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PRKAB2 / 5'-AMP-activated protein kinase subunit beta-2 · IHC design guide
Design Immunohistochemistry for PRKAB2
Plan PRKAB2 paraffin-section IHC around cytoplasmic staining in respiratory epithelial cells, glandular cells and hepatocytes (HPA tissue IHC). Use consistent fixation, score cytoplasmic signal and start with the IHC-validated antibody at 0.5–1 μg/mL (datasheet PB9739).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PRKAB2 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 | Cytoplasm in respiratory epithelium, glands and hepatocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9739) | |
| Positive control | Bronchus+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 | Staining and RNA show only medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; confirm epitope coverage (UniProt) |
Section 1
Recommended PRKAB2 IHC & IF Protocols
The catalog antibody protocol (datasheet: PB9739) and one published PRKAB2 IHC protocol (PMC13232994) provide starting conditions for paraffin sections.
| Sample | Paraffin-embedded mouse intestine tissue; fixative not specified (datasheet PB9739) |
| Fixation | Image fixative and duration unreported (datasheet PB9739); 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 PB9739); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9739) |
| Primary antibody | Rabbit anti-PRKAB2, 0.5-1μg/ml (datasheet PB9739) |
| Primary incubation | Overnight at 4 °C (datasheet PB9739) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9739) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PRKAB2-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: PB9739). The published protocol also reports EDTA at pH 9.0 or citrate at pH 6.0 (PMC13232994).
Section 2
What Is the Expected PRKAB2 Staining Pattern?
In paraffin IHC, PRKAB2 has a general cytoplasmic pattern, with high staining reported in respiratory, glandular, exocrine glandular, and liver cells (HPA tissue IHC: Enhanced; medium consistency with RNA). A continuous cell-surface rim is unexpected because PRKAB2 has no annotated transmembrane segment (UniProt O43741 topology). ICC-IF has a different reported emphasis: mainly nucleoplasmic, with additional cytosolic localization (HPA subcellular ICC-IF: approved).
| Cytoplasmic staining in hepatocytes, duodenal glandular cells, or bronchial respiratory epithelial cells. | This matches the general cytoplasmic IHC profile and cell types scored High (HPA tissue IHC). Judge signal within identifiable cells; HPA's Enhanced rating carries a medium-consistency caveat relative to RNA data (HPA tissue IHC). |
| A crisp cell-surface rim dominates, with little intracellular staining. | Treat this as a possible staining artefact: the rim conflicts with the general cytoplasmic IHC profile (HPA tissue IHC) and the absence of a transmembrane segment (UniProt O43741 topology). Recheck the control section before assigning PRKAB2 localization. |
| Strong staining appears chiefly in an unexpected cell population rather than the documented high-staining cells. | Consider antibody cross-reactivity or endogenous chromogenic detection activity (general IHC practice). HPA reports high staining in specified cell populations, not every cell in those tissues (HPA tissue IHC); morphology and detection controls matter. |
| Color spreads across stroma, empty areas, or many cell types without clear intracellular boundaries. | Diffuse background cannot establish PRKAB2 expression (general IHC practice). Compare the background with the specific cytoplasmic pattern in documented high-staining cells (HPA tissue IHC), then assess blocking, washing, and detection controls. |
| No cellular signal appears in a section containing well-preserved hepatocytes or pancreatic exocrine glandular cells. | A failed stain is plausible because both cell populations were scored High (HPA tissue IHC). Check tissue quality and the IHC workflow before calling the specimen PRKAB2-negative (general IHC practice); HPA lists no validated negative tissue here (HPA tissue IHC). |
💡Expected PRKAB2 appearanceCall a positive paraffin IHC result when cytoplasmic signal is clear in documented High cells, such as hepatocytes or bronchial respiratory epithelial cells (HPA tissue IHC); isolated surface rims or diffuse extracellular color warrant artefact checks (HPA tissue IHC; UniProt O43741 topology; general IHC practice).
