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PRKACB / cAMP-dependent protein kinase catalytic subunit beta · IHC design guide
Design Immunohistochemistry for PRKACB
Plan PRKACB chromogenic IHC around the general cytoplasmic tissue pattern (HPA tissue IHC). Cerebral cortex neurons and appendix lymphoid tissue show high staining (HPA tissue IHC); titrate catalog antibody A05366-1 within 1:50–1:200 (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PRKACB Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Splice or transcript discrepancy may affect staining (HPA tissue IHC) | |
| Regulation | Tissue-specific isoform expression (UniProt) | |
| Isoform / epitope | 10 isoforms; epitope coverage needs validation (UniProt) |
Section 1
Recommended PRKACB IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published PRKACB IHC methods for gastric, osteosarcoma, testicular and esophageal tissue (PMC12006100; PMC10244582; PMC11559180; PMC12831332).
| Sample | Paraffin-embedded rat brain tissue; fixative not specified (datasheet A05366-1) |
| Fixation | Image fixative and duration unreported (datasheet A05366-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-PRKACB, 1:50-1:200 (datasheet A05366-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PRKACB-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval rule); compare the reported citrate conditions if needed (PMC12006100; PMC11559180).
Section 2
What Is the Expected PRKACB Staining Pattern?
PRKACB should appear mainly cytoplasmic in paraffin-section IHC, with high staining reported in cerebral-cortex neurons, pancreatic exocrine cells and selected lymphoid populations (HPA: general cytoplasmic expression; High in these cells). Nuclear localisation is biologically possible after catalytic-subunit translocation (UniProt P22694: subcellular location). PRKACB has no transmembrane segment, so a membrane-only pattern needs scrutiny (UniProt P22694: topology). HPA rates the tissue IHC profile Approved, with a splice/transcript-discrepancy caution (HPA: tissue reliability).
| Clear cytoplasmic stain in cerebral-cortex neurons, pancreatic exocrine cells or splenic white-pulp cells (HPA: High in these cells). | This fits the reported tissue pattern (HPA: tissue IHC). Compare the intended cell population with surrounding cells: a positive tissue need not stain uniformly because HPA reports intensity by cell population (HPA: tissue IHC). |
| Predominantly nuclear or sharply membrane-only stain, with little cytoplasmic signal (HPA: general cytoplasmic expression). | Nuclear signal can reflect catalytic-subunit translocation, but nuclear dominance alone does not establish pathway activation (UniProt P22694: subcellular location). A membrane-only pattern is less consistent with the reported cytoplasmic profile and no-transmembrane topology; check staining controls (HPA: tissue IHC; UniProt P22694: topology; general IHC practice). |
| Strong stain mainly in bronchial respiratory epithelium or liver cholangiocytes, while expected positive cells are weak (HPA: Low in these cells). | That reversal conflicts with the reported cell-level pattern (HPA: tissue IHC). Consider off-target binding or detection-system activity, then compare an appropriate negative control and a known-positive section (general IHC practice). Low means some signal may be genuine; it does not mean absent expression (HPA: Low). |
| Diffuse color over stroma, lumens and cells, obscuring cell boundaries (general IHC practice). | This is not the reported cell-associated cytoplasmic pattern (HPA: tissue IHC). Background may arise from nonspecific reagent binding, incomplete blocking or endogenous detection activity; use control sections to locate its source (general IHC practice). |
| No discernible stain in cerebral-cortex neurons or pancreatic exocrine cells (HPA: High in these cells). | Treat the run as unresolved before calling the specimen negative: verify tissue preservation, retrieval, antibody and detection steps against the catalog antibody’s IHC-P instructions and controls (general IHC practice). HPA's Approved status has a splice/transcript-discrepancy caution (HPA: tissue reliability). |
💡Expected PRKACB appearanceA convincing positive is cell-associated, predominantly cytoplasmic color in HPA-high populations such as cerebral-cortex neurons or pancreatic exocrine cells; widespread acellular haze or staining chiefly in HPA-low cells calls for control-based review (HPA: tissue IHC; general IHC practice).
