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PKIB / cAMP-dependent protein kinase inhibitor beta · IHC design guide
Design Immunohistochemistry for PKIB
Plan chromogenic PKIB IHC in paraffin sections using the tissue staining profile (HPA tissue IHC) and the IHC-validated antibody’s 2–5 μg/ml range (datasheet A12582-3). Compare cytoplasmic staining across cell types and account for possible macrophage background (HPA tissue IHC; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PKIB 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 | Cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic staining; high in lung macrophages (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A12582-3) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Spleen |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A12582-3) | |
| Caveat | Lung macrophages may show endogenous peroxidase background (HPA tissue IHC; standard IHC practice) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unknown; no TM segment (UniProt) |
Section 1
Recommended PKIB IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A12582-3); the published placental PKIB IHC protocol uses citrate pH 6.0 retrieval (PMC7782383).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet A12582-3) |
| Fixation | Image fixative and duration unreported (datasheet A12582-3); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A12582-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A12582-3) |
| Primary antibody | Rabbit anti-PKIB, 2-5 μg/ml (datasheet A12582-3) |
| Primary incubation | Overnight at 4 °C (datasheet A12582-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A12582-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PKIB-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet A12582-3). Citrate pH 6.0 is a published alternative for placental sections (PMC7782383).
Section 2
What Is the Expected PKIB Staining Pattern?
PKIB should appear predominantly in the cytoplasm of stained cells (HPA: ubiquitous cytoplasmic IHC profile). Strong examples include kidney tubular cells, lung macrophages, and breast myoepithelial cells (HPA: High in each). The tissue IHC assessment is Approved, with medium consistency between staining and RNA data (HPA). PKIB has no annotated transmembrane segment, while UniProt assigns no subcellular location (UniProt Q9C010 topology and subcellular record).
| Cytoplasmic staining is strong in kidney tubular cells or lung macrophages, with recognizable tissue architecture. | This matches two documented high-staining cell populations (HPA: High in kidney tubular cells and lung macrophages). Judge the named cells and their cytoplasm; staining elsewhere in the section alone does not establish the expected result (HPA: tissue IHC profile). |
| A crisp nuclear-only or membrane-rim pattern dominates the cells being scored. | Treat the compartment mismatch as a reason to investigate artefact or antibody specificity (HPA: cytoplasmic IHC profile; UniProt Q9C010: no annotated transmembrane segment). UniProt provides no independent PKIB subcellular annotation, so the HPA pattern is the available location reference (UniProt Q9C010; HPA). |
| Strong signal appears mainly in cells outside the documented high-staining population within a reference tissue. | For example, a kidney section should be evaluated in tubular cells, and a lung section in macrophages (HPA: High in those cells). Staining of other cells warrants a check for cross-reactivity or endogenous detection activity; it cannot by itself be assigned to PKIB (general IHC practice). |
| Color spreads across tissue, extracellular spaces, or areas without discernible cellular borders. | A diffuse field is hard to reconcile with interpretable cytoplasmic staining (HPA: cytoplasmic IHC profile). Compare a no-primary control and inspect washing, blocking, and detection steps to identify background from the staining workflow (general IHC practice). |
| A kidney section shows no signal in tubular cells, or a lung section shows none in macrophages. | These are documented high-staining populations, so a blank result calls for a run-level check before concluding PKIB is absent (HPA: High in kidney tubular cells and lung macrophages). Verify tissue identity, reagent performance, retrieval conditions, and detection controls (general IHC practice). |
💡Expected PKIB appearanceCall a result positive when staining is predominantly cytoplasmic and clearly visible in a documented high-staining population, such as kidney tubular cells or lung macrophages (HPA: cytoplasmic profile; High in those cells); nuclear-only, membrane-rim, or cell-free diffuse color calls for investigation rather than a PKIB-positive score (HPA: cytoplasmic profile; general IHC practice).
