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Anti-Clathrin heavy chain 抗體 (ab21679)產(chǎn)品信息：
- Product name:Anti-Clathrin heavy chain antibody
- Description
Rabbit polyclonal to Clathrin heavy chain
- Tested applicationsWB, ICC/IF, IHC-P, Flow Cyt, IHC-FoFr, IP
- Species reactivityReacts with: Mouse, Rat, Hamster, Human
Predicted to work with: ChickenDoes not react with
African Green Monkey - Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1650 to the C-terminus of Human Clathrin heavy chain.
(Peptide available as ab23440.)
- Positive control
- This antibody gave a positive signal in HeLa, Jurkat, A431 & NIH3T3 cells, as well as human kidney tissues sections.
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 - Concentration Batch dependent within range: 100 µg at 0.3 - 1 mg/ml
- PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
- Signal Transduction
- Protein Trafficking
- Vesicle Transport
- Coat Proteins
- Tags & Cell Markers
- Subcellular Markers
- Organelles
- Caveolae and Clathrin
Applications
Our Abpromise guarantee covers the use of ab21679 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximay 180 kDa (predicted molecular weight: 180 kDa).Can be blocked with Human Clathrin heavy chain peptide (ab23440). |
| ICC/IF | ICC/IF: Use a concentration of 1 µg/ml. |
| IHC-P | IHC-P: Use at an assay dependent concentration. |
| Flow Cyt | Flow Cyt: Use at an assay dependent concentration. |
| IHC-FoFr | IHC-FoFr: Use at an assay dependent dilution. PubMed: 18852301 |
| IP | IP: Use at an assay dependent concentration. |
Target
- FunctionClathrin 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 similaritiesBelongs to the clathrin heavy chain family.
- Cellular localizationCytoplasmic 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
Database links
- Entrez Gene: 1213 Human
- Entrez Gene: 67300 Mouse
- Entrez Gene: 54241 Rat
- Omim: 118955 Human
- SwissProt: P53675 Human
- SwissProt: Q00610 Human
- SwissProt: Q68FD5 Mouse
- SwissProt: P11442 Rat
Alternative names
- CHC17 antibody
- Clathrin heavy chain 1 antibody
- Clathrin heavy chain 2 antibody
Anti-Clathrin heavy chain antibody images
Immunocytochemistry/ Immunofluorescence - Clathrin heavy chain antibody (ab21679)
ICC/IF image of ab21679 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21679, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Western blot - Clathrin heavy chain antibody (ab21679)
All lanes : Anti-Clathrin heavy chain antibody (ab21679) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Cytoplasmic lysate (ab14873)
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680 at 1/15000 dilution
Predicted band size : 180 kDa
Observed band size : 180 kDa
Immunoprecipitation - Clathrin heavy chain antibody (ab21679)This image is courtesy of an anonymous Abreview
ab21679 immunoprecipitating clathrin heavy chain from human HeLa S3 whole cell lysate (20ug). A protein A matrix was used with the antibody at a concentration of 4ug/mg of lysate. ab21679 was further used in the western blot stage at a dilution of 1/250. The image shows:
Lane 1: Clathrin IP (ab21679)
Lane 2: Control IP
Lane 3: Input (10%)
Immunoprecipitation - Clathrin heavy chain antibody (ab21679)This image is courtesy of an anonymous Abreview
ab21679 immunoprecipitating clathrin heavy chain from rat PC12 whole cell lysate (20ug). A protein A matrix was used with the antibody at a concentration of 4ug/mg of lysate. ab21679 was further used in the western blot stage at a dilution of 1/250. The image shows:
Lane 1: Clathrin IP (ab21679)
Lane 2: Control IP
Lane 3: Input (10%)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Clathrin heavy chain antibody (ab21679)
Image courtesy of Human Protein Atlasab21679 staining Clathrin heavy chain in human kidney. Paraffin embedded human (male) kidney tissue was incubated with ab21679 (1/7000 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab21679 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.
Flow Cytometry - Clathrin heavy chain antibody (ab21679)This image is a courtesy of Anonymous Abreview
ab21679 staining Clathrin heavy chain in mouse embryonic fibroblast cells by Flow Cytometery. Samples were prepared by homogenization and centrifugation in trypsin and they were washed in PBS. Cells were fixed in 70% 40C ethanol and cell population was gated on Non-junk by FSC/SSC. The primary antibody was diluted 1/2000 in 1% BSA/PBS and incubated with sample for 1 hour at 22°C. An Alexa Fuor 488 conjugated goat polyclonal to rabbit IgG (H&L) was used as secondary antibody at dilution at 1/500.
Immunocytochemistry/ Immunofluorescence - Clathrin heavy chain antibody (ab21679)This image is a courtesy of Anonymous Abreview
ab21679 staining Clathrin heavy chain in mouse embryonic fibroblasts by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde and permeabilized with -200C ethanol. Samples were incubated with primary antibody at 1/500 dilution (in 0.1%Saponin/1% BSA/PBS) for 1 hour. An Alexa Fluor®546-conjugated goat polyclonal to rabbit IgG (H+L) was used as secondary antibody at 1/500 dilution. Green color in image show positive staining with Alexa Fluor®546 conjugated secondary.
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