| Evidence strength and tissue choice | The tissue IHC profile is Enhanced, with medium consistency between staining and RNA (HPA tissue IHC). Use an HPA High cell population as a practical positive comparator; skeletal muscle and tongue are RNA-enhanced tissues, not documented High IHC examples in this payload (HPA tissue IHC). |
| IHC and ICC-IF compartment calls | Paraffin tissue IHC is described as generally cytoplasmic (HPA tissue IHC). ICC-IF places PRKAB2 mainly in nucleoplasm and additionally in cytosol (HPA subcellular ICC-IF: approved). Interpret each application against its own observation; this guide gives no ICC-IF protocol. |
| Topology and processing | PRKAB2 has no annotated transmembrane segment or signal peptide, and its annotated chain spans residues 1–272 (UniProt O43741 topology and processing). These annotations support scrutiny of a membrane-rim pattern; they do not identify an antibody epitope or establish how processing affects IHC. |
| Isoforms and modification sites | UniProt lists two isoforms and modified residues, including phosphorylation sites at 39, 40, 69, 95, and 108 (UniProt O43741 isoforms and modified residues). Epitope coverage is not supplied, so neither isoform-specific staining nor modification-dependent signal can be predicted from this record. |
| Situation | Likely cause | Next action |
|---|---|---|
| The documented positive cells show no chromogenic signal. | The staining run or section may have failed; high PRKAB2 staining is reported in hepatocytes and pancreatic exocrine glandular cells (HPA tissue IHC; general IHC practice). | Confirm recognizable positive cells are present, then review retrieval, primary antibody use, detection reagents, and counterstain visibility (general IHC practice). Do not infer target-specific retrieval sensitivity from HPA. |
| Background obscures cell boundaries throughout the section. | Incomplete blocking, washing, or detection control can produce diffuse color (general IHC practice). HPA's expected tissue pattern is general cytoplasmic expression (HPA tissue IHC). | Inspect a detection-only control and review blocking, washes, and reagent handling (general IHC practice). Score only intracellular signal that remains distinguishable from background. |
| A strong rim outlines cells, but their cytoplasm is faint. | A membrane-dominant result conflicts with the reported cytoplasmic IHC pattern and lack of a transmembrane segment (HPA tissue IHC; UniProt O43741 topology). | Compare the rim with a documented High-cell control and the detection-only control; review morphology and staining deposits before interpreting it as PRKAB2 (HPA tissue IHC; general IHC practice). |
| Signal is concentrated in cells other than the documented High populations. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA High calls specify particular cells within each tissue (HPA tissue IHC). | Verify cell identity by morphology and compare detection controls and an HPA High-cell section (HPA tissue IHC; general IHC practice). Avoid declaring an unlisted population positive from color alone. |
| The paraffin section is nuclear-only, while an ICC-IF image also looks nuclear. | ICC-IF reports mainly nucleoplasmic PRKAB2, but tissue IHC reports a general cytoplasmic pattern (HPA subcellular ICC-IF: approved; HPA tissue IHC). The assays have distinct observations. | Evaluate the paraffin result against tissue IHC controls and morphology; review retrieval and detection workflow if cytoplasmic signal is absent (HPA tissue IHC; general IHC practice). Do not use ICC-IF localization as an IHC acceptance rule. |
| Low staining is mistaken for a validated negative control. | HPA lists low staining in several cell populations but supplies no negative tissue in this record (HPA tissue IHC). Low expression is an uncertain absence-of-signal benchmark. | Label low-staining cells as low comparators only; include a documented High population and an appropriate detection control when judging assay performance (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PRKAB2 staining
PB9739 · IHCIHC analysis of AMPK beta 2 using anti-AMPK beta 2 antibody (PB9739). AMPK beta 2 was detected in a paraffin-embedded section of mouse intes
PB9739 · IHCIHC analysis of AMPK beta 2 using anti-AMPK beta 2 antibody (PB9739). AMPK beta 2 was detected in a paraffin-embedded section of rat intesti
PB9739 · IFIF analysis of AMPK beta 2 using anti-AMPK beta 2 antibody (PB9739). AMPK beta 2 was detected in immunocytochemical section of A431 cells. E
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000131791-PRKAB2/tissue/cerebral+cortex#imid_12830739
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000131791-PRKAB2/tissue/colon#imid_12830657
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000131791-PRKAB2/tissue/liver#imid_12830658
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000131791-PRKAB2/tissue/kidney#imid_12830659
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044342; green = PRKAB2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044342; green = PRKAB2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PRKAB2 IHC & IF
🧪Run bronchus first and score respiratory epithelial cells, which show High PRKAB2 staining (HPA: High in bronchus respiratory epithelial cells). HPA detects PRKAB2 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and expect any cells judged negative within the positive section to show counterstain without specific chromogen, while recognizing that the supplied HPA rows establish no antigen-negative cell population (HPA: no negative rows).
