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, WB, IP, Flow Cyt
- Reacts with: Species independent
Overview
-
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, 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.
-
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.
-
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
-
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...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
JPAR-2 -
Isotype
IgG -
Research areas
Images
-
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.
-
Immunohistochemical analysis of agarose-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).
-
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.
-