This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
PRKACB / cAMP-dependent protein kinase catalytic subunit beta · Western blot design guide
Design a Western Blot for PRKACB
Real validated PRKACB Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PRKACB WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
PRKACB Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~40.6 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 10 isoform(s) |
Section 1
Real Curated PRKACB Western Blot Protocols
The A05366-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A05366-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A05366-1) |
| Primary antibody | A05366-1 · 1:500 (catalog A05366-1) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A05366-1) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A05366-1) |
Section 2
What Is the Expected PRKACB Western Blot Band Size?
PRKACB is predicted at 40.6 kDa; splice isoforms and phosphorylation could affect bands, but their migration effects are not demonstrated.
| Band near 40.6 kDa | Consistent with the predicted PRKACB mass; confirm identity with antibody controls |
| Additional bands at different positions | Could reflect PRKACB splice isoforms, whose migration has not been established |
| Close doublet | Could reflect the listed phosphorylation states; a visible shift is not established |
| Lower band | Could represent a splice isoform or a fragment; identity needs confirmation |
💡Expected PRKACB appearancePRKACB has a predicted mass of 40.6 kDa; no empirical band size or resolvable isoform pattern is supplied, so confirm any candidate band with ordinary identity controls.
| UniProt predicted mass | Places the reference protein near 40.6 kDa before any migration effects |
| Splice isoforms 1, 2 and 3 | May differ in apparent size; individual masses and migration are unknown |
| Splice isoforms 4, 5 and 6 | May differ in apparent size; individual masses and migration are unknown |
| Splice isoforms 7, 8, 9 and 10 | May differ in apparent size; individual masses and migration are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | An isoform or phosphorylation-dependent mobility change is possible, but neither is established | Compare with an independent PRKACB antibody and a knockdown control |
| Band lower than expected | An isoform or proteolytic fragment is possible | Use a knockdown control and prepare fresh lysate with protease inhibitors |
| Multiple bands | Ten named splice isoforms and multiple phosphorylation sites are annotated | Check band identity by knockdown and compare phosphorylation-sensitive bands after phosphatase treatment |
| Weak or no signal | PRKACB detection may depend on sample amount or antibody performance | Check loading and antibody performance with a positive control lysate |
| Fragments below expected size | Proteolysis during sample preparation is possible | Prepare fresh chilled lysate with protease inhibitors and confirm PRKACB identity |
Experiment evidence
Western blot images showing PRKACB band
A05366-1 · PRKACB
A05366-1 · PRKACB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PRKACB Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PRKACB in Western blot, you can use appendix tissue, which HPA rates as highly positive.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA lists no undetected tissue, so use siRNA knockdown or a KO line for a clean negative control.
HPA tissue expression evidence for PRKACB
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | lymphoid tissue | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | High | Protein (IHC) | HPA → |
| Pancreas | exocrine glandular cells | High | Protein (IHC) | HPA → |
| Spleen | cells in white pulp | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bronchus | respiratory epithelial cells | Low | Protein (IHC) | HPA → |
| Cervix | glandular cells | Low | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced PRKACB Western Blot Tips
Deeper troubleshooting and optimisation questions for PRKACB, answered from its protein features.
How should PRKACB band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
How could PRKACB isoforms affect the blot?
Isoforms · Ten isoforms are listed. Several replace or remove N-terminal residues, while isoform 10 lacks residues 79–111 and isoform 8 lacks residues 258–351. Check which isoforms your antibody recognizes before assigning multiple bands.
Yes. Isoforms 3 and 4 lack canonical residues 1–12 and 1–13, respectively; several others replace part of the N terminus. Compare the antibody epitope with each relevant isoform sequence before interpreting an absent band.
Yes. Isoforms 3 and 4 lack canonical residues 1–12 and 1–13, respectively; several others replace part of the N terminus. Compare the antibody epitope with each relevant isoform sequence before interpreting an absent band.
Which PRKACB modifications matter when interpreting bands?
PTM · The supplied UniProt features list deamidated Asn3 and phosphorylation at Ser11, Tyr69, Ser140, Thr198, Ser322, Tyr331, and Ser339. These are canonical UniProt coordinates; paper or antibody numbering may differ. Confirm the detected species before attributing a band shift to any modification.
Does cAMP induction imply more PRKACB protein?
Induction · The supplied holoenzyme feature says cAMP causes dissociation of inactive regulatory–catalytic complexes. The location feature notes nuclear translocation of the monomeric catalytic subunit. Neither establishes increased PRKACB abundance; compare total protein and the relevant cellular fractions separately.
How should transfer be checked for PRKACB?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05366-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should PRKACB be quantified across cellular fractions?
Quantitation · PRKACB is listed in the cytoplasm, at membranes, and in the nucleus; the inactive holoenzyme is cytoplasmic, while the monomeric catalytic subunit translocates to the nucleus. Quantify comparable fractions consistently and distinguish a distribution change from a change in total PRKACB.
Should PRKACB appear exactly at 40.6 kDa?
Interpretation · 40.6 kDa is the predicted mass for the supplied canonical sequence. No observed band position is provided. Alternative sequences and modifications may affect migration, but their presence alone does not establish a visible shift or explain a measured mass difference.
First compare them with the canonical 40.6 kDa prediction and the listed isoform sequence changes, especially the deletions in isoforms 8 and 10. Check antibody epitope coverage and whether the band tracks PRKACB. The supplied features alone cannot identify an unexpected band.
First compare them with the canonical 40.6 kDa prediction and the listed isoform sequence changes, especially the deletions in isoforms 8 and 10. Check antibody epitope coverage and whether the band tracks PRKACB. The supplied features alone cannot identify an unexpected band.
Boster reagents
PRKACB Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-PKA Beta cat PRKACB Antibody
Cat # A05366-1
The catalog reports A05366-1 as an anti-PRKACB antibody with reported Human, Mouse, and Rat reactivity. Its WB image uses extracts from various cell lines at 1:500 and 25 µg per lane. The caption does not identify the cell lines or their species.
Which to pick: A05366-1 is the only listed antibody and has a WB image. Consider it if its reported reactivity matches your sample, while noting that the image shows unspecified cell-line extracts rather than validation across all listed species.
Source: BosterBio PRKACB gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P22694.
- Human Protein Atlas. PRKACB tissue expression.
- PMC12902190 — target-verified WB comparison
- PMC3700962 — target-verified WB comparison
- PMC7608482 — target-verified WB comparison