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PFKP / ATP-dependent 6-phosphofructokinase, platelet type · IHC design guide
Design Immunohistochemistry for PFKP
Plan PFKP chromogenic IHC on paraffin sections using the catalog antibody’s documented conditions. Expect cytoplasmic staining (HPA tissue IHC), and interpret intensity cautiously because tissue staining has low concordance with RNA data (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PFKP 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in glandular and endothelial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07337-3) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Tissue staining has low concordance with RNA data (HPA tissue IHC) | |
| Regulation | Staining regulation is not established (UniProt) | |
| Isoform / epitope | 2 isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended PFKP IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet A07337-3). Three published PFKP IHC protocols provide comparisons (PMC13502873; PMC7259711; PMC7210009).
| Sample | Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A07337-3) |
| Fixation | Image fixative and duration unreported (datasheet A07337-3); 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 A07337-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07337-3) |
| Primary antibody | Rabbit anti-PFKP, 2-5 μg/ml (datasheet A07337-3) |
| Primary incubation | Overnight at 4 °C (datasheet A07337-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07337-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PFKP-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A07337-3); citrate pH 6.0 was used with a different antibody (PMC7210009 methods).
Section 2
What Is the Expected PFKP Staining Pattern?
PFKP is a cytoplasmic enzyme without a transmembrane segment, so interpretable staining should favor cell cytoplasm (UniProt Q01813: subcellular location and topology). HPA reports high IHC staining in several glandular, respiratory epithelial, glial, and endothelial cell populations, with a general cytoplasmic profile (HPA: tissue IHC). Treat these as provisional expectations: HPA rates the tissue staining Uncertain because antibody staining and RNA expression show low consistency (HPA: tissue IHC reliability).
| Cytoplasmic staining in adrenal or breast glandular cells, or bronchial respiratory epithelium (HPA: High in each population). | This fits the reported tissue pattern and PFKP location (HPA: tissue IHC; UniProt Q01813: Cytoplasm). Judge the relevant cells against nearby tissue and controls; intensity alone cannot establish specificity because HPA rates its tissue IHC evidence Uncertain (HPA: reliability). |
| A dominant nuclear, membrane-rim, or extracellular deposit replaces the cytoplasmic pattern. | That distribution conflicts with the reported cytoplasmic location and absence of a transmembrane segment (UniProt Q01813: location and topology). Check morphology and controls before calling it PFKP; misplaced chromogen can reflect nonspecific staining or a detection artifact (general IHC practice). |
| Strong staining appears chiefly in an unexpected cell population, while the reported population is faint. | Compare cell identity and distribution with the HPA tissue entry before scoring (HPA: tissue IHC). Cross-reactivity or endogenous detection activity is possible, but HPA's Uncertain rating means an unfamiliar pattern cannot be dismissed solely because it differs from its reference image (HPA: reliability; general IHC practice). |
| Diffuse color obscures tissue boundaries or coats cells with no clear cytoplasmic contrast. | This is difficult to assign to PFKP's reported cytoplasmic compartment (UniProt Q01813: Cytoplasm). Review no-primary controls, detection chemistry, and wash and blocking conditions; widespread deposition can arise from nonspecific reagent binding or endogenous enzyme activity (general IHC practice). |
| No visible staining occurs in a section containing a reported high-staining population. | Adrenal glandular cells and bronchial respiratory epithelial cells are reported High, but HPA rates the underlying tissue pattern Uncertain (HPA: tissue IHC). First establish that the run and tissue are interpretable; a blank slide alone cannot distinguish low assay sensitivity from a reference-pattern discrepancy (general IHC practice; HPA: reliability). |
💡Expected PFKP appearanceCall a result consistent when cytoplasmic color is clear in a reported High cell population, such as adrenal glandular cells (UniProt Q01813: Cytoplasm; HPA: High), while dominant nuclear or membrane-rim color warrants artifact review (UniProt Q01813: location and topology; general IHC practice).
