Anti-6X His tag® antibody [HIS.H8] (ab18184)
Overview
- Product nameAnti-6X His tag® antibody [HIS.H8]See all 6X His tag® primary antibodies ...
- DescriptionMouse monoclonal [HIS.H8] to 6X His tag®
- SpecificityRecognizes His-tagged recombinant proteins or His-tagged proteins overexpressed in cells.
- Tested applicationsICC/IF, ELISA, Dot Blot, WB, IP, EMSA more details
- Immunogen
6x His synthetic peptide.
- General notesHIS-TAG® is a trademark of EMD Biosciences, Inc.
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.1% Sodium Azide
Constituents: PBS, pH 7.2 -
Concentration information loading... - PurityIgG fraction
- Clonality Monoclonal
- Clone numberHIS.H8
- IsotypeIgG2b
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab18184 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| ICC/IF | ICC/IF: 1/200. |
| ELISA | ELISA: Use at an assay dependent dilution. |
| Dot Blot | Dot: Use at an assay dependent dilution. |
| WB | WB: 1/1000. |
| IP | IP: 1/200. |
| EMSA | EMSA: Use at an assay dependent dilution. PubMed: 20606007 |
Target
- RelevanceThe H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. The antibody recognizes the His-tag fused to the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.
- Cellular localizationDepends upon the localization of the parent protein tagged with hexahistidine.
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Alternative names
- 6 His epitope tag antibodyHexa His tag antibodyHHHHHH epitope tag antibody
- HHHHHH tag antibodyHis tag antibody
see all
Anti-6X His tag® antibody [HIS.H8] images
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Immunofluorescence (red) with ab18184 on His-tag fusion protein in 293 cells.
Left hand panel: untransfected control
Right hand panel: transfected.
Counterstained with DAPI (blue). -
All lanes : Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/1000 dilution
Lane 1 : 293 cells, untransfected control
Lane 2 : 293 cells transfected with His-tagged vector -
Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/5000 dilution + Conditioned medium from transfected HEK293 cells which produce recombinant secretory protein with a 6XHis-Tag at 5 µg
Secondary
An HRP-conjugated Horse anti mouse polyclonal at 1/5000 dilutionThis image is courtesy of an anonymous Abreview
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Immunocytochemistry/ Immunofluorescence - 6X His tag® antibody [HIS.H8] (ab18184)This image is courtesy of an anonymous abreview.ab18184 at a 1/1000 dilution staining 6X His tag in transfected human HEK293 cells by Immunocytochemistry/ Immunofluorescence incubated for 12 hours at 22°C. PFA fixed. Permeabilized using 0.1% Triton X-100 in PBS. Blocked using 3% BSA for 1 hour at 22°C. Secondary used at a 1/1000 dilution polyclonal Goat anti-mouse IgG (H+L) conjugated to Alexa Fluor 488 (green). DAPI staining (blue). -
Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184)This image is courtesy of an Abreview submitted by Mr Chris TracyImmunoprecipitation of His/FLAG tagged PDCD5 using ab18184 at 1/100 dilution.
Lane 1 : MW Ladder (from top 120, 85, 50, 34, 25, 19 kDa)
Lanes 2-5 : Immunoprecipitation of His/FLAG tagged PDCD5 using ab18184 from mixture of ~50 ng purified His/FLAG tagged PDCD5 and 15 µL RRL in 300 µL buffer.
Lanes 6-7 : Purified PDCD5 from E. coli with ~60-70 ng protein.
The lowest band in each lane is ~ 20 kDa and is PDCD5. The upper bands in lanes 2-5 are the heavy chain (~50 kDa, runs as doublet) and light chain (~25 or 26 kDa).
Western Blot was performed with a mouse anti-FLAG primary antibody at 1/2000 dilution. An IRDye®800CW-conjugated goat anti-mouse IgG was used as a secondary antibody.
References for Anti-6X His tag® antibody [HIS.H8] (ab18184)
This product has been referenced in:
- Panter MS et al. Dynamics of Major Histocompatibility Complex Class I Association with the Human Peptide-loading Complex. J Biol Chem 287:31172-84 (2012). WB . Read more (PubMed: 22829594) »
- Cojocaru M et al. Transcription Factor IIS Cooperates with the E3 Ligase UBR5 to Ubiquitinate the CDK9 Subunit of the Positive Transcription Elongation Factor B. J Biol Chem 286:5012-22 (2011). Read more (PubMed: 21127351) »
![Immunocytochemistry/ Immunofluorescence - 6X His tag®, antibody [HIS.H8] (ab18184) Immunocytochemistry/ Immunofluorescence - 6X His tag®, antibody [HIS.H8] (ab18184)](http://a.abcam.com/ps/datasheet/Images/18/ab18184/ab18184_2.jpg)
![Western blot - 6X His tag antibody [HIS.H8] (ab18184) Western blot - 6X His tag antibody [HIS.H8] (ab18184)](http://a.abcam.com/ps/datasheet/Images/18/ab18184/ab18184_3.jpg)
![Western blot - 6X His tag® antibody [HIS.H8] (ab18184) Western blot - 6X His tag® antibody [HIS.H8] (ab18184)](http://a.abcam.com/ps/datasheet/images/18/ab18184/6X-His-tag-Primary-antibodies-ab18184-2.jpg)