|Product Name||Anti-Connexin 43/GJA1 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Gap junction alpha-1 protein( GJA1) detection. Tested with WB, IHC-P, IF in Human;Mouse;Rat.|
|Cite This Product||Anti-Connexin 43/GJA1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1026)|
|Contents/Buffer||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Immunogen||A synthetic peptide corresponding to a sequence at the C-terminus of human Connexin 43(366-382aa RASSRASSRPRPDDLEI), identical to the related rat and mouse sequences.|
|Reactivity||Human, Mouse, Rat|
Assay Dilutions Overview
Immunofluorescence, 2μg/ml, Rat
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
Figure 1. Western blot analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat heart tissue lysates,
Lane 4: mouse brain tissue lysates,
Lane 5: mouse heart tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- Connexin 43/GJA1 antigen affinity purified polyclonal antibody (Catalog # PA1026) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Connexin 43/GJA1 at approximately 43KD. The expected band size for Connexin 43/GJA1 is at 43KD.
Figure 2. IHC analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026).
Connexin 43/GJA1 was detected in paraffin-embedded section of rat cardiac muscle tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- Connexin 43/GJA1 Antibody (PA1026) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 3. IF analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026).
Connexin 43/GJA1 was detected in paraffin-embedded section of rat heart tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- Connexin 43/GJA1 Antibody (PA1026) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C.. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Gap junction alpha-1 protein|
|Tissue Specificity||Expressed in the heart and fetal cochlea. .|
|Alternative Names||Gap junction alpha-1 protein;Connexin-43;Cx43;Gap junction 43 kDa heart protein;GJA1;GJAL;|
|Subcellular Localization||Cell membrane ; Multi-pass membrane protein . Cell junction, gap junction . Endoplasmic reticulum .|
|Molecular Weight||43008 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Gap junction protein that acts as a regulator of bladder capacity. A gap junction consists of a cluster of closely packed pairs of transmembrane channels, the connexons, through which materials of low MW diffuse from one cell to a neighboring cell. May play a critical role in the physiology of hearing by participating in the recycling of potassium to the cochlear endolymph. Negative regulator of bladder functional capacity: acts by enhancing intercellular electrical and chemical transmission, thus sensitizing bladder muscles to cholinergic neural stimuli and causing them to contract (By similarity). .|
|Research Areas||Atherosclerosis, Cardiovascular, Cell Adhesion, Cytoskeleton / Ecm, Signal Transduction, Vascular Inflammation
*You can search these to find other products in these research areas.
|Background||Connexins 43(Cx43), also called GAP Junction Protein, alpha-1(GJA1). Connexin 43 is a member of the connexin gene family which abundantly expressed in the heart and liver and was mapped to 6q21-q23.2. Connexin43, the major protein of gap junctions in the heart, is targeted by several protein kinases that regulate myocardial cell-cell coupling. Mutations in the connexin43 gap-junction gene, which lead to abnormally regulated cell-cell communication, are associated with visceroatrial heterotaxia. Cx43 must also play a critical role in the physiology of hearing, presumably by participating in the recycling of potassium to the cochlear endolymph.|
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1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,