Anti-6X His tag® antibody [3D5] (ab252883)
![Anti-6X His tag® antibody [3D5] (ab252883)](https://www.abcam.com/ps/products/252/ab252883/Images/ab252883-395277-anti-6x-his-tag-antibody-3d5-western-blot-histagged-mouse-cathepsin-d-recombinant-protein.jpg "Anti-6X His tag® antibody [3D5] (ab252883)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [3D5] to 6X His tag®
- Suitable for: ELISA, WB, IHC-P, IP
- Reacts with: Species independent
Overview
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Product name
Anti-6X His tag® antibody [3D5]
See all 6X His tag® primary antibodies -
Description
Mouse monoclonal [3D5] to 6X His tag® -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species 
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Immunogen
Other Immunogen Type. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: His tagged recombinant proteins. IHC-P: HEK-293T transfected with a 6X His tag® construct. IP: HEK-293T transfected with cas9 (S. Aureas) expression vector containing a myc-His-tag®.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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
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, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
3D5 -
Isotype
IgG2b -
Light chain type
kappa -
Research areas
Images
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Western blot - Anti-6X His tag® antibody [3D5] (ab252883)All lanes : Anti-6X His tag® antibody [3D5] (ab252883) at 0.995 µg/ml
Lane 1 : His-tagged mouse Cathepsin D recombinant protein (aa 65-310; 27kDa)
Lane 2 : His-tagged mouse CDKN2A/p16INK4a recombinant protein (aa 1-168;18kDa)
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [3D5] (ab252883)](https://www.abcam.com/ps/products/252/ab252883/Images/ab252883-395275-ab252883395275anti6xhistagantibody3d5immunohistochemistryahek293ttransfectedwithaa6histagconstructbhek293ttransfectedwithemptyplasmid.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [3D5] (ab252883)Immunohistochemical analysis of paraffin-embedded (Panel A) HEK-293T transfected with a 6X His tag® construct and (Panel B) HEK-293T transfected with empty plasmid, labeling 6X His tag® with ab252883 at 1/2000 dilution (0.498µg/ml) followed by ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879). Positive staining on (Panel A) HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a 6X His tag® construct, no staining on (Panel B) HEK-293T transfected with empty plasmid. The section was incubated with ab252883 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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![Western blot - Anti-6X His tag® antibody [3D5] (ab252883)](https://www.abcam.com/ps/products/252/ab252883/Images/ab252883-395276-anti-6x-his-tag-antibody-3d5-western-blot-hek293t.jpg)
Western blot - Anti-6X His tag® antibody [3D5] (ab252883)All lanes : Anti-6X His tag® antibody [3D5] (ab252883) at 0.995 µg/ml
Lane 1 : HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293T transfected with human histone H3.1 expression vector containing a myc-His-tag®, whole cell lysate
Lane 3 : HEK-293T transfected with human GAPDH expression vector containing a myc-His-tag®, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
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![Immunoprecipitation - Anti-6X His tag® antibody [3D5] (ab252883)](https://www.abcam.com/ps/products/252/ab252883/Images/ab252883-395274-anti-6x-his-tag-antibody-3d5-immunoprecipitation-hek293t-transfected-with-cas9.jpg)
Immunoprecipitation - Anti-6X His tag® antibody [3D5] (ab252883)6X His tag® was immunoprecipitated from HEK-293T transfected with cas9 (S. Aureas) expression vector containing a myc-His-tag®, whole cell lysate with ab252883 at 1/30 dilution (33.167µg/ml) (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252883 at 1/1000 dilution (0.995µg/ml). VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HEK-293T transfected with cas9 (S. Aureas) expression vector containing a myc-His-tag®, whole cell lysate 10µg.
Lane 2: ab252883 IP in HEK-293T transfected with cas9 (S. Aureas) expression vector containing a myc-His-tag®, whole cell lysate.
Lane 3: Mouse monoclonal IgG2b instead of ab252883 in HEK-293T transfected with cas9 (S. Aureas) expression vector containing a myc-His-tag®, whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 mins.
This blot was developed using a higher sensitivity ECL substrate.
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![ELISA - Anti-6X His tag® antibody [3D5] (ab252883)](https://www.abcam.com/ps/products/252/ab252883/Images/ab252883-395278-anti-6x-his-tag-antibody-3d5-elisa.jpg)
ELISA - Anti-6X His tag® antibody [3D5] (ab252883)ELISA - Anti-6X His tag® antibody [3D5] (ab252883).
ab252883 used at 0-5000 ng/ml.
Antigen (6X His tag®) concentration 1000 ng/ml.
Secondary is Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Mouse IgG (H+L) used at a 1/1000 dilution.
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![Anti-6X His tag® antibody [3D5] (ab252883)](https://www.abcam.com/ps/products/252/ab252883/Images/ab252883-6-benefits-of-recombinant-antibodies.png)
Anti-6X His tag® antibody [3D5] (ab252883)