Recombinant Anti-6X His tag® antibody [JPAR-2] (ab245114)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [JPAR-2] to 6X His tag®
- Suitable for: IHC-P, WB, IP, Flow Cyt
- Reacts with: Species independent
Related conjugates and formulations
Overview
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Product name
Anti-6X His tag® antibody [JPAR-2]
See all 6X His tag® primary antibodies -
Description
Rabbit monoclonal [JPAR-2] to 6X His tag® -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, IP, Flow Cytmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Species independent -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa 1-1053; 125kDa) construct, whole cell lysate. IHC-P: HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5) construct. Flow cyt: HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5) construct.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
JPAR-2 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab245114 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000.
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IP |
1/30.
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Flow Cyt |
1/500.
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Notes |
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IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. |
IP
1/30. |
Flow Cyt
1/500. |
Target
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Relevance
The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. His-tags can be fused to the amino- or carboxy- termini of proteins in transfected or transformed cells. -
Cellular localization
Depends upon the localization of the parent protein tagged with hexahistidine. -
Alternative names
- 6 His epitope tag antibody
- Hexa His tag antibody
- HHHHHH epitope tag antibody
see all
Images
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His-tagged Staphylococcus aureus cas9 (J7RUA5; aa 1-1053; 125kDa) was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) (transfected with Staphylococcus aureus cas9 (J7RUA5; aa 1-1053; 125kDa)) whole cell lysate with ab245114 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245114 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 whole cell lysate 10 μg (Input).
Lane 2: ab245114 IP in HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245114 in HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds. -
Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5) construct labeling 6X His tag® with ab245114 at 1/500 (Right) compared with a Rabbit monoclonal IgG isotype control (ab172730) (Left). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079), at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemical analysis of paraffin-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) labeling 6X His tag® with ab245114 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on (A) HEK-293T transfected with a 6× His tag construct is observed. No staining on (B) HEK-293T transfected with an empty plasmid is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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All lanes : Anti-6X His tag® antibody [JPAR-2] (ab245114) at 1/1000 dilution
Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty plasmid, whole cell lysate
Lane 2 : HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa 1-1053; 125kDa) construct, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 125 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking and dilution buffer: 5% NFDM/TBST.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab245114 has been referenced in 1 publication.
- Vadla GP et al. ABT1 modifies SMARD1 pathology via interactions with IGHMBP2 and stimulation of ATPase and helicase activity. JCI Insight 8:N/A (2023). PubMed: 36480289