Synthetic peptide conjugated to KLH derived from within residues 1650 to the C-terminus of Human Clathrin heavy chain.
(Peptide available as ab23440.)
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.
应用 | Ab评论 | 说明 |
---|---|---|
WB | Use a concentration of 1 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 180 kDa). | |
ICC/IF | Use a concentration of 1 µg/ml. | |
IHC-P | Use at an assay dependent concentration. | |
Flow Cyt | Use at an assay dependent concentration. ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
|
IHC-FoFr | Use at an assay dependent concentration. PubMed: 18852301 | |
IP | Use at an assay dependent concentration. |
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).
Immunocytochemical analysis of Mouse bEnd.3 labeling Clathrin heavy chain with ab21679 at 1/300 dilution using secondary anti-rabbit-alexa fluor 488 at 1/500 dilution
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.
ab21679 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.
Further results for this antibody can be found at www.proteinatlas.org
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%)
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%)
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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"