| Cell population and tissue choice | HPA reports High staining in adrenal gland and gallbladder glandular cells, breast myoepithelial cells, caudate neurons, cardiomyocytes, kidney tubular cells, lung macrophages, and nasopharyngeal respiratory epithelium (HPA: tissue IHC). Select a documented cell population when judging whether a section behaved as expected. |
| Reference range and validation strength | Prostate glandular cells, ovarian stromal cells, soft-tissue fibroblasts, and skin keratinocytes are Low; spleen red-pulp cells are Not detected (HPA: tissue IHC). These observations describe particular cells, not whole-organ negative controls. HPA rates tissue IHC Approved and reports medium staining–RNA consistency (HPA). |
| Assay and location evidence | The HPA location statement comes from tissue IHC (HPA: ubiquitous cytoplasmic expression). The listed antibody HPA030156 is IHC Approved, while the supplied ICC-IF record has no main location or cell-line images (HPA: antibody and subcellular records). For IF/ICC, use the cytoplasmic IHC pattern as a comparison, not as an independently validated IF pattern. |
| Isoforms and epitope interpretation | UniProt lists PKIB isoforms 1 and 2 but supplies no antibody epitope in this record (UniProt Q9C010: alternative splicing). Whether a given antibody detects both isoforms cannot be inferred here; consult its epitope documentation before interpreting an isoform-specific difference (general IHC practice). |
| Topology and processing | PKIB is a 78-residue chain with no annotated signal peptide, propeptide, transmembrane segment, glycosylation site, or modified residue (UniProt Q9C010). These annotations support caution with a membrane-only interpretation; they do not predict staining intensity or an antigen-retrieval requirement. |
| Paraffin-section workflow | No PKIB-specific fixation sensitivity, retrieval condition, or dilution is supplied (HPA and UniProt records provided). If signal is poor, compare retrieval and detection conditions using the same documented high-staining tissue and appropriate controls (general IHC practice); do not assign a PKIB-specific fixation effect from these sources. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in kidney tubular cells or lung macrophages | A high-staining reference population failed to appear (HPA: High in both); tissue identity or an IHC step may need checking (general IHC practice). | Confirm the expected cells are present, then check primary-antibody application, retrieval, chromogen, and control-slide performance (general IHC practice). Interpret the run only after a documented reference population stains. |
| Nuclei dominate while cytoplasm remains pale | The observed compartment conflicts with HPA's cytoplasmic tissue profile (HPA: ubiquitous cytoplasmic expression). The source record does not establish a nuclear PKIB pattern (HPA; UniProt Q9C010: location unannotated). | Compare with a no-primary control and a documented high-staining tissue; review morphology and staining conditions before scoring the nuclear color as PKIB (general IHC practice). |
| Strong color marks unexpected cells in a reference section | Cell identity may have been misread, or cross-reactivity or endogenous detection activity may contribute (general IHC practice). HPA's High calls refer to named populations, such as kidney tubules and lung macrophages (HPA: tissue IHC). | Identify the stained cells morphologically; compare a no-primary control and, if available, an independent antibody before attributing the unexpected population to PKIB (general IHC practice). |
| Diffuse color obscures cell boundaries | Nonspecific background from blocking, washing, or detection can make cytoplasmic signal uninterpretable (general IHC practice; HPA: cytoplasmic profile). | Inspect the no-primary control and reagent-only background; review blocking, wash, and chromogen steps, then score only cells with resolved cytoplasmic staining (general IHC practice). |
| A Low or Not detected reference looks strongly positive | The result conflicts with the reported level for the specific cells: prostate glandular cells are Low and spleen red-pulp cells are Not detected (HPA: tissue IHC). These HPA levels are observations, not absolute exclusion of every signal. | Verify the cell population and compare the same run with a documented High population and background controls; investigate excess signal before treating it as a PKIB expression difference (HPA: tissue IHC; general IHC practice). |
| An IF/ICC image is being used to settle an IHC compartment dispute | The supplied HPA ICC-IF record has no main location or cell-line images, whereas HPA's cytoplasmic statement is from tissue IHC (HPA: subcellular and tissue records). | Base this paraffin-section interpretation on tissue morphology and the documented IHC pattern (HPA: tissue IHC). Treat any IF/ICC result as separate evidence requiring its own validation (general IHC practice). |
Experiment evidence
IHC & IF images showing PKIB staining
A12582-3 · IHCIHC analysis of PKIB using anti-PKIB antibody (A12582-3). PKIB was detected in a paraffin-embedded section of human placenta tissue. Heat me
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000135549-PKIB/tissue/cerebral+cortex#imid_7520384
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000135549-PKIB/tissue/colon#imid_7520318
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000135549-PKIB/tissue/liver#imid_7520319
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000135549-PKIB/tissue/kidney#imid_7520320
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PKIB IHC & IF
🧪Run breast first and look for staining in myoepithelial cells (HPA: High in breast myoepithelial cells); use spleen red pulp as the negative tissue (HPA: Not detected in spleen red-pulp cells). On the breast slide, compare myoepithelial staining with adjacent luminal cells: they should show no specific chromogen signal to serve as internal negative cells, but verify this on the slide rather than assuming they are PKIB-negative from HPA.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Spleen (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PKIB; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control, a rabbit IgG isotype control matched to the catalog antibody’s clonality, and a biological negative such as PKIB-knockout tissue or a validated peptide-block control (datasheet: rabbit anti-PKIB antibody; standard IHC practice). Quench endogenous peroxidase before HRP/DAB detection in breast sections (datasheet: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A12582-3 paraffin-section caption does not state the fixative (datasheet: A12582-3 tissue-IHC caption). The caption reports heat-mediated retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required or whether another method works (datasheet: A12582-3 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; in breast sections, the close position of myoepithelial and luminal cells makes cell assignment a scoring concern (HPA: High in breast myoepithelial cells; standard breast histology).