| Cell population and isoforms (HPA: tissue IHC; UniProt P22694: tissue specificity). | HPA reports high staining in several neuronal, glandular and lymphoid populations; UniProt reports tissue-dependent isoforms. A tissue name alone cannot predict every cell's signal (HPA: tissue IHC; UniProt P22694: tissue specificity). |
| Compartment and signaling state (UniProt P22694: subcellular location). | Inactive holoenzyme is cytoplasmic; free catalytic subunit can enter the nucleus. Interpret nuclear signal alongside the dominant cytoplasmic IHC pattern and controls (UniProt P22694: subcellular location; HPA: tissue IHC; general IHC practice). |
| Antibody validation scope (HPA: antibodies; tissue reliability). | HPA029754 and CAB010363 are IHC Approved, but the tissue profile carries a splice/transcript-discrepancy caution. Approved status does not identify which isoform an individual stain detects (HPA: antibodies; tissue reliability; UniProt P22694: isoforms). |
| Antigen retrieval and detection (general IHC practice). | For paraffin IHC, follow the catalog antibody’s IHC-P retrieval and detection instructions and include run controls. These sources give no PRKACB-specific fixation sensitivity or retrieval effect (general IHC practice; HPA: tissue IHC; UniProt P22694: record). |
| IF/ICC Q: Where should PRKACB appear? (HPA: subcellular ICC-IF). | A: Mainly cytosol; microtubules are an additional approved location. Cilium, basal-body and cytokinetic-bridge calls are uncertain, and HPA cautions that the summary draws on antibodies targeting multiple genes (HPA: subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cells are unstained (HPA: High in cerebral-cortex neurons and pancreatic exocrine cells). | Possible failure in retrieval, primary-antibody or chromogenic detection steps (general IHC practice). | Repeat with a known-positive section and the catalog antibody’s IHC-P instructions; check each run control before interpreting the specimen (general IHC practice). |
| Strong signal appears in a no-primary control (general IHC practice). | Endogenous detection activity or nonspecific detection-reagent binding may contribute (general IHC practice). | Review the detection system and its appropriate blocking and control steps; reassess specimen signal only after control background is resolved (general IHC practice). |
| Color covers the section diffusely (general IHC practice). | Possible nonspecific binding, inadequate blocking or excessive detection background (general IHC practice). | Compare no-primary and known-positive controls; check reagent concentrations, wash steps and blocking against the established IHC workflow (general IHC practice). |
| Nuclei dominate while cytoplasm is faint (HPA: general cytoplasmic expression). | Nuclear entry is possible, but staining alone cannot establish activation; technical background is also possible (UniProt P22694: subcellular location; general IHC practice). | Check controls and whether nuclear staining is confined to relevant cells; report the observed compartment without inferring signaling state (general IHC practice; UniProt P22694: subcellular location). |
| HPA-low cells stain more intensely than expected high cells (HPA: tissue IHC). | Possible off-target staining, detection background or differences among cell populations; HPA Low is not absence (general IHC practice; HPA: Low). | Compare matched positive and negative controls, then score the named cell populations separately instead of labeling the whole tissue positive (general IHC practice; HPA: tissue IHC). |
| IHC and IF/ICC show different minor compartments (HPA: tissue IHC; subcellular ICC-IF). | The assays report different observations; HPA's ICC-IF summary cautions that antibodies target proteins from multiple genes (HPA: subcellular ICC-IF). | Prioritize the IHC cell-level cytoplasmic profile when scoring paraffin sections; describe any additional compartment with its assay and HPA confidence level (HPA: tissue IHC; subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing PRKACB staining
A05366-1 · IHCImmunohistochemistry of paraffin-embedded rat brain using PKA C-beta antibody at dilution of 1:100 .Perform microwave antigen retrieval with
A05366-1 · IHCImmunohistochemistry of paraffin-embedded human appendix using PKA C-beta antibody at dilution of 1:100 .Perform microwave antigen retrieval
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000142875-PRKACB/tissue/cerebral+cortex#imid_7258528
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000142875-PRKACB/tissue/colon#imid_7266294
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000142875-PRKACB/tissue/liver#imid_7266295
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000142875-PRKACB/tissue/pancreas#imid_7266254
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA071185; green = PRKACB, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA071185; green = PRKACB, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PRKACB IHC & IF
🧪Run cerebral cortex first and expect staining in neuronal cells (HPA: High in cerebral cortex neuronal cells). HPA detects PRKACB in all 45 scored tissues, so use no-primary and isotype controls for the negative; any unstained cells on the positive slide should show only background, without treating them as a validated PRKACB-negative cell population (HPA: no negative tissue rows; standard IHC practice).
Positive control tissue: Appendix (Lymphoid tissue, HPA High)
Negative control tissue: None in HPA: PRKACB is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PRKACB in MCF-7, PC-3, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a host-matched nonimmune IgG control for a polyclonal antibody or a matched isotype control for a monoclonal antibody; use PRKACB-knockout material as a biological negative if available (standard IHC practice). Quench endogenous peroxidase for HRP detection and check cerebral cortex autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A05366-1 rat-brain paraffin-section caption does not state the fixative (selected-SKU tissue-IHC caption). That caption uses microwave retrieval in 10 mM PBS at pH 7.2 and a 1:100 dilution; whether retrieval is required needs a paired comparison (selected-SKU tissue-IHC caption; standard IHC practice). The supplied evidence does not establish that frozen sections or IF are easier; IF in brain may be complicated by tissue autofluorescence (standard IF practice).
