Histidine Tag antibody | AD1.1.10
Horseradish peroxidase conjugated Mouse anti Histidine tag antibody clone AD1.1.0 (MCA1396P) used for the detection of recombinant scTCR varients by western blotting.
Image caption:
4B2A1 scTCR and rFkpA expression level. Western blot showing periplasmic expression levels of 4B2A1 scTCR variants and rFkpA from pHOG, pFKPEN (pEN) and pFKPEI (pEI) vectors under different expression conditions, detected with anti-His tag -HRP Ab and anti-Flag- M2 HRP Ab as described. The scTCR and rFkpA were developed with SuperSignal West Pico substrate and exposed to BioMax MR film for for 30 min and 30 sec, respectively, for the upper and lower panels. The expression analysis was repeated twice, and representative results are shown.
From: Gunnarsen KS, Lunde E, Kristiansen PE, Bogen B, Sandlie I, Løset GÅ.
Periplasmic expression of soluble single chain T cell receptors is rescued by the chaperone FkpA.
BMC Biotechnol. 2010 Feb 3;10:8.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.
Horseradish peroxidase conjugated Mouse anti Histidine tag antibody clone AD1.1.0 (MCA1396P) used for the detection of recombinant scTCR varients by western blotting.
Image caption:
7A10B2 scTCR and rFkpA expression level. Western blot showing periplasmic expression levels of 7A10B2 scTCR variants and rFkpA from pHOG, pFKPEN (pEN) and pFKPEI (pEI) vectors under different expression conditions, detected with anti-His tag-HRP and anti-Flag-M2-HRP as described. The scTCR and rFkpA were developed with SuperSignal West Pico substrate and exposed to BioMax MR film for for 30 min and 30 sec, respectively, for the upper and lower panels. The expression analysis was repeated twice, and representative results are shown.
From: Gunnarsen KS, Lunde E, Kristiansen PE, Bogen B, Sandlie I, Løset GÅ.
Periplasmic expression of soluble single chain T cell receptors is rescued by the chaperone FkpA.
BMC Biotechnol. 2010 Feb 3;10:8.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.
Horseradish peroxidase conjugated Mouse anti Histidine tag antibody clone AD1.1.0 (MCA1396P) used for the detection of recombinant scTCR varients by western blotting.
Image caption:
Purity of proteins. (A) and (B); S75 size exclusion chromatography of scTCR 4B2A1 Vαβ and Vβα respectively. 50 μl of the IMAC purified material was subject to the purification procedure. In (A) fractions A14-B1 corresponds to the higher aggregates, while fraction B5-B7 corresponds to the monomeric scTCR 4B2A1 Vαβ, the monomeric fraction is highlighted with an arrow. In (B) fraction A15-B1 and B3-B4 corresponds to the higher aggregates, while fraction B6-B7 corresponds to the monomeric scTCR 4B2A1 Vβα, the monomeric fraction is highlighted with an arrow. (C) and (D); Western blots showing reduced and non-reduced scTCR Vαβ and Vβα after IMAC purification and S75 size exclusion chromatography. Samples of 0.25 μg were loaded on the gel and analysed with anti-His tag-HRP.
From: Gunnarsen KS, Lunde E, Kristiansen PE, Bogen B, Sandlie I, Løset GÅ.
Periplasmic expression of soluble single chain T cell receptors is rescued by the chaperone FkpA.
BMC Biotechnol. 2010 Feb 3;10:8.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.
Horseradishperoxidase conjugated Mouse anti Histidine tag antibody, clone AD1.1.10 (MCA1396P) used for the detection of HIS tagged protein constructs by western blotting.
Image caption:
Effect of host strain and induction conditions on periplasmic TCR expression.
(A) Representative western blots showing the expression profile of the TCR formats in either E. coli XL1-Blue (X) or RosettaBlue™ (R) where IPTG induction of protein expression either was omitted (-) or included (+). Normalized expression cultures were fractionated before the presence of TCR in the periplasmic fractions was analyzed by western blot detection using an anti-His-HRP antibody (n = 3). (B) The bacterial growth characteristics were monitored by measuring the OD600nm of E. coli XL1-blue (X) and RosettaBlue™ (R) bacterial cultures after overnight protein expression without (-) or with (+) IPTG induction at 30°C. n = 3 for all constructs except for controls (n = 4) and the dsTCR constructs (n = 1). Standard deviations are indicated where possible. (A, B) Cells expressing scFv anti-phOx were used as positive control and untransformed cells as negative control.
