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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

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Immunohistochemistry (Frozen sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Western blot - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Immunoprecipitation - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Immunohistochemistry (Frozen sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Immunoprecipitation - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
  • Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20654] to Optineurin - BSA and Azide free
  • Suitable for: IHC-Fr, IP, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-Optineurin antibody [EPR20654] - BSA and Azide free
    See all Optineurin primary antibodies

  • Description

    Rabbit monoclonal [EPR20654] to Optineurin - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-Fr, IP, WB, IHC-Pmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal brain, fetal kidney, placenta and skeletal muscle lysates; Rat brain, retina, placenta and heart lysates; Mouse retina and placenta lysates; U-2 OS, 293T and NIH/3T3 whole cell lysates. IHC-P: Human retina and cerebrum tissues; Mouse retina and cerebrum tissues; Rat cerebellum tissue. IP: 293T and U-2 OS whole cell lysate.

  • General notes

    ab240979 is the carrier-free version of ab213556.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab240979 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
IHC-Fr
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 68 kDa (predicted molecular weight: 66 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
IHC-Fr
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 68 kDa (predicted molecular weight: 66 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for ICC/IF.

Target

  • Function

    Plays an important role in the maintenance of the Golgi complex, in membrane trafficking, in exocytosis, through its interaction with myosin VI and Rab8. Links myosin VI to the Golgi complex and plays an important role in Golgi ribbon formation. Negatively regulates the induction of IFNB in response to RNA virus infection. Plays a neuroprotective role in the eye and optic nerve. Probably part of the TNF-alpha signaling pathway that can shift the equilibrium toward induction of cell death. May act by regulating membrane trafficking and cellular morphogenesis via a complex that contains Rab8 and hungtingtin (HD). May constitute a cellular target for adenovirus E3 14.7, an inhibitor of TNF-alpha functions, thereby affecting cell death.

  • Tissue specificity

    Present in acqueous humor of the eye (at protein level). Highly expressed in trabecular meshwork. Expressed nonpigmented ciliary epithelium, retina, brain, adrenal cortex, fetus, lymphocyte, fibroblast, skeletal muscle, heart, liver, brain and placenta.

  • Involvement in disease

    Defects in OPTN are the cause of primary open angle glaucoma type 1E (GLC1E) [MIM:137760]. Primary open angle glaucoma (POAG) is characterized by a specific pattern of optic nerve and visual field defects. The angle of the anterior chamber of the eye is open, and usually the intraocular pressure is increased. The disease is asymptomatic until the late stages, by which time significant and irreversible optic nerve damage has already taken place.
    Defects in OPTN are a cause of susceptibility to normal pressure glaucoma (NPG) [MIM:606657].
    Defects in OPTN are the cause of amyotrophic lateral sclerosis type 12 (ALS12) [MIM:613435]. It is a neurodegenerative disorder affecting upper motor neurons in the brain and lower motor neurons in the brain stem and spinal cord, resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology of amyotrophic lateral sclerosis is likely to be multifactorial, involving both genetic and environmental factors. The disease is inherited in 5-10% of the cases.

  • Domain

    Ubiquitin-binding motif (UBAN) is essential for its inhibitory function, subcellular localization and interaction with TBK1.

  • Post-translational
    modifications

    Phosphorylated. Phosphorylation is induced by phorbol esters and decreases its half-time.

