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    tmc1-antibody-epr19527-ab199949.pdf

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RecombinantRabMAb

Recombinant Anti-TMC1 antibody [EPR19527] (ab199949)

  • Datasheet
  • Certificate of Compliance
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Immunocytochemistry/ Immunofluorescence - Anti-TMC1 antibody [EPR19527] (ab199949)
  • Immunohistochemistry (Frozen sections) - Anti-TMC1 antibody [EPR19527] (ab199949)
  • Western blot - Anti-TMC1 antibody [EPR19527] (ab199949)
  • Immunoprecipitation - Anti-TMC1 antibody [EPR19527] (ab199949)
  • Anti-TMC1 antibody [EPR19527] (ab199949)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19527] to TMC1
  • Suitable for: WB, IHC-Fr, ICC/IF, IP
  • Reacts with: Mouse, Human

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Overview

  • Product name

    Anti-TMC1 antibody [EPR19527]
    See all TMC1 primary antibodies
  • Description

    Rabbit monoclonal [EPR19527] to TMC1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Fr, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK-293T whole cell lysate transfected with DDDDK tagged mouse TMC1 . IHC-Fr: P20 mouse cochlea tissue. ICC/IF: Mouse TMC1-GFP transfected HEK-293T cells. IP: HEK-293T whole cell lysate transfected with pcDNA3.1-TMC1.
  • 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

  • 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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19527
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Sensory System
    • Auditory system

Associated products

  • Alternative Versions

    • Anti-TMC1 antibody [EPR19527] - Low endotoxin, Azide free (ab223541)
    • Anti-TMC1 antibody [EPR19527] - BSA and Azide free (ab271966)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab199949 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
1/2000. Detects a band of approximately 87 kDa (predicted molecular weight: 88 kDa).
IHC-Fr
1/50.
ICC/IF
1/500.
IP
1/30.
Notes
WB
1/2000. Detects a band of approximately 87 kDa (predicted molecular weight: 88 kDa).
IHC-Fr
1/50.
ICC/IF
1/500.
IP
1/30.

Target

  • Function

    Probable ion channel required for the normal function of cochlear hair cells.
  • Tissue specificity

    Detected in fetal cochlea, and at low levels in placenta and testis.
  • Involvement in disease

    Deafness, autosomal dominant, 36
    Deafness, autosomal recessive, 7
  • Sequence similarities

    Belongs to the TMC family.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession Q8TDI8 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 117531 Human
    • Entrez Gene: 13409 Mouse
    • Omim: 606706 Human
    • SwissProt: Q8TDI8 Human
    • SwissProt: Q8R4P5 Mouse
    • Unigene: 670211 Human
    • Unigene: 219585 Mouse
    • Alternative names

      • 4933416G09Rik antibody
      • Beethoven protein antibody
      • Bth antibody
      • CWEA1 antibody
      • Deafness autosomal dominant 36 antibody
      • Deafness autosomal recessive 11 antibody
      • Deafness autosomal recessive 7 antibody
      • Deafness protein antibody
      • DFNA36 antibody
      • DFNB11 antibody
      • DFNB7 antibody
      • dn antibody
      • Tmc1 antibody
      • TMC1_HUMAN antibody
      • Transmembrane channel like protein 1 antibody
      • Transmembrane channel-like protein 1 antibody
      • Transmembrane cochlear expressed protein 1 antibody
      • Transmembrane cochlear-expressed protein 1 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-TMC1 antibody [EPR19527] (ab199949)
      Immunocytochemistry/ Immunofluorescence - Anti-TMC1 antibody [EPR19527] (ab199949)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells, transfected with mouse TMC1-GFP or GFP only, labeling TMC1 with ab199949 at 1/500 dilution, followed by Alexa Fluor® 594 Goat anti-Rabbit (ab150080) secondary antibody at 1/1000 dilution (red).

      The results show cytoplasmic staining on mouse TMC1-GFP transfected HEK-293T cells.

      The nuclear counter stain is DAPI (blue).

      Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150115 (Alexa Fluor® 647 Goat anti-Mouse secondary (white)) at 1/1000 dilution.


      The negative controls are as follows:
      -ve control 1: ab199949 at 1/500 dilution followed by ab150115 (Alexa Fluor® 647 Goat anti-Mouse secondary (white)) at 1/1000 dilution.
      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150080 (Alexa Fluor® 594 Goat anti-Rabbit secondary) at 1/1000 dilution.

      Mouse TMC1 plasmid was kindly provided by our collaborator Dr. Yun Shi, Nanjing University MARC.

    • Immunohistochemistry (Frozen sections) - Anti-TMC1 antibody [EPR19527] (ab199949)
      Immunohistochemistry (Frozen sections) - Anti-TMC1 antibody [EPR19527] (ab199949)

      Immunohistochemical analysis of 4% paraformaldehyde-fixed frozen P20 mouse cochlea tissue labeling TMC1 with ab199949 at 1/50 dilution, followed by Goat Anti-Rabbit IgG (H+L) Dylight 488 secondary antibody at 1/500 dilution (green).

      The results show specific staining on cochlear hair cells.

      This data was kindly provided by our collaborator Dr. Yun Shi Nanjing University MARC.

      This is wholemount IF with no permeabilisation or antigen retrieval.

       

    • Western blot - Anti-TMC1 antibody [EPR19527] (ab199949)
      Western blot - Anti-TMC1 antibody [EPR19527] (ab199949)
      All lanes : Anti-TMC1 antibody [EPR19527] (ab199949) at 1/2000 dilution

      Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with empty DDDDK tag expression vector
      Lane 2 : HEK-293T whole cell lysate transfected with DDDDK tagged mouse TMC1

      Lysates/proteins at 2 µg per lane.

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

      Predicted band size: 88 kDa
      Observed band size: 87 kDa why is the actual band size different from the predicted?


      Exposure time: 1 second


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Anti-DDDDK tag antibody used is ab1162.

      Mouse TMC1 plasmid was kindly provided by our collaborator Dr. Yun Shi, Nanjing University MARC.

      WB detection of endogenous TMC1 is not shown due to unavailability of appropriate testing material. 

    • Immunoprecipitation - Anti-TMC1 antibody [EPR19527] (ab199949)
      Immunoprecipitation - Anti-TMC1 antibody [EPR19527] (ab199949)

      TMC1 was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate, transfected with pcDNA3.1-TMC1, with ab199949 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab199949 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 transfected with pcDNA3.1-TMC1, 10 μg (Input).

      Lane 2: ab199949 IP in HEK-293T whole cell lysate transfected with pcDNA3.1-TMC1.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199949 in HEK-293T whole cell lysate transfected with pcDNA3.1-TMC1.

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

      Exposure time: 1 second.

      Mouse TMC1 plasmid was kindly provided by our collaborator Dr. Yun Shi, Nanjing University MARC.

    • Anti-TMC1 antibody [EPR19527] (ab199949)
      Anti-TMC1 antibody [EPR19527] (ab199949)

    Protocols

    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (1)

    Publishing research using ab199949? Please let us know so that we can cite the reference in this datasheet.

    ab199949 has been referenced in 1 publication.

    • Yamaguchi S  et al. A Mechanosensitive Channel, Mouse Transmembrane Channel-Like Protein 1 (Mtmc1) Is Translated from a Splice Variant mTmc1ex1 but Not from the Other Variant mTmc1ex2. Int J Mol Sci 21:N/A (2020). PubMed: 32899784

    Customer reviews and Q&As

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