His Tag Antibody
His Tag Antibody Summary
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.
Data Examples
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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 human MYCOC and CHO Chinese hamster ovary cell line transfected with His-tagged human BAI-1. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-His Tag Monoclonal Antibody (Catalog # MAB050) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for His Tag at approximately 32, 65, and 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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Detection of His Tag in HEK293 Human Cell Line Transfected with His-tagged Proteins by Flow Cytometry.
HEK293 human embryonic kidney cell line transfected with His-tagged proteins was stained with Mouse Anti-His Tag Monoclonal Antibody (Catalog # MAB050, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer
(Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).
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Detection of His Tag by Simple WesternTM.
Simple Western lane view shows lysates of CHO Chinese hamster ovary cell line transfected with His-tagged human S1P4/EDG-6, loaded at 0.5 mg/mL. A specific band was detected for His-tagged S1P4/EDG-6 at approximately 75 kDa (as indicated) using 10 µg/mL of Mouse Anti-His Tag Monoclonal Antibody
(Catalog # MAB050). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
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 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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Particulate Array of Well-Ordered HIV Clade C Env Trimers Elicits Neutralizing Antibodies that Display a Unique V2 Cap Approach
Authors: P Martinez-M, K Tran, J Guenaga, G Lindgren, M Àdori, Y Feng, GE Phad, N Vázquez Be, S Bale, J Ingale, V Dubrovskay, S O'Dell, L Pramanik, M Spångberg, M Corcoran, K Loré, JR Mascola, RT Wyatt, GB Karlsson H
Immunity, 2017;46(5):804-817.e7.
Species: Primate
Sample Types: Plasma
Applications: ELISA Capture -
Biophysical analysis of the dynamics of calmodulin interactions with neurogranin and Ca2+/calmodulin-dependent kinase II
Authors: C Seeger, VO Talibov, UH Danielson
J. Mol. Recognit., 2017;0(0):.
Applications: Bioassay -
Phage display and selection of lanthipeptides on the carboxy-terminus of the gene-3 minor coat protein
Authors: JH Urban, MA Moosmeier, T Aumüller, M Thein, T Bosma, R Rink, K Groth, M Zulley, K Siegers, K Tissot, GN Moll, J Prassler
Nat Commun, 2017;8(1):1500.
Applications: Bioassay -
Structural basis of homo- and heterotrimerization of collagen I
Authors: U Sharma, L Carrique, S Vadon-Le G, N Mariano, RN Georges, F Delolme, P Koivunen, J Myllyharju, C Moali, N Aghajari, DJ Hulmes
Nat Commun, 2017;8(0):14671.
Species: Human
Sample Types: Protein
Applications: WB -
Discovery and Characterization of a Novel CD4-Binding Adnectin with Potent Anti-HIV Activity
Authors: D Wensel, Y Sun, Z Li, S Zhang, C Picarillo, T McDonagh, D Fabrizio, M Cockett, M Krystal, J Davis
Antimicrob. Agents Chemother., 2017;0(0):.
Applications: Binding Assay -
Hypoxia favors the generation of human plasma cells
Authors: M Schoenhals, M Jourdan, A Bruyer, A Kassambara, B Klein, J Moreaux
Cell Cycle, 2017;0(0):1-14.
Species: Human
Sample Types: Whole Cells
Applications: Functional Assay -
A Novel Bispecific Antibody Targeting EGFR and cMet that is Effective Against EGFR Inhibitor- Resistant Lung Tumors
Cancer Res, 2016;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Functional Assay -
Improving the developability of an anti-EphA2 single-chain variable fragment for nanoparticle targeting
Authors: Alexey A Lugovskoy
MAbs, 2016;0(0):0.
Applications: Flow -
I-bodies: human single domain antibodies that antagonize chemokine receptor CXCR4
Authors: K Griffiths, O Dolezal, B Cao, SK Nilsson, HB See, KD Pfleger, M Roche, PR Gorry, A Pow, K Viduka, K Lim, BG Lu, DH Chang, TA Murray-Rus, M Kvansakul, MA Perugini, C Dogovski, M Doerflinge, Y Zhang, K Parisi, JL Casey, SD Nuttall, M Foley
J Biol Chem, 2016;0(0):.
Species: Human
Sample Types: Complex Sample Type
Applications: Bioassay -
Thermostability of Well-Ordered HIV Spikes Correlates with the Elicitation of Autologous Tier 2 Neutralizing Antibodies
PLoS Pathog, 2016;12(8):e1005767.
