Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)

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® - BSA and Azide free
  • Suitable for: WB, IHC-P, IP, ELISA
  • Reacts with: Species independent

Overview

  • Product name

    Anti-6X His tag® antibody [3D5] - BSA and Azide free
    See all 6X His tag® primary antibodies

  • Description

    Mouse monoclonal [3D5] to 6X His tag® - BSA and Azide free

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, IHC-P, IP, ELISAmore details

  • Species reactivity

    Reacts with: Species independent

  • Immunogen

    Other Immunogen Type. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: His tagged 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®.

  • General notes

    ab255888 is the carrier-free version of ab206491. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab255888 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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
    For more information see here.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    pH: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    3D5

  • Isotype

    IgG2b

  • Light chain type

    kappa

  • Research areas

Images

  • Western blot - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)
    Western blot - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)
    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 seconds


    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252883).

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252883).

  • Western blot - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)
    Western blot - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)
    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 second


    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252883).

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)
    Immunoprecipitation - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252883).

  • ELISA - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)
    ELISA - Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252883).

  • Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)
    Anti-6X His tag® antibody [3D5] - BSA and Azide free (ab255888)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

© 2021 Astana Biomed Group LLP. All rights reserved.