From: Gunnarsen KS, Høydahl LS, Neumann RS, Bjerregaard-Andersen K, Nilssen NR, Sollid LM, et al. (2018)
Soluble T-cell receptor design influences functional yield in an E. coli chaperone-assisted expression system.
PLoS ONE 13(4): e0195868.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.
Horseradishperoxidase conjugated Mouse anti Histidine tag antibody, clone AD1.1.10 (MCA1396P) used for the detection of HIS tagged protein constructs by western blotting.
Image caption:
Expression characteristics of the recombinant TCR formats.
Representative western blot showing the expression profile of the TCR formats. E. coli RosettaBlue™ expression cultures were normalized and fractionated into; M, medium; P, periplasmic; C, cytosolic fractions before analysis by western blots (n = 3–4). All samples were detected with anti-His-HRP antibody. Expression of scFv anti-phOx was included as positive control.
From: Gunnarsen KS, Høydahl LS, Neumann RS, Bjerregaard-Andersen K, Nilssen NR, Sollid LM, et al. (2018)
Soluble T-cell receptor design influences functional yield in an E. coli chaperone-assisted expression system.
PLoS ONE 13(4): e0195868.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.
Mouse anti Histidine Tag antibody, clone AD1.1.10 (MCA1396) used to detect LAG3 specific, HIS tagged antibody by western blotting.
Image caption:
Characterization of the anti-LAG3 scFvF7.
D.Binding analysis of scFvF7. 0.5 μg of both recombinant LAG3 and glucose oxidase (GO) proteins were loaded in each replicate well on a 12% SDS-PAGE under reducing condition and transferred to filter paper. Strips from the filter were then incubated with the indicted primary antibodies. Anti-6 his mAb was used as a positive control for LAG3 recombinant protein (which has a 6-histidines tag at its C terminal end). As additional negative controls, two strips were incubated respectively with the anti-Flag antibody and the HRP-conjugated anti-mouse antibody, in order to monitor eventual non-specific signals due to the secondary antibodies used to detect scFv antibodies. The ELISA shown below (performed with the same secondary Abs) is a check for the reactivity of the scFv-containing supernatants (scFvGO and scFvF7) used in the assay. Arrows indicate relevant signals. Molecular markers in kilodaltons (kDa) are reported on the right. A representative of three independent assays is shown.
From: Ascione A, Arenaccio C, Mallano A, Flego M, Gellini M, Andreotti M, Fenwick C, Pantaleo G, Vella S, Federico M.
Development of a novel human phage display-derived anti-LAG3 scFv antibody targeting CD8+ T lymphocyte exhaustion.
BMC Biotechnol. 2019 Oct 17;19(1):67.
doi: 10.1186/s12896-019-0559-x.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Histidine Tag antibody, clone AD1.1.10 (MCA1396) used to detect LAG3 specific, HIS tagged antibody by western blotting.
Image caption:
Construction and characterization of the divalent scFvF7-Fc Ab.
Anti 6 × histidine western blot analysis carried on supernatants of CHO cells transfected with either the scFvF7-Fc expressing vector (lane 1), or transfected with the empty vector (lane 2). Molecular weight markers (Mk) are reported on the left. The results are representative of four independent assays.
From: Ascione A, Arenaccio C, Mallano A, Flego M, Gellini M, Andreotti M, Fenwick C, Pantaleo G, Vella S, Federico M.
Development of a novel human phage display-derived anti-LAG3 scFv antibody targeting CD8+ T lymphocyte exhaustion.
BMC Biotechnol. 2019 Oct 17;19(1):67.
doi: 10.1186/s12896-019-0559-x.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Histidine tag antibody, clone AD1.1.10 (MCA1396) used to detect Histidine tagged spiHIFα by western blotting.
Image caption:
HEK293 cells were transfected with 0.1μg of spiHIFα or spiHIFβ plasmids. 24h post-transfection, cells were maintained in normoxic (21%) or hypoxic (<1%) conditions during 6h. Whole cell extracts migrated within a 7.5% SDS PAGE gel, were analyzed by immuno blot with different antibodies: anti-Histidine tag for SpiHIFα, anti-Myc for SpiHIFβ, and anti-actin as a loading control.