Species: Guinea Pig
Sample Types: Serum
Applications: ELISA Capture -
HNRNPR Regulates the Expression of Classical and Nonclassical MHC Class I Proteins
J Immunol, 2016;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: WB -
Endotoxemia contributes to CD27+ memory B-cell apoptosis via enhanced sensitivity to Fas ligation in patients with Cirrhosis
Sci Rep, 2016;6(0):36862.
Species: Human
Sample Types: Whole Cells
Applications: Cross-linking -
Memo has a novel role in S1P signaling and is [corrected] crucial for vascular development.
Authors: Kondo, Shunya, Bottos, Alessia, Allegood, Jeremy C, Masson, Regis, Maurer, Francisc, Genoud, Christel, Kaeser, Patrick, Huwiler, Andrea, Murakami, Masato, Spiegel, Sarah, Hynes, Nancy E
PLoS ONE, 2014;9(4):e94114.
Species: Human
Sample Types: Cell Lysates
Applications: WB -
VB-201, an oxidized phospholipid small molecule, inhibits CD14- and Toll-like receptor-2-dependent innate cell activation and constrains atherosclerosis.
Authors: Mendel I, Feige E, Yacov N, Salem Y, Levi I, Propheta-Meiran O, Shoham A, Ishai E, George J, Harats D, Breitbart E
Clin Exp Immunol, 2014;175(1):126-37.
Species: Human
Sample Types: Whole Cells
Applications: Flow -
Generation of a high-fidelity antibody against nerve growth factor using library scanning mutagenesis and validation with structures of the initial and optimized Fab-antigen complexes.
MAbs, 2014;6(4):1059-68.
Species: N/A
Sample Types: Chip
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Galectin-9 controls the therapeutic activity of 4-1BB-targeting antibodies.
Authors: Madireddi S, Eun S, Lee S, Nemcovicova I, Mehta A, Zajonc D, Nishi N, Niki T, Hirashima M, Croft M
J Exp Med, 2014;211(7):1433-48.
Species: Mouse
Sample Types: Whole Cells
Applications: Cross-linking -
Novel septin 9 repeat motifs altered in neuralgic amyotrophy bind and bundle microtubules.
Authors: Bai X, Bowen J, Knox T, Zhou K, Pendziwiat M, Kuhlenbaumer G, Sindelar C, Spiliotis E
J Cell Biol, 2013;203(6):895-905.
Species: Bovine
Sample Types: Recombinant Protein
Applications: WB -
Cross-species withdrawal of MCL1 facilitates postpartum uterine involution in both the mouse and baboon.
Authors: Kyathanahalli, Chandras, Marks, Jason, Nye, Kennedy, Lao, Belinda, Albrecht, Eugene D, Aberdeen, Graham W, Nathanielsz, Peter W, Jeyasuria, Panchara, Condon, Jennifer
Endocrinology, 2013;154(12):4873-84.
Species: Human
Sample Types: Whole Cells
Applications: Cross-linking -
Pigment epithelium-derived factor (PEDF) binds to caveolin-1 and inhibits the pro-inflammatory effects of caveolin-1 in endothelial cells.
Authors: Matsui T, Higashimoto Y, Taira J, Yamagishi S
Biochem Biophys Res Commun, 2013;441(2):405-10.
Species: Human
Sample Types: Cell Lysates
Applications: WB -
Mutation of Y407 in the CH3 domain dramatically alters glycosylation and structure of human IgG.
Authors: Rose, Rebecca, van Berkel, Patrick, van den Bremer, Ewald T, Labrijn, Aran F, Vink, Tom, Schuurman, Janine, Heck, Albert J, Parren, Paul W H
MAbs, 2013;5(2):219-28.
Species: Human
Sample Types: Recombinant Protein
Applications: ELISA Development -
Antiapoptotic signaling via MCL1 confers resistance to caspase-3-mediated apoptotic cell death in the pregnant human uterine myocyte.
Authors: Stephenson-Famy A, Marks J, Suresh A
Mol. Endocrinol., 2012;26(2):320-30.
Species: N/A
Sample Types: Recombinant Protein
Applications: Cross-linking -
Comparison of epithelial differentiation and immune regulatory properties of mesenchymal stromal cells derived from human lung and bone marrow.