HPA tissue IHC evidence for PKIB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Myoepithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PKIB IHC Tips
Troubleshoot PKIB staining in paraffin sections by checking retrieval, cell type, cytoplasmic pattern, and controls before comparing signal intensity.
What retrieval should I use when PKIB staining is weak in paraffin sections?
Use heat-mediated antigen retrieval in EDTA at pH 8.0 for PKIB IHC (datasheet A12582-3). The selected paraffin-section image used that retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C (caption A12582-3). If staining is weak, check that sections experienced consistent heating, cooling, and buffer coverage before changing primary-antibody concentration (standard IHC practice). Compare any retrieval adjustment on adjacent sections with the same detection conditions and a known positive cell population, such as lung macrophages, so a stronger DAB signal can be distinguished from broader background (HPA: High in lung macrophages).
Could fixation explain weak or uneven PKIB staining?
Target-specific PKIB sensitivity to fixation is unknown: the selected paraffin-section caption does not report a fixative (caption A12582-3). Record the specimen’s actual fixative and fixation interval, then compare sections processed under matched conditions with the established EDTA pH 8.0 retrieval (datasheet A12582-3; standard IHC practice). Evaluate morphology and staining together, since poor preservation or uneven reagent access can complicate interpretation of any chromogenic stain (standard IHC practice). Do not assign a fixation effect to PKIB from its lack of an annotated transmembrane segment or modified residues; those annotations do not test fixation sensitivity (UniProt Q9C010 topology and modified-residue record).
Which staining compartment and cell populations support a PKIB IHC result?
Assess PKIB primarily as cytoplasmic staining, while recognizing that UniProt supplies no subcellular-location annotation for this protein (HPA: ubiquitous cytoplasmic expression; UniProt Q9C010 subcellular record). For a cell-specific check, HPA reports High staining in kidney tubule cells and lung macrophages, whereas spleen red-pulp cells are Not detected (HPA tissue IHC). Compare the fraction and intensity of cytoplasmic signal within identified cells on sections developed for the same 30 minutes of secondary-antibody incubation when following the selected caption (caption A12582-3). Predominantly nuclear or extracellular DAB should prompt review of morphology, background controls, and antibody specificity before being scored as PKIB (HPA: cytoplasmic profile; standard IHC practice).
Can this IHC assay distinguish PKIB isoforms or predict epitope masking?
PKIB has 2 annotated isoforms, but the supplied antibody caption does not map its epitope or establish isoform-specific detection (UniProt Q9C010 isoform record; caption A12582-3). Treat a positive DAB signal as antibody-associated PKIB staining rather than assigning it to isoform 1 or 2 without separate validation (UniProt Q9C010 isoform record; standard IHC practice). The record lists no annotated glycosylation sites, modified residues, or domains, which provides no direct test of epitope accessibility after tissue processing (UniProt Q9C010 annotations). If signal varies between specimens, hold EDTA pH 8.0 retrieval and detection conditions constant while assessing tissue preservation and cell identity (datasheet A12582-3; standard IHC practice).
How should I assess PKIB by IF alongside a cell-type marker?
Treat IF as a separate assay requiring its own validation; the selected antibody evidence documents paraffin-section chromogenic IHC, and HPA lists no cell lines with PKIB ICC/IF images (caption A12582-3; HPA subcellular record). For multiplexing, pair PKIB with a validated macrophage marker when examining lung macrophages, a High-staining population in tissue IHC (HPA: High in lung macrophages; standard IF practice). Choose a spectrally separated, preferably far-red PKIB channel after checking tissue autofluorescence in unstained controls (standard IF practice). Because the reported tissue pattern is cytoplasmic and PKIB lacks an annotated transmembrane segment, assess whether permeabilisation permits access to the antibody’s unknown epitope; do not assume its orientation (HPA: cytoplasmic profile; UniProt Q9C010 topology; standard IF practice).
How can I separate PKIB staining from diffuse DAB background?