From: Zoccola D, Morain J, Pagès G, Caminiti-Segonds N, Giuliano S, Tambutté S, et al. (2017)
Structural and functional analysis of coral Hypoxia Inducible Factor.
PLoS ONE 12(11): e0186262.
doi: 10.1371/journal.pone.0186262.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Histidine tag antibody, clone AD1.1.10 (MCA1396) used to stain His tagged protein by westeern blotting
Image caption:
Immunoblot of purified protein targets of ubiquitylation by recombinant wild-type and Ser65Asp-ubiquitin in HEK293T cells. His FLAG-tagged UbWT and UbS65D mutant proteins were expressed in HEK293T cells. Following cell lysis in DLB buffer and sonication at 3 x 30 s on ice, cell fragments were pelleted at 13,000 g for 10 min. Proteins bound to UbWT and UbS65D were purified by IMAC, resolved by SDS-PAGE and immunoblotted against VU-1 ubiquitin (left panel) or anti-His-tag antibodies (right panel). Blots were stripped and reprobed with anti-β-actin antibodies (lower panels)
From: Dongdem JT, Dawson SP, Layfield R.
Modification of small ubiquitin-related modifier 2 (SUMO2) by phosphoubiquitin in HEK293T cells.
Proteomics. 2021 Aug;21(15):e2000234.
doi: 10.1002/pmic.202000234.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Histidine tag antibody, clone AD1.1.10 (MCA1396) used to identify alpaca derived nanobodies bearing a histidine tag by western blotting.
Image caption:
Western blot showing secretion and accumulation of Nbs conjugated to a pelB secretion signal by transformed S. glossinidius strains in spent media; Lane 1, MW marker, Lane 2, recSodalis::Nb_An46; Lane 3, recSodalis::Nb_88; Lane 4, recSodalis::Nb_19 and Lane 5, recSodalis::Nb_63.
Concentrated spent medium was tested using an anti-His antibody.
From: De Vooght L, De Ridder K, Hussain S, Stijlemans B, De Baetselier P, Caljon G, Van Den Abbeele J.
Targeting the tsetse-trypanosome interplay using genetically engineered Sodalis glossinidius. PLoS Pathog. 2022 Mar 10;18(3):e1010376.
doi: 10.1371/journal.ppat.1010376.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Filter by Application:
WB F ResetMouse anti Histidine Tag:DyLight®800
- Product Type
- Monoclonal Antibody
- Clone
- AD1.1.10
- Isotype
- IgG1
- Specificity
- Histidine Tag
Antibody Quality and Performance Guarantee
| Mouse anti Histidine tag antibody, clone AD1.1.10 recognizes proteins and peptides containing the motif H-H-H-H-H-H and is therefore of value in detecting proteins containing histidine tags. Clone AD1.1.10 has been used to detect and purify histidine-tagged proteins expressed in mammalian ( In Western blotting of bacterial extracts the antibody has been shown not to cross-react with any endogenous products, although some cross-reactivity may be seen with extracts of insect or mammalian cells. This antibody is routinely tested in Western blotting on histidine tagged recombinant proteins and reacts against all histidine-tagged proteins so far tested. |
- Target Species
- Synthetic Peptide
- Product Form
- Purified IgG conjugated to DyLight® 800 - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.09% sodium azide (NaN3)
- Immunogen
- PAX6 transcription factor linked to histidine tag.
- Approx. Protein Concentrations
- IgG concentration 1.0mg/ml
- Fusion Partners
- Spleen cells from immunized Balb/c mice were fused with cells of the mouse NS1 myeloma cell line.
- Max Ex/Em
-
Fluorophore Excitation Max (nm) Emission Max (nm) Dylight®800 777 794 - Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- Acknowledgements
- DyLight is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries. His-tag is a registered trademark of EMD Biosciences.
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. This product is photosensitive and should be protected from light.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. This product is photosensitive and should be protected from light.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| Western Blotting |  |
1/1000 | 1/5000 |
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
References for Histidine Tag antibody
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Ascione, A. et al. (2019) Development of a novel human phage display-derived anti-LAG3 scFv antibody targeting CD8+ T lymphocyte exhaustion.
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- RRID
- AB_11152596
MCA1396D800GA
153640
159645
165957
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