HPA tissue IHC evidence for PRKACB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Caution, Splice and/or transcript discrepancy exists.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| 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 → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PRKACB is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PRKACB IHC Tips
Troubleshoot PRKACB chromogenic IHC by checking retrieval, tissue handling, cellular localisation and scoring against the reported expression patterns.
Which retrieval conditions should I try when PRKACB staining is weak?
Start IHC-P with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval setting: nuclear-antigen rule). If staining remains weak, compare a section retrieved by microwave in 10 mM PBS, pH 7.2, as reported for the catalog antibody on paraffin-embedded rat brain (caption: A05366-1). Keep antibody dilution and detection conditions matched across sections; the reported image used 1:100 antibody dilution (caption: A05366-1). Score preserved morphology alongside specific cytoplasmic signal, since excessive heat can damage sections and obscure cellular boundaries (general IHC practice; HPA: general cytoplasmic expression).
How should I troubleshoot variable PRKACB staining after fixation?
PRKACB-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence (caption: A05366-1, fixative not stated). The catalog image documents paraffin-embedded rat brain but does not establish how that tissue was fixed (caption: A05366-1). For IHC troubleshooting, record fixation conditions for each specimen and compare sections processed with the same retrieval, antibody dilution and chromogenic detection conditions (general IHC practice). Examine morphology and staining together: poorly preserved areas can complicate interpretation, while a consistent signal in intact cells is more useful for comparing specimens (general IHC practice).
Should PRKACB staining be cytoplasmic, nuclear or membranous?
Expect predominantly cytoplasmic staining in tissue IHC, consistent with the reported general cytoplasmic expression pattern (HPA: tissue IHC profile). Nuclear signal can be biologically plausible because the free catalytic subunit translocates to the nucleus, whereas inactive holoenzyme resides in the cytoplasm (UniProt P22694: subcellular location and subunit). UniProt also lists membrane localisation but reports no transmembrane segment, so do not require a continuous cell-surface rim to call a section positive (UniProt P22694: location and topology). Compare nuclear and cytoplasmic staining within intact cells and across matched conditions before attributing a shift to cAMP signalling (UniProt P22694: subunit; general IHC practice).
Could isoforms or epitope accessibility explain tissue-dependent staining?
PRKACB has 10 listed isoforms, and their reported tissue distributions differ (UniProt P22694: isoforms and tissue specificity). Isoform 1 is most abundant in brain, isoform 2 predominates in thymus, spleen and kidney, and isoforms 3 and 4 are reported only in brain (UniProt P22694: tissue specificity). Before comparing tissues, check whether the antibody's documented epitope is shared among the isoforms being assessed; the supplied caption does not identify that epitope (caption: A05366-1; general IHC practice). The protein also has reported modified residues, including phosphothreonine at position 198, but their effect on this antibody's IHC staining is unknown (UniProt P22694: modified residues; caption: A05366-1).
How can I check an IHC localisation result using IF multiplexing?
Use IF as a separate check of the IHC pattern, pairing PRKACB with a marker of the expected cell type and including single-label controls (general IF practice). For example, neuronal cells show high PRKACB tissue staining in cerebral cortex, so the companion marker should identify neuronal cells if that is the population under study (HPA: cerebral cortex, neuronal cells High). Choose fluorophores and imaging channels after checking tissue autofluorescence, and assess bleed-through with single-label controls (general IF practice). For intracellular PRKACB epitopes, optimise permeabilisation to provide cytosolic or nuclear access as appropriate; the protein has no transmembrane segment, and its reported locations include cytoplasm and nucleus (UniProt P22694: topology and subcellular location; general IF practice).
What should I change when chromogenic PRKACB staining looks diffuse?
First compare the section with a no-primary control and inspect whether color follows tissue folds, cut edges or damaged areas (general IHC practice). Check the peroxidase block and chromogen development because endogenous peroxidase activity and prolonged development can add nonspecific color in chromogenic IHC (general IHC practice). Then adjust blocking, washing and antibody exposure systematically while keeping retrieval constant; the catalog rat-brain image reports a 1:100 dilution for its stated conditions (caption: A05366-1; general IHC practice). Retain a signal only when it resolves into intact cells and fits the reported general cytoplasmic tissue pattern (HPA: tissue IHC profile; general IHC practice).