Positive control tissue: Bronchus (Respiratory epithelial cells, HPA High)
Negative control tissue: None in HPA: PRKAB2 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 PRKAB2 in A-431, SK-MEL-30, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), KOLF2.1J, NIH 3T3, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a secondary-only slide and a host- and class-matched control IgG in place of the rabbit primary antibody (IHC caption: rabbit anti-PRKAB2); use PRKAB2-knockout material, if available, as a biological negative. For chromogenic detection, block endogenous peroxidase and check endogenous biotin background when using the caption’s streptavidin–biotin detection method (IHC caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9739 paraffin-section caption does not state the fixative (IHC caption: fixative unreported). The caption uses heat retrieval in EDTA pH 8.0, but supplies no comparison establishing that retrieval is required (IHC caption: heat-mediated EDTA retrieval). No supplied evidence establishes whether frozen sections or IF are easier than paraffin IHC; for bronchus, assess mucus-associated background during scoring (standard IHC practice).
HPA tissue IHC evidence for PRKAB2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PRKAB2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PRKAB2 IHC Tips
Troubleshoot PRKAB2 staining in paraffin sections by checking retrieval, cellular distribution, detection background and scoring against tissue controls.
Which retrieval conditions should I try first for PRKAB2 in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA pH 8.0 for paraffin-section PRKAB2 IHC (datasheet PB9739). The documented mouse intestine image used this retrieval before incubation with 1 μg/ml primary antibody overnight at 4°C (datasheet PB9739). If staining is weak, first check heating consistency and section adhesion, then compare a shorter or longer retrieval time on matched sections while holding detection conditions constant (standard IHC practice). Excessive heating can damage morphology or increase background, so judge each condition by both cellular staining and tissue preservation (standard IHC practice). Do not attribute a retrieval difference to fixation without a recorded fixation method (datasheet PB9739: fixative unstated).
How should I troubleshoot weak staining when the section fixative is unknown?
Target-specific fixation sensitivity is unknown: the PB9739 image describes a paraffin section but does not state its fixative (datasheet PB9739). Record the actual fixative, fixation duration and processing history for each section before comparing staining intensity (standard IHC practice). Run matched sections through the same EDTA pH 8.0 retrieval and antibody incubation, changing one preparation variable at a time (datasheet PB9739; standard IHC practice). If a well-preserved control stains while the test section does not, assess tissue handling and antigen preservation alongside antibody performance (standard IHC practice). Neither the tissue staining profile nor PRKAB2 sequence annotations establish a preferred fixative (HPA: tissue IHC; UniProt O43741).
Should nuclear PRKAB2 staining be rejected in chromogenic IHC?
Do not reject nuclear staining solely because the tissue IHC profile describes general cytoplasmic expression: cell imaging places PRKAB2 mainly in the nucleoplasm and additionally in the cytosol (HPA: tissue IHC; HPA: subcellular). UniProt supplies no subcellular annotation and reports no transmembrane segment, so it cannot settle a compartment discrepancy in a section (UniProt O43741). Compare nuclear and cytoplasmic signals within identifiable, intact cells and against the same compartments in a control section (standard IHC practice). Check whether nuclear color follows hematoxylin intensity, tissue edges or precipitate rather than cell boundaries before assigning it to PRKAB2 (standard IHC practice).
Could isoforms or epitope masking explain inconsistent PRKAB2 staining?
PRKAB2 has 2 annotated isoforms, but the supplied antibody caption does not map the recognized epitope or establish isoform selectivity (UniProt O43741; datasheet PB9739). The protein also has reported modified residues, including phosphorylation at positions 39, 40, 69, 95 and 108 (UniProt O43741). Those annotations alone do not show that a modification masks this antibody's epitope after tissue processing (UniProt O43741; datasheet PB9739). Compare matched sections with consistent retrieval and detection before proposing a biological difference, and seek epitope mapping or independent validation if isoform specificity matters (standard IHC practice). Report the result as PRKAB2 immunoreactivity until that specificity is established (standard IHC practice).
How can IF help resolve an ambiguous chromogenic PRKAB2 pattern?
Use a separate IF/ICC validation experiment to compare PRKAB2 with a marker for the expected cell type, such as an epithelial marker when examining glandular cells (HPA: high in duodenal glandular cells; standard IF practice). HPA cell imaging places PRKAB2 mainly in the nucleoplasm, with additional cytosolic signal, which provides compartments to inspect alongside the chromogenic section (HPA: subcellular). Choose a fluorophore channel with low measured tissue autofluorescence and include single-stain controls before interpreting overlap (standard IF practice). Because PRKAB2 lacks a transmembrane segment, test permeabilisation for access to intracellular staining, while treating the antibody epitope's exact location as unknown (UniProt O43741; datasheet PB9739; standard IF practice).
What should I check when DAB background obscures PRKAB2 staining?
