant Following Intranasal Vaccination of Cattle
Authors: G Zerna, VA Rathinasam, H Toet, G Anderson, R Dempster, TW Spithill, T Beddoe
Vaccines, 2021-10-20;9(11):.
Species: Yeast
Sample Types: Cell Lysates
Applications: ELISA Development, Western Blot -
Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon alpha2b
Authors: X Qin, M Duan, D Pei, J Lin, L Wang, P Zhou, W Yao, Y Guo, X Li, L Tao, Y Ding, L Liu, Y Zhou, C Jia, C Rao, J Wang
Journal of pharmaceutical analysis, 2021-07-08;12(2):308-316.
Species: Human
Sample Types: N/A
Applications: ELISA Detection -
Structural basis for broad coronavirus neutralization
Authors: Maximilian M. Sauer, M. Alejandra Tortorici, Young-Jun Park, Alexandra C. Walls, Leah Homad, Oliver J. Acton et al.
Nature Structural & Molecular Biology
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Characterization of a profilin-like protein from Fasciola hepatica
Authors: J Wilkie, TC Cameron, T Beddoe
PeerJ, 2020-12-07;8(0):e10503.
Species: Fasciolosis
Sample Types: Recombinant Protein
Applications: Western Blot -
S-nitrosylation-mediated activation of a histidine kinase represses the type 3 secretion system and promotes virulence of an enteric pathogen
Authors: D Gu, Y Zhang, Q Wang, X Zhou
Nat Commun, 2020-11-13;11(1):5777.
Species: Vibrio parahaemolyticus
Sample Types: Cell Lysates
Applications: Western Blot -
Phosphatidylinositol-3-OH kinase signalling is spatially organized at endosomal compartments by microtubule-associated protein 4
Authors: N Thapa, M Chen, HT Horn, S Choi, T Wen, RA Anderson
Nat Cell Biol, 2020-11-02;22(11):1357-1370.
Species: Human
Sample Types: Protein
Applications: Functional Assay -
Histone Methyltransferase DOT1L is Involved in Larval Molting and Second Stage Nymphal Feeding in Ornithodoros Moubata
Authors: J Gobl, D Kumar Sinh, R Sima, J Perner, P Kopá?ek, JJ Valdés, ROM Rego, A Cabezas-Cr
Vaccines (Basel), 2020-04-01;8(2):.
Species: Human
Sample Types: Protein
Applications: Western Blot -
Sensitivity of Kupffer cells and liver sinusoidal endothelial cells to ricin toxin and ricin toxin-Ab complexes
Authors: B Mooney, FJ Torres-Vel, J Doering, DJ Ehrbar, NJ Mantis
J. Leukoc. Biol., 2019-07-16;0(0):.
Species: Human, Mouse
Sample Types: Protein
Applications: Binding Assay -
Dual regulation of Arabidopsis AGO2 by arginine methylation
Authors: P Hu, H Zhao, P Zhu, Y Xiao, W Miao, Y Wang, H Jin
Nat Commun, 2019-02-19;10(1):844.
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A Periplasmic Antimicrobial Peptide-Binding Protein Is Required for Stress Survival in Vibrio cholerae
Authors: J Saul-McBet, JS Matson
Front Microbiol, 2019-02-05;10(0):161.
Applications: Western Blot -
Functional genomic characterization of a synthetic anti-HER3 antibody reveals a role for ubiquitination by RNF41 in the anti-proliferative response
Authors: JP Turowec, EWT Lau, X Wang, KR Brown, FA Fellouse, KK Jawanda, J Pan, J Moffat, S Sidhu
J. Biol. Chem., 2018-12-06;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Spherical-supported membranes as platforms for screening against membrane protein targets
Authors: V. Vasilca, A. Sadeghpour, S. Rawson, L.E. Hawke, S.A. Baldwin, T. Wilkinson et al.
