Immunofluorescence analysis of CONNEXIN 31 was performed using 90% confluent log phase SH-SY5Y 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 Connexin 31 Rabbit Polyclonal Antibody (Product # 36-5100) at 2µg/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 conjµgate (Product # 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 (Product # 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 cell junction localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the C-terminal region of mouse connexin 31 protein|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (PFA fixed) (IHC (PFA))||1-3 µg/mL|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1-3 µg/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.
This antibody reacts with the ~32 kDa mouse Connexin 31 on Western blots; the band observed at ~62 kDa is probably due to dimerization of Connexin 31. Several higher bands of unknown origin are observed in western blots of mouse skin homogenates.
Gap junctions are conduits that allow the direct cell-to-cell passage of small cytoplasmic molecules, including ions, metabolic intermediates, and second messengers, and thereby mediate intercellular metabolic and electrical communication. Gap junction channels consist of connexin protein subunits, which are encoded by a multigene family. GJBs (gap-junction proteins or connexins) play crucial functional roles associated with these channels. Defects in GJB3 have been linked to erythrokeratodermia variabilis (EKV) is an autosomal dominant genodermatosis characterized by transient figurate red patches or hyperkeratosis. Mutations in GJB2 have also been associated with genetically derived hearing impairments, including autosomal recessive nonsyndromic deafness.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Death of Neurons following Injury Requires Conductive Neuronal Gap Junction Channels but Not a Specific Connexin.
36-5100 was used in western blot to determine if Cx36 gap junctions contribute to neuronal death via channel-dependent or channel-independent mechanisms.
|Fontes JD,Ramsey J,Polk JM,Koop A,Denisova JV,Belousov AB||PloS one (10:null)||2015|
|Human||1:200||Connexin expression and gap junctional coupling in human cumulus cells: contribution to embryo quality.||Wang HX,Tong D,El-Gehani F,Tekpetey FR,Kidder GM||Journal of cellular and molecular medicine (13:972)||2009|
||Connexin expression and gap junctional coupling in human cumulus cells: contribution to embryo quality.||Wang HX,Tong D,El-Gehani F,Tekpetey FR,Kidder GM||Journal of cellular and molecular medicine (13:972)||2009|