Authors: Ricciardi M, Malpeli G, Bifari F, Bassi G, Pacelli L, Nwabo Kamdje AH, Chilosi M, Krampera M
PLoS ONE, 2012;7(5):e35639.
Species: Human
Sample Types: Recombinant Protein
Applications: Cross-linking -
C17 prevents inflammatory arthritis and associated joint destruction in mice.
Authors: Chao C, Joyce-Shaikh B, Grein J
PLoS ONE, 2011;6(7):e22256.
Species: Mouse
Sample Types: Serum
Applications: ELISA Development -
A novel role for reciprocal CD30-CD30L signaling in the cross-talk between natural killer and dendritic cells.
Authors: Simhadri VL, Hansen HP, Simhadri VR, Reiners KS, Bessler M, Engert A, von Strandmann EP
Biol. Chem., 2011;393(1):101-6.
Species: N/A
Sample Types: Recombinant Protein
Applications: Cross-linking -
Active 1918 pandemic flu viral neuraminidase has distinct N-glycan profile and is resistant to trypsin digestion.
Authors: Wu ZL, Ethen C, Hickey GE, Jiang W
Biochem. Biophys. Res. Commun., 2009;379(3):749-53.
Species: Virus
Sample Types: Recombinant Protein
Applications: WB -
Targeting of bone morphogenetic protein growth factor complexes to fibrillin.
Authors: Sengle G, Charbonneau NL, Ono RN, Sasaki T, Alvarez J, Keene DR, Bachinger HP, Sakai LY
J. Biol. Chem., 2008;283(20):13874-88.
Species: N/A
Sample Types: Recombinant Protein
Applications: Immunopurification -
A new model for growth factor activation: type II receptors compete with the prodomain for BMP-7.
Authors: Sengle G, Ono RN, Lyons KM, Bachinger HP, Sakai LY
J. Mol. Biol., 2008;381(4):1025-39.
Species: Human
Sample Types: Recombinant Protein
Applications: ELISA Development -
The Fas/Fas ligand system inhibits differentiation of murine osteoblasts but has a limited role in osteoblast and osteoclast apoptosis.
Authors: Kovacic N, Lukic IK, Grcevic D, Katavic V, Croucher P, Marusic A
J. Immunol., 2007;178(6):3379-89.
Species: N/A
Sample Types: Recombinant Protein
Applications: Cross-linking -
Mannan-binding lectin-associated serine protease 3 cleaves synthetic peptides and insulin-like growth factor-binding protein 5.
Authors: Cortesio CL, Jiang W
Arch. Biochem. Biophys., 2006;449(1):164-70.
Species: N/A
Sample Types: Recombinant Protein
Applications: WB -
Rapid induction of apoptosis in B-cell lymphoma by functionally isolated human antibodies.
Authors: Fransson J, Tornberg UC, Borrebaeck CA, Carlsson R, Frendeus B
Int. J. Cancer, 2006;119(2):349-58.
Species: Human
Sample Types: Whole Cells
Applications: Flow -
Receptor-ligand binding in the cell-substrate contact zone: a quantitative analysis using CX3CR1 and CXCR1 chemokine receptors.
Authors: Lee FH, Haskell C, Charo IF, Boettiger D
Biochemistry, 2004;43(22):7179-86.
Species: Mouse
Sample Types: Whole Cells
Applications: Binding Assay -
Topological analysis of Hedgehog acyltransferase, a multipalmitoylated transmembrane protein.
Authors: Konitsiotis A, Jovanovic B, Ciepla P, Spitaler M, Lanyon-Hogg T, Tate E, Magee A
J Biol Chem, 0;290(6):3293-307.
Species: Human
Sample Types: Cell Lysates
Applications: WB
FAQs
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This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?
The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.
Isotype Controls
Primary Antibodies
Reconstitution Buffers
Secondary Antibodies
Reviews for His Tag Antibody
Average Rating: 4.8 (Based on 6 Reviews)
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293T cells were transfected with pCDNA 3.1-Control or His-RIP1 for 48h. Total cell lysates were subjected to western blot. PVDF membrane were probed with 1 um/ml Human His Tag Antibody (MAB050). A specific band was detected for His-RIP1 at approximately 75 kDa. This experiment was conducted under reducing conditions
cell lysates transformed
Specificity: Specific
Sensitivity: Sensitive
Buffer: TBS + Tween + BSA
Dilution: used at 2ug/mL
