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.
Key Product Details
Species Reactivity
Validated:
Multi-Species
Cited:
Human, Mouse, Bovine, Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque), Yeast
Applications
Validated:
Western Blot
Cited:
Immunohistochemistry-Frozen, Binding Assay, Bioassay, ELISA Capture, ELISA Detection, ELISA Development, Epitope Mapping
Label
Biotin
Antibody Source
Monoclonal Mouse IgG1 Clone # AD1.1.10
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Product Specifications
Immunogen
His-tagged peptide
Specificity
Detects proteins containing accessible consecutive histidine regions. This antibody detects His tags localized at the amino- or carboxyl-terminus.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1
Scientific Data Images for His Tag Biotinylated Antibody
Detection of His Tag by Western Blot.
Western blot shows lysates of HEK293T 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 BAI-1. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-His Tag Biotinylated Monoclonal Antibody (Catalog # BAM050) followed by Streptavidin-HRP (Catalog # DY998) in 3% BSA. 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.
Applications for His Tag Biotinylated Antibody
Application
Recommended Usage
Western Blot
0.1 µg/mL
Sample: HEK293T human embryonic kidney cell line and CHO Chinese hamster ovary cell line transfected with His-tagged proteins
Sample: HEK293T human embryonic kidney cell line and CHO Chinese hamster ovary cell line transfected with His-tagged proteins
Reviewed Applications
Read 6 reviews rated 4.2 using BAM050 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 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.
Calculators
Background: His Tag
Long Name
Histidine Tag
Alternate Names
polyHistidine
Additional His Tag Products
Product Documents for His Tag Biotinylated Antibody
Product Specific Notices for His Tag Biotinylated Antibody
For research use only
Citations for His Tag Biotinylated Antibody
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Customer Reviews for His Tag Biotinylated Antibody (6)
4.2 out of 5
6 Customer Ratings
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Application: Western BlotSample Tested: Cell LysatesSpecies: MouseVerified Customer | Posted 11/07/2021
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Application: Simple WesternSample Tested: Purified proteinSpecies: HumanVerified Customer | Posted 06/20/2018
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Application: Affinity PurificationSample Tested: antigen specific B cellsSpecies: RabbitVerified Customer | Posted 12/01/2017
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Application: Western BlotSample Tested: SerumSpecies: MouseVerified Customer | Posted 05/16/2016
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Application: Western BlotSample Tested: Cell LysatesSpecies: E. coliVerified Customer | Posted 02/23/2016Antibody was used to probe cell lysates from E. coli to identify protein of interest which carries a C-terminal his-tag. The yield of our protein is very low in the soluble fraction, so we used this mAb to compare the soluble fraction to the inclusion body prep to see if there was a signal in the inclusion body prep indicating that our protein may not be soluble.
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Application: biosensor capture assay.Sample Tested: Monoclonal Mouse IgG1 Clone # AD1.1.10Species: MouseVerified Customer | Posted 02/22/2016This antibody was used to capture his tagged proteins in various biosensor assays. This antibody works fine for general purposes. In some platforms (i.e., Octet), it is not as robust as a capture reagent, but it is still acceptable.
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for His Tag Biotinylated Antibody
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A: 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.