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
Overview
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Product name
Anti-TMC1 antibody [EPR19527]
See all TMC1 primary antibodies -
Description
Rabbit monoclonal [EPR19527] to TMC1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-Fr MouseIP HumanWB Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19527 -
Isotype
IgG -
Research areas
Images
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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.
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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.
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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 secondBlocking/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.
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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.
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