How should I score PRKACB staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since PRKACB tissue IHC is generally cytoplasmic but nuclear translocation is reported (HPA: tissue IHC profile; UniProt P22694: subcellular location). Within a consistent region, report the percentage of positive cells and an intensity-based H-score, with the scoring thresholds fixed across slides (general IHC practice). Normalise cell counts to the number of eligible intact cells, or report positive-cell density per mm² of analysable tissue when cell segmentation is unreliable (general IHC practice). Score cytoplasmic and nuclear signals separately, and exclude folds, necrotic regions and poorly preserved tissue using the same rules for every specimen (general IHC practice).
How can I distinguish genuine PRKACB staining from artefact?
A credible IHC result localises to intact cells in a plausible compartment: general cytoplasmic staining is reported, and nuclear localisation is biologically possible (HPA: tissue IHC profile; UniProt P22694: subcellular location). Check the expected cell population as well as intensity; neuronal cells in cerebral cortex and cells in splenic white pulp are reported as High (HPA: cerebral cortex, neuronal cells High; spleen, white-pulp cells High). Staining confined to edges, folds or necrotic tissue warrants suspicion, particularly if it also appears in the no-primary control (general IHC practice). Diffuse chromogen in that control calls for checking endogenous enzyme blocking and detection conditions before interpreting PRKACB expression (general IHC practice).
Boster reagents
Best PRKACB / cAMP-dependent protein kinase catalytic subunit beta IHC Antibodies
The catalog lists PRKACB antibodies for IHC and IF/ICC in human, mouse and rat; IHC images show paraffin-embedded rat brain and human appendix (catalog: applications, reactivity and A05366-1 IHC captions).
Real IHC data
Anti-PKA Beta cat PRKACB Antibody
Cat # A05366-1
A05366-1 lists IHC and IF/ICC for human, mouse and rat, with IHC images of paraffin-embedded rat brain and human appendix (catalog: A05366-1 applications, reactivity and IHC captions). A05366-3 lists IHC and IF/ICC for the same species, but has no IHC or IF image in the payload (catalog: A05366-3 applications, reactivity and image fields).
Which to pick: For tissue IHC, choose A05366-1: its own captions document paraffin-embedded rat brain and human appendix at 1:100 after microwave retrieval in 10 mM PBS, pH 7.2; the fixative is unreported (catalog: A05366-1 IHC captions). For IF/ICC, A05366-1 lists both applications at 1:50–1:200; A05366-3 is a listed polyclonal option at 1:50, with no IF image reported for either SKU (catalog: applications, dilution fields, A05366-3 dilution_raw and image fields). Both SKUs list human, mouse and rat reactivity, but the documented tissue IHC examples belong to A05366-1 (catalog: reactivity and A05366-1 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P22694 (KAPCB_HUMAN, cAMP-dependent protein kinase catalytic subunit beta).
- Human Protein Atlas. PRKACB tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PRKACB subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the microtubules, cytokinetic bridge, primary cilium and basal body. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. PRKACB antibody validation summary (3 antibodies).
- Alpha-estradiol and (R)-(-)-ibuprofen inhibit gastric cancer progression via GLI1 G-quadruplex. Frontiers in pharmacology 2025 — PMC12006100.
- A glycometabolic gene signature associating with immune infiltration and chemosensitivity and predicting the prognosis of patients with osteosarcoma. Frontiers in medicine 2023 — PMC10244582.
- Prediction model of male reproductive function damage caused by CHOP chemotherapy regimen for non-Hodgkin's lymphoma. BMC cancer 2024 — PMC11559180.
- Porphyromonas gingivalis extracellular vesicles promotes tumor metastasis in esophageal squamous cell carcinoma by inducing PRKACB/JNK/ NFATC2 axis. Journal of nanobiotechnology 2025 — PMC12831332.
- PubMed PMID:2342480 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PRKACB (cAMP-dependent protein kinase catalytic subunit beta). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A05366-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat brain tissue; fixative not specified (datasheet A05366-1); 2. Fixation: Image fixative and duration unreported (datasheet A05366-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-PRKACB, 1:50-1:200 (datasheet A05366-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Splice or transcript discrepancy may affect staining (HPA tissue IHC); Isoform / epitope: 10 isoforms; epitope coverage needs validation (UniProt); Regulation: Tissue-specific isoform expression (UniProt). Evidence: UniProt P22694, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A05366-1, PMC12006100, PMC10244582. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PRKACB/immunohistochemistry-ihc-guide.