Recombinant Anti-TIM44 antibody [EPR16821-26] (ab201453)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16821-26] to TIM44
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP
- Reacts with: Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-TIM44 antibody [EPR16821-26]
See all TIM44 primary antibodies -
Description
Rabbit monoclonal [EPR16821-26] to TIM44 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IPmore details -
Species reactivity
Reacts with: Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: K562, Daudi, HepG2, PC-12 and C6 cell lysates; Human fetal liver lysate. ICC/IF: K562 and HepG2 cells. IP: K562 whole cell lysate.
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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
EPR16821-26 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab201453 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 |
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Flow Cyt (Intra) |
1/500.
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WB | (1) |
1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 51 kDa).
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ICC/IF |
1/500.
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IP |
1/120.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 51 kDa). |
ICC/IF
1/500. |
IP
1/120. |
Target
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Relevance
TIM44 is the essential component of the PAM complex, a complex required for the translocation of transit peptide-containing proteins from the inner membrane into the mitochondrial matrix in an ATP-dependent manner. It recruits mitochondrial HSP70 to drive protein translocation into the matrix using ATP as an energy source. -
Cellular localization
Mitochondrion inner membrane. -
Database links
- Entrez Gene: 10469 Human
- Entrez Gene: 29635 Rat
- Omim: 605058 Human
- SwissProt: O43615 Human
- SwissProt: O35094 Rat
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Alternative names
- DKFZp686H05241 antibody
- MIMT44 antibody
- Mitochondrial inner membrane translocase antibody
see all
Images
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Intracellular Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) labelling TIM44 with purified ab201453 at 1/500 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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All lanes : Anti-TIM44 antibody [EPR16821-26] (ab201453) at 1/2000 dilution
Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma) cell lysate
Lane 3 : Daudi (Human Burkitt's lymphoma cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 51 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-TIM44 antibody [EPR16821-26] (ab201453) at 1/10000 dilution + Human fetal liver lysate at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 51 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-TIM44 antibody [EPR16821-26] (ab201453) at 1/2000 dilution
Lane 1 : C6 (Rat glial tumor cells) cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 51 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling TIM44 with ab201453 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasm staining on HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab201453 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling TIM44 with ab201453 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasm staining on K562 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab201453 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
TIM44 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab201453 at 1/120 dilution. Western blot was performed from the immunoprecipitate using ab201453 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.
Lane 1: K562 whole cell lysate 10 µg (Input). Lane 2: ab201453 IP in K562 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab201453 in K562 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 1 second.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab201453 has not yet been referenced specifically in any publications.