  • Cellular localization

    Cytoplasm > perinuclear region. Golgi apparatus. Golgi apparatus > trans-Golgi network. Found in the perinuclear region and associates with the Golgi apparatus. Colocalizes with MYO6 and RAB8 at the Golgi complex and in vesicular structures close to the plasma membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 14.7K interacting protein antibody
    • Ag9 C5 antibody
    • ALS12 antibody
    • E3 14.7K interacting protein antibody
    • E3-14.7K-interacting protein antibody
    • FIP 2 antibody
    • FIP-2 antibody
    • FIP2 antibody
    • Glaucoma 1 open angle E (adult onset) antibody
    • Glaucoma 1 open angle E antibody
    • GLC1E antibody
    • HIP 7 antibody
    • HIP-7 antibody
    • HIP7 antibody
    • Huntingtin interacting protein 7 antibody
    • Huntingtin interacting protein HYPL antibody
    • Huntingtin interacting protein L antibody
    • Huntingtin yeast partner L antibody
    • Huntingtin-interacting protein 7 antibody
    • Huntingtin-interacting protein L antibody
    • HYPL antibody
    • Injury inducible protein I 55 antibody
    • NEMO related protein antibody
    • NEMO-related protein antibody
    • NRP antibody
    • Optic neuropathy inducing protein antibody
    • Optic neuropathy-inducing protein antibody
    • Optineurin antibody
    • OPTN antibody
    • OPTN_HUMAN antibody
    • TFIIIA IntP antibody
    • TFIIIA-IntP antibody
    • Transcription factor IIIA interacting protein antibody
    • Transcription factor IIIA-interacting protein antibody
    • Tumor necrosis factor alpha inducible cellular protein containing leucine zipper domains antibody
    see all

Images

  • Immunohistochemistry (Frozen sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Immunohistochemistry (Frozen sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

    ab213556 staining Optineurin in Mouse retina tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with 4% paraformaldehyde, permeabilized with 0.2% Triton. Samples were incubated with primary antibody (1/100). An Alexa Fluor® 488 Goat anti-Rabbit secondary (1/1000) was used as the secondary antibody. Counter stained with DAPI.

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

    Positive staining on mouse retina (PMID: 15607428).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213556).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

    Immunohistochemical analysis of paraffin-embedded human retina tissue labeling Optineurin with ab213556 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human retina (PMID: 15607428).

    Counter stained 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213556).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Western blot - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    All lanes : Anti-Optineurin antibody [EPR20654] (ab213556) at 1/5000 dilution

    Lane 1 : Wild-type U-2 OS cell lysate
    Lane 2 : OPTN knockout U-2 OS cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 66 kDa



    This data was developed using the same antibody in a different buffer formulation (ab213556). 

    ab213556 was shown to react with OPTN in wild-type U-2 OS cells in Western blot with loss of signal observed in a OPTN knockout cell line. Wild-type U-2 OS and OPTN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab213556 overnight at 4 °C at a 1/5000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 1/5000 before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

  • Immunoprecipitation - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Immunoprecipitation - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    This data was developed using the same antibody clone in a different buffer formulation (ab240979). Immunoprecipitation of OPTN in U-2 OS cells. Lysates were prepared and immunoprecipitation was performed using 1.0 μg of ab213556 pre-coupled to prot.A-Sepharose beads. Samples were washed and processed for western blot with ab213556 at 1/5000. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitate. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
  • Immunohistochemistry (Frozen sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Immunohistochemistry (Frozen sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

    ab213556 staining Optineurin in Rat retina tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with 4% paraformaldehyde, permeabilized with 0.2% Triton. Samples were incubated with primary antibody (1/100). An Alexa Fluor® 488 Goat anti-Rabbit secondary (1/1000) was used as the secondary antibody. Counter stained with DAPI.

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

    Positive staining on rat retina (PMID: 15607428).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213556).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

    Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Optineurin with ab213556 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human cerebrum (PMID: 26303227).

    Counter stained 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213556)

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

    Immunohistochemical analysis of paraffin-embedded mouse retina tissue labeling Optineurin with ab213556 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on mouse retina (PMID: 15607428).

    Counter stained 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213556)

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

    Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Optineurin with ab213556 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on mouse cerebrum (PMID: 26303227).

    Counter stained 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213556)

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

    Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling Optineurin with ab213556 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on rat cerebellum (PMID: 26303227).

    Counter stained 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213556)

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Immunoprecipitation - Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

    Optineurin was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab213556 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab213556 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293T whole cell lysate 10 μg (Input).

    Lane 2: ab213556 IP in HEK-293T whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213556 in HEK-293T whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213556)

  • Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)
    Anti-Optineurin antibody [EPR20654] - BSA and Azide free (ab240979)

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.

Click here to view the general protocols

References (0)

Publishing research using ab240979? Please let us know so that we can cite the reference in this datasheet.

ab240979 has not yet been referenced specifically in any publications.

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