In human HEK293 cells ab21089 recognised a band of ~58 kDa, the predicted MW of Kv1.6 (data not shown). This band was not blocked by the immunising peptide. Additional bands of 100 kDa, 25 kDa and 20 kDa were also recognised.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
1/40. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
Mediates the voltage-dependent potassium ion permeability of excitable membranes. Assuming opened or closed conformations in response to the voltage difference across the membrane, the protein forms a potassium-selective channel through which potassium ions may pass in accordance with their electrochemical gradient.
Belongs to the potassium channel family. A (Shaker) (TC 1.A.1.2) subfamily. Kv1.6/KCNA6 sub-subfamily.
The N-terminus may be important in determining the rate of inactivation of the channel while the tail may play a role in modulation of channel activity and/or targeting of the channel to specific subcellular compartments. The segment S4 is probably the voltage-sensor and is characterized by a series of positively charged amino acids at every third position.
All lanes : Anti-Kv1.6 antibody (ab21089) at 1 µg/ml
Lane 1 : Mouse Brain Lysate Lane 2 : Mouse Brain Lysate with Human Kv1.6 peptide (ab23431) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary Lane 1 : Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution Lane 2 : Alexa Fluor Goat polyclonal to Rabbit IgG
Predicted band size: 58 kDa
ab21089 recognises a band corresponding to the predicted MW of Kv1.6 in mouse brain lysate (lane 1). Antibody binding to this band may be blocked using the immunising peptide, ab23431 (lane 2). The antibody also recognises additional proteins. These may be degradation/cleavage products of Kv1.6, or they may represent non-specific binding by the antibody.
ICC/IF image of ab21089 stained human HEK 293 cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab21089, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
ab21089 staining Kv6.1 in human pancreas tissue. Paraffin embedded human pancreas tissue was incubated with ab21089 (1/40 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab21089 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
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Kv1.6 antibody (ab21089)This image is courtesy of an abreview submitted by Sophie Pezet, ESPCI, France
The IHC-FoFr image stainig Kv1.6(ab21089) on Rat DRG sections. The sections used came from animals perfused fixed with Paraformaldehyde 4%, in phosphate buffer 0.2M. Following postfixation in the same fixative overnight, the tissues were cryoprotected in sucrose 30% overnight. Tissues were then cut using a cryostat.