Analytical Biochemistry
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Structural basis for substrate selection by the translocation and assembly module of the beta-barrel assembly machinery
Authors: Rebecca S. Bamert, Karl Lundquist, Hyea Hwang, Chaille T. Webb, Takoya Shiota, Christopher J. Stubenrauch et al.
Molecular Microbiology
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CD36 Differently Regulates Macrophage Responses to Smooth and Rough Lipopolysaccharide
Authors: R Biedro?, A Peru?, S Józefowski
PLoS ONE, 2016-04-13;11(4):e0153558.
Species: Mouse
Sample Types: Recombinant Protein
Applications: Binding Assay -
Extracellular Regulation of Bone Morphogenetic Protein Activity by the Microfibril Component Fibrillin-1.
Authors: Wohl A, Troilo H, Collins R, Baldock C, Sengle G
J Biol Chem, 2016-04-08;291(24):12732-46.
Species: Human
Sample Types: Protein
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Oxidation by neutrophils-derived HOCl increases immunogenicity of proteins by converting them into ligands of several endocytic receptors involved in antigen uptake by dendritic cells and macrophages.
Authors: Biedron R, Konopinski M, Marcinkiewicz J, Jozefowski S
PLoS ONE, 2015-04-07;10(4):e0123293.
Species: Mouse
Sample Types: Protein
Applications: ELISA Development (Detection) -
A Nonfucosylated Variant of the anti-HIV-1 Monoclonal Antibody b12 Has Enhanced Fc gamma RIIIa-Mediated Antiviral Activity In Vitro but Does Not Improve Protection against Mucosal SHIV Challenge in Macaques
Authors: Brian Moldt, Mami Shibata-Koyama, Eva G. Rakasz, Niccole Schultz, Yutaka Kanda, D. Cameron Dunlop et al.
Journal of Virology
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Thiol isomerases negatively regulate the cellular shedding activity of ADAM17
Authors: Sofie H. Willems, Christopher J. Tape, Peter L. Stanley, Neil A. Taylor, Ian G. Mills, David E. Neal et al.
Biochemical Journal
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Antagonists to human and mouse vascular endothelial growth factor receptor 2 generated by directed protein evolution in vitro.
Authors: Getmanova EV, Chen Y, Bloom L, Gokemeijer J, Shamah S, Warikoo V, Wang J, Ling V, Sun L
Chem. Biol., 2006-05-01;13(5):549-56.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry
FAQs
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Can the His Tag Antibody (MAB050) recognize all His-tagged proteins?
MAB050 has been validated specifically for the detection of His-tags localized to the amino- or carboxyl-terminus of a peptide. It is also expected to detect any series of 6 or more histidine residues as long as this epitope is conformationally accessible. For applications utilizing linearized proteins, it is likely that MAB050 will have access to any consecutive histidine regions. If used with protein in its native conformation, there is the potential that the protein will be folded such that the antibody is not able to bind its target.
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What is the molecular weight of an HRP-conjugated antibody?
The average ratio of HRP:antibody is estimated to be 2 to 4 HRP molecules per antibody molecule based on A403 and A280 readings of the final conjugate. Using ~44kDa as the molecular weight of HRP and 150kDa as the molecular weight of IgG, the theoretical average MW of the conjugated antibody is 238kDa to 326kDa. The HRP-conjugation does vary quite a bit with each antibody, but does not vary as much with each conjugation of the same antibody, especially if it is a monoclonal.
Reviews for His Tag Horseradish Peroxidase-conjugated Antibody
Average Rating: 5 (Based on 6 Reviews)
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This antibody works very well to detect his-tagged antigens via Western blot. In the blot shown here, the His antibody was used to detect recombinant TcfA, TcfB, and TcfC. The bands are clear with little to no background. Because this is HRP labeled, there is no need for a secondary antibody, which is helpful to speed up the assay. I will use a secondary to increase band intensity.
Western blot was made in 5% BSA, antibody worked beautifully at 1:1000 dilution.
Excellent Antibody. Detects 6xhis positive control, almost instantaneously.