Anti-AKAP12 Antibody Picoband™

Product Name

Anti-AKAP12 Antibody Picoband™

View all AKAP12 Antibodies

SKU/Catalog Number

A02303-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-AKAP12 Antibody Picoband™ catalog # A02303-3. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Mouse, Rat.

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-AKAP12 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02303-3)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived mouse AKAP12 recombinant protein (Position: E387-H1620).

*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

A02303-3 is reactive to AKAP12 in Mouse, Rat

Applications

A02303-3 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

300 kDa

Calculated molecular weight

191.482kDa

Background of AKAP12

A-kinase anchor protein 12, aka AKAP250, is an enzyme that in humans is encoded by the AKAP12 gene. The A-kinase anchor proteins (AKAPs) are a group of structurally diverse proteins, which have the common function of binding to the regulatory subunit of protein kinase A (PKA) and confining the holoenzyme to discrete locations within the cell. This gene encodes a member of the AKAP family. The encoded protein is expressed in endothelial cells, cultured fibroblasts, and osteosarcoma cells. It associates with protein kinase A and C and phosphatase, and serves as a scaffold protein in signal transduction. This protein and RII PKA colocalize at the cell periphery. This protein is a cell growth-related protein. Antibodies to this protein can be produced by patients with myasthenia gravis. Alternative splicing of this gene results in two transcript variants encoding different isoforms.

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.

View more details

Innovating Scientists Reward

If you are the first to review this product, or if you have results for a special sample, species or application this product is not validated in, share your results with us and receive product credits you can use towards any Boster products! Applicable to all scientists worldwide.

Submit A Review

Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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, 0.1-0.25μg/ml, Mouse
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse
Immunocytochemistry/Immunofluorescence, 5μg/ml, Mouse
Flow Cytometry, 1-3μg/1x10⁶ cells, Mouse, Rat
Direct ELISA, 0.1-0.5μg/ml, Mouse

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of AKAP12 using anti-AKAP12 antibody (A02303-3).
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: mouse brain tissue lysates,
Lane 2: mouse ovary 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-AKAP12 antigen affinity purified polyclonal antibody (Catalog # A02303-3) at 0.25 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 AKAP12 at approximately 300KD. The expected band size for AKAP12 is at 181KD.

Click image to see more details

Figure 2. IHC analysis of AKAP12 using anti-AKAP12 antibody (A02303-3).
AKAP12 was detected in paraffin-embedded section of mouse intestine 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-AKAP12 Antibody (A02303-3) 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. Flow Cytometry analysis of HEPA1-6 cells using anti-AKAP12 antibody (A02303-3).
Overlay histogram showing HEPA1-6 cells stained with A02303-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AKAP12 Antibody (A02303-3,1μg/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 (Red line) was also used as a control.

Click image to see more details

Figure 4. Flow Cytometry analysis of NRK cells using anti-AKAP12 antibody (A02303-3).
Overlay histogram showing NRK cells stained with A02303-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AKAP12 Antibody (A02303-3,1μg/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 (Red line) was also used as a control.

Click image to see more details

Figure 5. IF analysis of AKAP12 using anti-AKAP12 antibody (A02303-3).
AKAP12 was detected in immunocytochemical section of HEPA1-6 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-AKAP12 Antibody (A02303-3) 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.

Gene/Protein Information For AKAP12 (Source: Uniprot.org, NCBI)

Alternative Names

A kinase (PRKA) anchor protein (gravin) 12; A kinase (PRKA) anchor protein 12; AKAP 250; AKAP250250kDa; DKFZp686M0430; DKFZp686O0331; FLJ20945; FLJ97621; Gravin; kinase scaffold protein gravin; myasthenia gravis autoantigen gravin; Myasthenia gravis autoantigen AKAP12 AKAP250, SSeCKS A-kinase anchoring protein 12 A-kinase anchor protein 12|A kinase (PRKA) anchor protein 12|A-kinase anchor protein, 250kDa|AKAP 250|Src-Suppressed C Kinase Substrate|kinase scaffold protein gravin|myasthenia gravis autoantigen gravin

For more info on AKAP12, check out the AKAP12 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 AKAP12: 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].

Take me to the AKAP12 Infographic

Here are featured tools and databases that you might find useful.

• Boster's Pathways Library
• Protein Databases
• Bioscience Research Protocol Resources
• Data Processing & Analysis Software
• Photo Editing Software
• Scientific Literature Resources
• Research Paper Management Tools
• Molecular Biology Software
• Primer Design Tools
• Bioinformatics Tools
• Phylogenetic Tree Analysis