The documented paraffin-section method uses a biotinylated secondary antibody, a streptavidin–biotin complex and DAB, so inspect each detection stage when color appears outside cells (datasheet PB9739). Include a no-primary control and check endogenous peroxidase blocking before assigning diffuse brown signal to PRKAB2 (standard IHC practice). If background remains, assess endogenous biotin with a suitable detection control, particularly because this method uses biotin-based amplification (datasheet PB9739; standard IHC practice). The example used 10% goat serum blocking and 1 μg/ml primary antibody; adjust blocking, washes or primary concentration on matched sections while monitoring positive tissue (datasheet PB9739; standard IHC practice).
How should PRKAB2 IHC be scored across sections with different cellular composition? ⚠ ANSWER MARKED FOR VERIFICATION
Score identified cell populations separately, since reported high staining spans respiratory epithelial cells, glandular cells, hepatocytes and pancreatic exocrine glandular cells (HPA: tissue IHC). For intensity based reporting, use a predefined H-score or record the percentage of positive cells within each annotated population; retain nuclear and cytoplasmic scores separately when both are visible (standard IHC practice; HPA: subcellular). Normalize counts to the number of evaluable cells, or report positive-cell density per mm² of viable tissue when area is the intended denominator (standard IHC practice). Keep retrieval, imaging, thresholds and background subtraction consistent across sections, and exclude damaged or necrotic regions before comparison (standard IHC practice).
How can I distinguish true PRKAB2 signal from section artefact?
A plausible result has reproducible cellular staining in intact structures and agrees with an appropriate control, rather than appearing only at section edges or in necrotic areas (standard IHC practice). High staining has been reported in duodenal glandular cells and hepatocytes, while caudate neuronal cells are listed as low, so interpret cell identity before comparing intensity (HPA: tissue IHC). Cytoplasmic tissue staining is reported, and nucleoplasmic plus cytosolic localization appears in cell imaging; a discrepant compartment warrants validation rather than automatic rejection (HPA: tissue IHC; HPA: subcellular). Check no-primary and detection controls for endogenous peroxidase or biotin signal before calling DAB color PRKAB2 positive (datasheet PB9739; standard IHC practice).
Boster reagents
Best PRKAB2 / 5'-AMP-activated protein kinase subunit beta-2 IHC Antibodies
PB9739 has real paraffin-section IHC images from mouse and rat intestine and human lung cancer tissue, plus an IF/ICC image from A431 cells (catalog image captions).
Real IHC data
Anti-AMPK beta 2/PRKAB2 Antibody ®
Cat # PB9739
PB9739 is listed for human, mouse and rat IHC and for human IF/ICC; its captions show paraffin-section IHC in mouse and rat intestine and human lung cancer tissue, and IF/ICC in A431 cells (catalog applications/reactivity; image captions). M05077 is listed for human frozen-section IHC and ICC, with no supplied IHC or IF figure (catalog reactivity/dilution_raw; image captions).
Which to pick: Choose PB9739 for paraffin-section tissue IHC: its own captions document EDTA pH 8 retrieval and 1 μg/ml primary antibody; the fixative is unreported (PB9739 IHC image captions). Choose PB9739 for IF/ICC based on its A431 image at 2 μg/ml; M05077 is a human-reactive mouse monoclonal clone 6G1 listed for ICC and frozen-section IHC without a supplied figure (PB9739 IF image caption; M05077 catalog clone/reactivity/dilution_raw/image captions). For cross-species IHC, PB9739 is listed for human, mouse and rat, while M05077 is listed for human only (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43741 (AAKB2_HUMAN, 5'-AMP-activated protein kinase subunit beta-2).
- Human Protein Atlas. PRKAB2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PRKAB2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. PRKAB2 antibody validation summary (1 antibodies).
- PRKAB2 as a tumor suppressor in renal cell carcinoma: inhibiting mitophagy via the LRPPRC-PRKN/parkin interaction and cardiolipin biosynthesis. Autophagy 2026 — PMC13232994.
- PubMed PMID:9575201 — UniProt-cited evidence.
- PubMed PMID:12490143 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PRKAB2 (5'-AMP-activated protein kinase subunit beta-2). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9739). Validated IHC antibody: PB9739 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Respiratory epithelial cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse intestine tissue; fixative not specified (datasheet PB9739); 2. Fixation: Image fixative and duration unreported (datasheet PB9739); 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 PB9739); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9739); 8. Primary antibody: Rabbit anti-PRKAB2, 0.5-1μg/ml (datasheet PB9739); 9. Primary incubation: Overnight at 4 °C (datasheet PB9739). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show only medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; confirm epitope coverage (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt O43741, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet PB9739, PMC13232994. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PRKAB2/immunohistochemistry-ihc-guide.