Ultra-LEAF™ Purified anti-His Tag Antibody

Pricing & Availability
Clone
J095G46 (See other available formats)
Other Names
Hexa histidine-tag, 6xHis-tag, 6XHis, 6His
Isotype
Mouse IgG2a, κ
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Product Citations
publications
J095G46_Pure_His-tag_Antibody_1_110117
Total cell lysate (15 µg protein) from 293E (lane 1) and 293E transfected with His-tag fused protein (lane 2) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 1:1000 diluted (0.5 µg/mL), 1:2500 diluted (0.2 µg/mL) and 1:10000 diluted (0.05 µg/mL) purified anti-His tag antibody (clone J095G46). Proteins were visualized by chemiluminescence detection using 1:3000 diluted HRP Goat anti-mouse IgG Antibody (Cat. No. 405306, upper). 1:2000 dilution of Direct-Blot™ HRP anti-GAPDH antibody (clone FF26A/F9, Cat. No. 649203) was used as a loading control (lower). Lane M: MW ladder.
  • J095G46_Pure_His-tag_Antibody_1_110117
    Total cell lysate (15 µg protein) from 293E (lane 1) and 293E transfected with His-tag fused protein (lane 2) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 1:1000 diluted (0.5 µg/mL), 1:2500 diluted (0.2 µg/mL) and 1:10000 diluted (0.05 µg/mL) purified anti-His tag antibody (clone J095G46). Proteins were visualized by chemiluminescence detection using 1:3000 diluted HRP Goat anti-mouse IgG Antibody (Cat. No. 405306, upper). 1:2000 dilution of Direct-Blot™ HRP anti-GAPDH antibody (clone FF26A/F9, Cat. No. 649203) was used as a loading control (lower). Lane M: MW ladder.
  • J095G46_Pure_His-tag_Antibody_2_110117
    HeLa cells transiently transfected with His tag fused protein or non-transfected were fixed with 100% methanol for 10 minutes and blocked with 5% FBS for 60 minutes. Then the cells were stained overnight at 4 degrees with (A) 1: 500 diluted (1 µg/mL) mouse IgG2a isotype control (Cat. No. 401501) or, anti-His tag antibody at (B) 1: 500 (1 µg/mL), (C) 1: 1667 (0.3 µg/mL), (D) 1: 5000 (0.1 µg/mL) dilution, followed by 1:250 diluted (2 µg/mL) Alexa Fluor® 488 (green) conjugated goat anti-mouse IgG (Cat. No. 405319) antibody for one hour at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective.
  • J095G46_PURE_His-Tag_Antibody_FC_3_031418
    CL hTMEM8A transfected P3.U1 stained with purified anti-His Tag (clone J097D2) (filled histogram) or purified mouse IgG2a, κ isotype control (open histogram), followed by anti-mouse IgG PE.
Cat # Size Price Quantity Avail. Save
362615 100 µg £141
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362616 1 mg £403
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Description

This antibody was raised against a linear 8XHis tag. It reacts with His-tagged protein with C, N or internal 6XHis tags or longer. It can be used for western blotting to detect linear His repeats (6XHis or more). It can also be used for ICFC and ICC staining of His-tagged proteins.

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Human, Mouse, Rat, All species
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
KLH-conjugated His-tag peptide.
Formulation
0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative. Endotoxin level is <0.01 EU/µg of the protein (<0.001 ng/µg of the protein) as determined by the LAL test.
Preparation
The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration
1.0 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.
Application

WB - Quality tested
ICC, FC - Validated

Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 1:1000-1:10000 diluted (0.05 - 0.5 µg/mL). For immunocytochemistry, a concentration range of 1:500 - 1:5000 (0.1 - 1.0 µg/mL) is recommended. For flow cytometric staining, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This antibody was raised against a linear 8X His tag. It reacts with proteins displaying repeats of 6 Histidines (His6) or more in either internal or C and N terminal regions. It is suitable for ICFC.

Clone J095G46 has been validated for immunocytochemistry (ICC).

Antigen Details

Structure
6 repeat His amino acid
Function
Epitope tag
Biology Area
Cell Biology, Immunology
Antigen References
  1. Zhao C, et al. 2010. Anal Biochem. 399:237
  2. Salichs E, et al. 2009. PLoS Genet. 5:e1000397
  3. Terpe K. 2003. Appl. Microbiol. Biotechnol. 60:523
  4. Lindner, P, et al. 1997. Biotechniques. 22:140
  5. Burritt JB, et al. 1995. J. Biol. Chem. 270:16974.
  6. Taylor RM, et al. 2007. Methods Mol. Biol. 412:429.
Gene ID
NA
UniProt
View information about His Tag on UniProt.org

Related FAQs

Does BioLegend test each LEAF™ antibody by functional assay?

No, BioLegend does not test LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.

Do you guarantee that your antibodies are totally pathogen free?

BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.

Does BioLegend test each LEAF™ antibody for potential pathogens?

No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.

Have you tested this LEAF™/Ultra-LEAF™ antibody for in vivo or in vitro applications?

We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.

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For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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