Recombinant

Recombinant Anti-iNOS antibody [EPR16635-ChiM-IgG2b] (ab210823)

Overview

  • Product name

    Anti-iNOS antibody [EPR16635-ChiM-IgG2b]
    See all iNOS primary antibodies
  • Description

    Mouse monoclonal [EPR16635-ChiM-IgG2b] to iNOS
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Human iNOS aa 1-200. The exact sequence is proprietary.
    Database link: P35228

  • Positive control

    • WB: RAW 264.7 whole cell lysate treated with 0.1 µg/mL LPS for 6 hours. ICC-IF: Raw264.7 cells untreated and LPS treated.
  • General notes

    This mouse antibody has been engineered from a RabMAb parent antibody (ab178945). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.

Properties

Applications

Our Abpromise guarantee covers the use of ab210823 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
WB Use a concentration of 5 µg/ml. Detects a band of approximately 131 kDa (predicted molecular weight: 131 kDa).
ICC/IF Use a concentration of 1 - 5 µg/ml.

Target

  • Function

    Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In macrophages, NO mediates tumoricidal and bactericidal actions. Also has nitrosylase activity and mediates cysteine S-nitrosylation of cytoplasmic target proteins such COX2.
  • Tissue specificity

    Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.
  • Sequence similarities

    Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
  • Information by UniProt
  • Database links

  • Alternative names

    • HEP-NOS antibody
    • Hepatocyte NOS antibody
    • HEPNOS antibody
    • inducible antibody
    • Inducible nitric oxide synthase antibody
    • Inducible NO synthase antibody
    • Inducible NOS antibody
    • iNOS antibody
    • MAC NOS antibody
    • Macrophage NOS antibody
    • Nitric oxide synthase 2 inducible antibody
    • Nitric oxide synthase 2 inducible macrophage antibody
    • nitric oxide synthase 2A (inducible, hepatocytes) antibody
    • Nitric oxide synthase antibody
    • Nitric oxide synthase inducible antibody
    • nitric oxide synthase, macrophage antibody
    • NOS 2 antibody
    • NOS antibody
    • Nos II antibody
    • NOS type II antibody
    • nos2 antibody
    • NOS2_HUMAN antibody
    • NOS2A antibody
    • NOS2A, Inducible, Hepatocyte antibody
    • Peptidyl-cysteine S-nitrosylase NOS2 antibody
    see all

Images

  • ab210823 stained in Raw264.7 cells. Untreated and LPS treated (1ug/ml, 24 hours) cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab210823 at 5µg/ml overnight at +4°C then detected with an Alexa Fluor® 488 goat anti-mouse secondary antibody (ab150117) at a 1/1000 dilution (shown in green). ab206369 (Rabbit monoclonal [EPR16774] to beta Tubulin Alexa Fluor® 594) was used as a counterstaining at a 1/200 dilution for 1hour at room temperature (pseudo-colored red). DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • Lane 1 : Anti-iNOS antibody [EPR16635-ChiM-IgG2b] (ab210823) at 5 µg
    Lane 2 : Anti-iNOS antibody [EPR16635] (ab178945) at 5 µg

    All lanes : RAW 264.7 whole cell lysate treated with 0.1 µg/mL LPS for 6 hours

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
    Lane 2 : Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 131 kDa
    Observed band size: 131 kDa


    Exposure time: 8 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab210823 (lane 1) and ab178945 (lane 2) overnight at 4°C. Antibody binding was detected using an anti-mouse (lane 1) and anti-rabbit (lane 2) antibody conjugated to HRP, and visualised using ECL development solution ab133406.

References

ab210823 has not yet been referenced specifically in any publications.

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