Recombinant
RabMAb

Recombinant Anti-Optineurin antibody [EPR20654] (ab213556)

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Overview

  • Product name

    Anti-Optineurin antibody [EPR20654]
    See all Optineurin primary antibodies

  • Description

    Rabbit monoclonal [EPR20654] to Optineurin

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, IHC-P, WB, ICC/IF, IHC-Frmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human Optineurin aa 50-350. The exact sequence is proprietary.
    Database link: Q96CV9

  • 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; 293T and NIH/3T3 whole cell lysates. IHC-P: Human retina and cerebrum tissues; Mouse retina and cerebrum tissues; Rat cerebellum tissue. IP: 293T whole cell lysate.

  • 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
    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

Our Abpromise guarantee covers the use of ab213556 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
IP 1/40.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 66 kDa).
ICC/IF 1/50.
IHC-Fr 1/100.

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] (ab213556)
    Immunohistochemistry (Frozen sections) - Anti-Optineurin antibody [EPR20654] (ab213556)

    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).

  • Western blot - Anti-Optineurin antibody [EPR20654] (ab213556)
    Western blot - Anti-Optineurin antibody [EPR20654] (ab213556)
    All lanes : Anti-Optineurin antibody [EPR20654] (ab213556) at 1/1000 dilution

    Lane 1 : Human skeletal muscle lysate at 20 µg
    Lane 2 : 293T (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
    Lane 3 : Rat brain lysate at 20 µg
    Lane 4 : Mouse retina lysate at 20 µg
    Lane 5 : Rat retina lysate at 20 µg
    Lane 6 : Rat placenta lysate at 20 µg
    Lane 7 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 66 kDa
    Observed band size: 68 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-3/7: 3 minutes; Lane 4-6: 10 seconds.

    The MW observed is consistent with the literature (PMID 20174559, PMID 24983867, PMID 11834836).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] (ab213556)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] (ab213556)

    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.

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

  • Immunocytochemistry/ Immunofluorescence - Anti-Optineurin antibody [EPR20654] (ab213556)
    Immunocytochemistry/ Immunofluorescence - Anti-Optineurin antibody [EPR20654] (ab213556)

    ab213556 staining Optineurin in the NIH/3T3 (mouse embryonic fibroblast) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde, permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/50). ab150077 Alexa Fluor® 488 Goat anti-Rabbit secondary (1/1000) was used as the secondary antibody. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (1/200) was used as a counterstain. 

  • Immunohistochemistry (Frozen sections) - Anti-Optineurin antibody [EPR20654] (ab213556)
    Immunohistochemistry (Frozen sections) - Anti-Optineurin antibody [EPR20654] (ab213556)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-Optineurin antibody [EPR20654] (ab213556)
    Immunocytochemistry/ Immunofluorescence - Anti-Optineurin antibody [EPR20654] (ab213556)

    ab213556 staining Optineurin in the HeLa (human cervix adenocarcinoma epithelial cell) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde, permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/50). ab150077 Alexa Fluor® 488 Goat anti-Rabbit secondary (1/1000) was used as the secondary antibody. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (1/200) was used as a counterstain. 

  • Western blot - Anti-Optineurin antibody [EPR20654] (ab213556)
    Western blot - Anti-Optineurin antibody [EPR20654] (ab213556)
    All lanes : Anti-Optineurin antibody [EPR20654] (ab213556) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal kidney lysate
    Lane 3 : Human placenta lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Predicted band size: 66 kDa
    Observed band size: 68 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 seconds; Lane 2/3: 30 seconds.

    The MW observed is consistent with the literature (PMID 20174559, PMID 24983867, PMID 11834836).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] (ab213556)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] (ab213556)

    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.

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

  • Western blot - Anti-Optineurin antibody [EPR20654] (ab213556)
    Western blot - Anti-Optineurin antibody [EPR20654] (ab213556)
    All lanes : Anti-Optineurin antibody [EPR20654] (ab213556) at 1/1000 dilution

    Lane 1 : Rat heart lysate
    Lane 2 : Mouse placenta lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 66 kDa
    Observed band size: 68 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 minutes; Lane 2: 10 seconds.

    The MW observed is consistent with the literature (PMID 20174559, PMID 24983867, PMID 11834836).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] (ab213556)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] (ab213556)

    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.

    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] (ab213556)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] (ab213556)

    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.

    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] (ab213556)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Optineurin antibody [EPR20654] (ab213556)

    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.

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

  • Immunoprecipitation - Anti-Optineurin antibody [EPR20654] (ab213556)
    Immunoprecipitation - Anti-Optineurin antibody [EPR20654] (ab213556)

    Optineurin was immunoprecipitated from 0.35 mg of 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 1/10,000 dilution

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

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

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

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

    Exposure time: 3 minutes.

References

Publishing research using ab213556? Please let us know so that we can cite the reference in this datasheet.

ab213556 has been referenced in 1 publication.

  • Hu S  et al. Structure of Myosin VI/Tom1 complex reveals a cargo recognition mode of Myosin VI for tethering. Nat Commun 10:3459 (2019). PubMed: 31371777

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