Product Info Summary
SKU: | A01708-3 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-cGKI/PRKG1 Antibody Picoband®
View all cGK1/PRKG1 Antibodies
SKU/Catalog Number
A01708-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-cGKI/PRKG1 Antibody Picoband® catalog # A01708-3. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-cGKI/PRKG1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01708-3)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human cGKI/PRKG1 recombinant protein (Position: S2-Q44).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A01708-3 is reactive to PRKG1 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
78 kDa
Calculated molecular weight
76.364kDa
Background of cGK1/PRKG1
cGMP-dependent protein kinase 1, alpha isozyme is an enzyme that in humans is encoded by the PRKG1 gene. Mammals have three different isoforms of cyclic GMP-dependent protein kinase (Ialpha, Ibeta, and II). These PRKG isoforms act as key mediators of the nitric oxide/cGMP signaling pathway and are important components of many signal transduction processes in diverse cell types. This PRKG1 gene on human chromosome 10 encodes the soluble Ialpha and Ibeta isoforms of PRKG by alternative transcript splicing. A separate gene on human chromosome 4, PRKG2, encodes the membrane-bound PRKG isoform II. The PRKG1 proteins play a central role in regulating cardiovascular and neuronal functions in addition to relaxing smooth muscle tone, preventing platelet aggregation, and modulating cell growth. This gene is most strongly expressed in all types of smooth muscle, platelets, cerebellar Purkinje cells, hippocampal neurons, and the lateral amygdala. Isoforms Ialpha and Ibeta have identical cGMP-binding and catalytic domains but differ in their leucine/isoleucine zipper and autoinhibitory sequences and therefore differ in their dimerization substrates and kinase enzyme activity.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A01708-3 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human, Rat
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: rat lung tissue, rat heart tissue, mouse lung tissue, human HEK293 whole cell
IHC: mouse lung tissue, rat lung tissue, hman breast cancer tissue
ICC/IF: U20S cell
FCM: RH35 cell, U251 cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of CGKI/PRKG1 using anti-CGKI/PRKG1 antibody (A01708-3).
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: rat lung tissue lysates,
Lane 2: rat heart tissue lysates,
Lane 3: mouse lung tissue lysates,
Lane 4: human HEK293 whole cell 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-CGKI/PRKG1 antigen affinity purified polyclonal antibody (Catalog # A01708-3) 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 1:5000 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 CGKI/PRKG1 at approximately 78KD. The expected band size for CGKI/PRKG1 is at 78KD.
Click image to see more details
Figure 2. IHC analysis of CGKI/PRKG1 using anti-CGKI/PRKG1 antibody (A01708-3).
CGKI/PRKG1 was detected in paraffin-embedded section of mouse lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-CGKI/PRKG1 Antibody (A01708-3) 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.
Click image to see more details
Figure 3. IHC analysis of CGKI/PRKG1 using anti-CGKI/PRKG1 antibody (A01708-3).
CGKI/PRKG1 was detected in paraffin-embedded section of rat lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-CGKI/PRKG1 Antibody (A01708-3) 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.
Click image to see more details
Figure 4. IHC analysis of CGKI/PRKG1 using anti-CGKI/PRKG1 antibody (A01708-3).
CGKI/PRKG1 was detected in paraffin-embedded section of hman breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-CGKI/PRKG1 Antibody (A01708-3) 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.
Click image to see more details
Figure 5. IF analysis of CGKI/PRKG1 using anti-CGKI/PRKG1 antibody (A01708-3).
CGKI/PRKG1 was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-CGKI/PRKG1 Antibody (A01708-3) 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. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 6. Flow Cytometry analysis of RH35 cells using anti-CGKI/PRKG1 antibody (A01708-3).
Overlay histogram showing RH35 cells stained with A01708-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CGKI/PRKG1 Antibody (A01708-3, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Click image to see more details
Figure 7. Flow Cytometry analysis of U251 cells using anti-CGKI/PRKG1 antibody (A01708-3).
Overlay histogram showing U251 cells stained with A01708-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CGKI/PRKG1 Antibody (A01708-3, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For PRKG1 (Source: Uniprot.org, NCBI)
Gene Name
PRKG1
Full Name
cGMP-dependent protein kinase 1
Weight
76.364kDa
Superfamily
protein kinase superfamily
Alternative Names
E3 SUMO-protein ligase PIAS1; DEAD/H box-binding protein 1; E3 SUMO-protein transferase PIAS1; Gu-binding protein; GBP; Protein inhibitor of activated STAT protein 1; RNA helicase II-binding protein; PIAS1; DDXBP1 PRKG1 AAT8, PKG, PKG1B, PRKGR1B, cGK, cGK 1, cGK1, cGKI, cGKI-BETA, cGKI-alpha, PRKG1 protein kinase cGMP-dependent 1 cGMP-dependent protein kinase 1|protein kinase, cGMP-dependent, regulatory, type I, beta|protein kinase, cGMP-dependent, type I
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on PRKG1, check out the PRKG1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for PRKG1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-cGKI/PRKG1 Antibody Picoband® (A01708-3)
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