Inspect a no-primary control and a matched section for diffuse DAB, tissue pigment, and signal near damaged edges before interpreting weak cytoplasmic staining (standard IHC practice; HPA: cytoplasmic profile). The selected caption used 10% goat serum for blocking and a peroxidase-conjugated secondary for 30 minutes at 37°C (caption A12582-3). An endogenous-peroxidase block and careful DAB development are general chromogenic IHC controls, not evidence that PKIB itself creates background (standard IHC practice). If background persists, compare reagent omissions and shorter chromogen development on adjacent sections while checking whether expected lung macrophages retain cell-associated signal (HPA: High in lung macrophages; standard IHC practice).
What is a defensible way to quantify PKIB IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysis region and cell population before scoring, using matched retrieval in EDTA at pH 8.0 and matched imaging conditions across sections (datasheet A12582-3; standard IHC practice). Report the percentage of positive viable cells and an H-score from 0–300, calculated by weighting the percentages at staining intensities 0–3 (standard IHC practice). If counting positive cells per mm², also report viable tissue area and normalize cell-based comparisons to the number of eligible, identified cells (standard IHC practice). Score cytoplasmic DAB within the chosen cell type and keep edge artefact, necrosis, and tissue pigment outside the measurement mask (HPA: cytoplasmic profile; standard IHC practice).
When should an apparent PKIB-positive IHC result be questioned?
A convincing result shows cell-associated cytoplasmic staining in morphologically intact cells, consistent with the reported tissue-IHC profile (HPA: ubiquitous cytoplasmic expression; standard IHC practice). Check cell identity: HPA reports High staining in lung macrophages but Not detected in spleen red-pulp cells, so a universal strong signal across those populations warrants review (HPA tissue IHC). Discount staining confined to section edges, necrotic areas, or structures also colored in no-primary controls, and check an endogenous-peroxidase control when DAB is used (standard IHC practice). The selected image demonstrates paraffin-section staining after EDTA pH 8.0 retrieval; it does not establish specificity in every specimen or report the fixative (caption A12582-3).
Boster reagents
Best PKIB / cAMP-dependent protein kinase inhibitor beta IHC Antibodies
The IHC-validated anti-PKIB antibody has real chromogenic IHC data from a human paraffin-embedded placenta section (A12582-3 image caption); no IF figure is provided (catalog).
Real IHC data
Anti-PKIB Antibody
Cat # A12582-3
A12582-3 will render with an IHC figure showing human paraffin-embedded placenta tissue (A12582-3 image caption). Its listed applications are IHC and ELISA, and its listed reactivity is human (catalog).
Which to pick: Choose A12582-3, a polyclonal antibody, for human paraffin-section IHC (catalog; A12582-3 image caption). Its caption reports a paraffin-embedded section but does not report the fixative (A12582-3 image caption). No SKU in the payload is listed for IF/ICC or cross-species use (catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9C010 (IPKB_HUMAN, cAMP-dependent protein kinase inhibitor beta).
- Human Protein Atlas. PKIB tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PKIB subcellular location (ICC-IF): Highest expression in RPTEC/TERT1: 173.1 nTPM.
- Human Protein Atlas. PKIB antibody validation summary (1 antibodies).
- PKIB facilitates bladder cancer proliferation and metastasis through mediation of HSP27 phosphorylation by PKA. Cell death & disease 2025 — PMC12219054.
- Downregulation of cAMP-Dependent Protein Kinase Inhibitor-b Promotes Preeclampsia by Decreasing Phosphorylated Akt. Reproductive sciences (Thousand Oaks, Calif.) 2021 — PMC7782383.
- PubMed PMID:10880337 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PKIB (cAMP-dependent protein kinase inhibitor beta). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A12582-3). Validated IHC antibody: A12582-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Spleen. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human placenta tissue; fixative not specified (datasheet A12582-3); 2. Fixation: Image fixative and duration unreported (datasheet A12582-3); 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 retrieval: EDTA pH 8.0 (datasheet A12582-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A12582-3); 8. Primary antibody: Rabbit anti-PKIB, 2-5 μg/ml (datasheet A12582-3); 9. Primary incubation: Overnight at 4 °C (datasheet A12582-3). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A12582-3); Caveat: Lung macrophages may show endogenous peroxidase background (HPA tissue IHC; standard IHC practice); Isoform / epitope: 2 isoforms; epitope differences unknown; no TM segment (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt Q9C010, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in RPTEC/TERT1: 173.1 nTPM, Datasheet A12582-3, PMC7782383. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PKIB/immunohistochemistry-ihc-guide.