SKU PA1026-1
Size 100μg/vial
Reactivity Human, Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications IHC, ICC, WB

Overview

Product Name Anti-Connexin 43/GJA1 Antibody
SKU/Catalog Number PA1026-1
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.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for Gap junction alpha-1 protein(GJA1) detection. Tested with WB, IHC-P, IHC-F, ICC in Human;Mouse;Rat.
Cite This Product Anti-Connexin 43/GJA1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1026-1)
Host Rabbit
Immunogen A synthetic peptide corresponding to a sequence at the C-terminus of human Connexin 43(351-367aa HELQPLAIVDQRPSSRA), identical to the related rat and mouse sequences.
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Immunocytochemistry , 0.5-1μg/ml
Immunohistochemistry(Frozen Section), 0.5-1μg/ml
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml
Western blot, 0.1-0.5μg/ml

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), IHC(F) and ICC.

Images And Assay Conditions

/antibody/pa1026 1 1 IHC anti connexin 43 gja1 antibody.jpg

Figure 1. IHC analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026-1).
Connexin 43/GJA1 was detected in paraffin-embedded section of human lung cancer 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 (PA1026-1) 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.

/antibody/pa1026 1 2 IHC anti connexin 43 gja1 antibody.jpg

Figure 2. IHC analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026-1).
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 (PA1026-1) 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.

/antibody/pa1026 1 3 IHC anti connexin 43 gja1 antibody.jpg

Figure 3. IHC analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026-1).
Connexin 43/GJA1 was detected in frozen 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 (PA1026-1) 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.

/p/a/pa1026 1 4_1.jpg

Figure 4. Western blot analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026-1).
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 hear 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 # PA1026-1) 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.

/p/a/pa1026 1 5.jpg

Figure 5. IHC analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026-1).
Connexin 43/GJA1 was detected in paraffin-embedded section of human intestinal cancer 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 (PA1026-1) 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.

/p/a/pa1026 1 6.jpg

Figure 6. IHC analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026-1).
Connexin 43/GJA1 was detected in paraffin-embedded section of mouse 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 (PA1026-1) 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.

/p/a/pa1026 1 7.jpg

Figure 7. IHC analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026-1).
Connexin 43/GJA1 was detected in paraffin-embedded section of human mammary cancer 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 (PA1026-1) 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.

/p/a/pa1026 1 8.jpg

Figure 8. IHC analysis of Connexin 43/GJA1 using anti- Connexin 43/GJA1 antibody (PA1026-1).
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 (PA1026-1) 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.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P17302
Gene Name GJA1
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.

Ontology

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). .
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.

Order Product (PA1026-1)

Promotion:

Buy primary get secondary antibody for free.

$50 fee for conjugation. Antibody size is reduced to 50ug.

Option Price
30ug sample size $99
100ug $280
100ug+Free HRP Secondary BA1054 $280
100ug+Free Biotin Secondary BA1003 $280

USD $280

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Troubleshooting

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Publications

Effect of Novel Gasotransmitter hydrogen sulfide on renal fibrosis and connexins expression in diabetic rats
Argirein alleviates stress-induced and diabetic hypogonadism in rats via normalizing testis endothelin receptor A and connexin 43
miR-381suppresses C/EBP?-dependent Cx43 expression in breast cancer cells
Identification of miR-200a as a novel suppressor of connexin 43in breast cancer cells
The Compound Chinese Medicine ?Kang Fu Ling? Protects against High Power Microwave-Induced Myocardial Injury
CPU86017-RS attenuate hypoxia-induced testicular dysfunction in mice by normalizing androgen biosynthesis genes and pro-inflammatory cytokines
Engineered myocardial tissues constructed in vivo using cardiomyocyte-like cells derived from bone marrow mesenchymal stem cells in rats
The Compound Chinese Medicine ?Kang Fu Ling? Protects against High Power Microwave-Induced Myocardial Injury
Li H, Tang M, Liang H, Li Y, Wang J, Song Y, Zheng Y, Xi J, Zhang J, Hescheler J, Zhu M. Cell Physiol Biochem. 2013;32(1):53-63. Doi: 10.1159/000350124. Epub 2013 Jul 4. Coculture Of Embryonic Ventricular Myocytes And Mouse Embryonic Stem Cell Enh...
Zhang X, Gao Y, Dong J, Wang S, Yao B, Zhang J, Hu S, Xu X, Zuo H, Wang L, Zhou H, Zhao L, Peng R. Plos One. 2014 Jul 3;9(7):E101532. Doi: 10.1371/Journal.Pone.0101532. Ecollection 2014. The Compound Chinese Medicine "Kang Fu Ling" Protects Agains...
Peng Hj, Dai Dz, Ji H, Dai Y. Eur J Pharmacol. 2010 May 25;634(1-3):101-6. Doi: 10.1016/J.Ejphar.2010.02.001. Epub 2010 Feb 16. The Separate Roles Of Endothelin Receptors Participate In Remodeling Of Matrix Metalloproteinase And Connexin 43 Of Car...

Customer Q&As

Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
A: You can find the immunogen sequence under "Immunogen" and clonality in the product name.
Q: What is the expected band size? Why is it different than the observed band size?
A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:
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.,
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?
A: Some common names include but are not limited to connexin 43 antibody, connexin-43 antibody, connexin43 antibody, cx43 antibody, gja1 antibody