Immunofluorescence analysis of KCNQ3 was performed using 70% confluent log phase RSC-96 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with KCNQ3 Rabbit Polyclonal Antibody (PA1-930) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||GST fusion protein encoding the first 71 amino acids of rat KCNQ3.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.02% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Frozen) (IHC (F))||0.5 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Miscellaneous PubMed (MISC)||See 1 publications below|
PA1-930 detects KCNQ3 from mouse, human and rat samples. This antibody is specific for KCNQ3 and does not detect KCNQ1, KCNQ2, KCNQ4 or KCNQ5.
PA1-930 has been successfully used in immunofluorescence, immunohistochemistry and immunocytochemistry procedures. Immunohistochemical staining using PA1-930 yielded a strong signal mainly in interneurons and astrocytes in the dentate region of rat hippocampal samples.
PA1-930 immunogen is a GST fusion protein encoding the first 71 amino acids of rat KCNQ3.
KCNQs are members of the voltage-dependent non-inactivating potassium channel family. Currently there are five known KCNQs (KCNQ1-5) found in the central nervous system and KCNQ2 and 3 have demonstrated their importance in M-current activation.
Studies have shown that KCNQ2 and KCNQ3 form heteromultimers that, when formed, substantially increase the M-current. Inhibition of M-current controls neuron excitability throughout the nervous system as well as the responsiveness to synaptic inputs. Genetic mutations in these proteins have been linked to disorders such as benign familial neonatal convulsions (BFNC), deafness, neuropathic pain and epilepsy.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Antibodies and a cysteine-modifying reagent show correspondence of M current in neurons to KCNQ2 and KCNQ3 K+ channels.
PA1-930 was used in immunocytochemistry and immunohistochemistry to investigate the influence of KCNQ2 and KCNQ3 potassium channels on the M current of neurons
|Roche JP,Westenbroek R,Sorom AJ,Hille B,Mackie K,Shapiro MS||British journal of pharmacology (137:1173)||2002|