Recombinant Anti-Clathrin heavy chain antibody [EPR24236-222] (ab305259)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24236-222] to Clathrin heavy chain
- Suitable for: WB, ICC/IF, Flow Cyt (Intra), IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Clathrin heavy chain antibody [EPR24236-222]
See all Clathrin heavy chain primary antibodies -
Description
Rabbit monoclonal [EPR24236-222] to Clathrin heavy chain -
Host species
Rabbit -
Specificity
The antibody detects both heavy chain 1 and 2.
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Tested applications
Suitable for: WB, ICC/IF, Flow Cyt (Intra), IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa and HEK-293T whole cell lysate. ICC/IF: HeLa cells. Flow Cyt (Intra): HeLa cells. IP: HeLa 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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24236-222 -
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 ab305259 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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WB |
1/1000. Detects a band of approximately 180 kDa.
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ICC/IF |
1/50.
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Flow Cyt (Intra) |
1/500.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 180 kDa. |
ICC/IF
1/50. |
Flow Cyt (Intra)
1/500. |
IP
1/30. |
Target
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Function
Clathrin is the major protein of the polyhedral coat of coated pits and vesicles. Two different adapter protein complexes link the clathrin lattice either to the plasma membrane or to the trans-Golgi network. -
Sequence similarities
Belongs to the clathrin heavy chain family. -
Cellular localization
Cytoplasmic vesicle membrane. Membrane > coated pit. Melanosome. Cytoplasmic face of coated pits and vesicles. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Database links
- Entrez Gene: 1213 Human
- Omim: 118955 Human
- SwissProt: P53675 Human
- SwissProt: Q00610 Human
- Unigene: 491351 Human
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Alternative names
- CHC17 antibody
- Clathrin heavy chain antibody
- Clathrin heavy chain 1 antibody
see all
Images
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All lanes : Anti-Clathrin heavy chain antibody [EPR24236-222] (ab305259) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : 293T (human embryonic kidney epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/2000 dilution
Observed band size: 180 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 37 seconds
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Clathrin heavy chain was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg with ab305259 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305259 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2: ab305259 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab305259 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 50 seconds
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Flow cytometric (Intracellular) analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling Clathrin heavy chain with ab305259 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Clathrin heavy chain with ab305259 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing punctuated cytoplasmic staining in HeLa cell line. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab305259 has not yet been referenced specifically in any publications.