| Cell and tissue context | HPA reports High staining in multiple populations, including glandular, respiratory epithelial, glial, and endothelial cells; hepatocytes are Low and adipocytes Not detected (HPA: tissue IHC). Score the named cells within each tissue rather than treating a whole section as uniformly positive or negative (general IHC practice). |
| Evidence strength and antibody choice | The tissue IHC profile is Uncertain, and HPA018257 has an Uncertain IHC status (HPA: reliability; HPA: antibodies). A matching distribution is supportive, not definitive; compare controls and independent evidence when specificity matters (general IHC practice). |
| Protein compartment and processing | UniProt places PFKP in cytoplasm, lists no transmembrane segment or signal peptide, and identifies a chain spanning residues 1–784 (UniProt Q01813: location, topology, processing). These annotations support a cytoplasmic reading; they do not identify an antibody epitope or predict retrieval performance. |
| Isoforms and epitope limits | UniProt lists two PFKP isoforms, but the supplied record does not map the IHC antibody epitope (UniProt Q01813: isoforms; HPA: antibodies). Therefore, a difference between specimens cannot be assigned to isoform recognition from these sources alone. |
| IF/ICC Q&A: where should signal appear? | Expect cytosolic signal: HPA reports a supported cytosol location in ICC-IF and marks HPA056484 ICC Approved (HPA: subcellular; HPA: antibodies). That ICC assessment does not raise the separate tissue IHC profile above Uncertain (HPA: tissue IHC reliability). |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported High cell population shows no chromogenic signal. | The run may lack usable sensitivity, or the chosen specimen may not reproduce HPA's Uncertain tissue pattern (general IHC practice; HPA: reliability). | Check section integrity and the run's controls, then review retrieval and detection settings as general IHC variables; no PFKP-specific retrieval condition is supplied (general IHC practice). |
| Color is concentrated in nuclei or along cell borders. | The distribution disagrees with PFKP's cytoplasmic location and lack of a transmembrane segment (UniProt Q01813: location and topology). | Inspect the corresponding tissue morphology and no-primary control; adjust detection or blocking only if controls point to an assay artifact (general IHC practice). |
| Diffuse background hides the expected cytoplasm. | Nonspecific reagent binding, residual detection activity, or excessive chromogen development can reduce contrast (general IHC practice). | Compare a no-primary control, confirm the detection system's blocking steps, and optimize washes and development time (general IHC practice). |
| Unexpected cells stain more strongly than the reported cell population. | Cross-reactivity or endogenous detection activity may contribute, but HPA's tissue pattern is Uncertain (general IHC practice; HPA: reliability). | Identify the stained cells on the section, compare the relevant HPA tissue entry, and use appropriate negative controls before assigning PFKP positivity (HPA: tissue IHC; general IHC practice). |
| Hepatocytes stain weakly or adipocytes show no signal. | Those findings match the supplied Low and Not detected observations, respectively (HPA: tissue IHC). | Do not use either population alone to declare assay failure; assess a reported High population in a suitable control section while retaining HPA's Uncertain reliability caveat (HPA: tissue IHC; general IHC practice). |
| An IF/ICC image and a paraffin IHC section appear different. | HPA supports cytosolic ICC-IF localization but rates tissue IHC staining Uncertain; the antibody assessments also differ by application (HPA: subcellular; HPA: tissue IHC; HPA: antibodies). | Interpret each image with its own application controls and cell context; use the shared cytoplasmic location as the compartment check (UniProt Q01813: Cytoplasm; general IHC/IF practice). |
Experiment evidence
IHC & IF images showing PFKP staining
A07337-3 · IHCIHC analysis of PFKP using anti-PFKP antibody (A07337-3). PFKP was detected in a paraffin-embedded section of human colon adenocarcinoma tis
A07337-3 · IHCIHC analysis of PFKP using anti-PFKP antibody (A07337-3). PFKP was detected in a paraffin-embedded section of human diffuse large B cell lym
A07337-2 · IHCIHC analysis of PFKP using anti-PFKP antibody (A07337-2). PFKP was detected in a paraffin-embedded section of human breast cancer tissue. He
A07337-2 · IHCIHC analysis of PFKP using anti-PFKP antibody (A07337-2). PFKP was detected in a paraffin-embedded section of human uterine tumor tissue. He
A30471 · IHCImmunohistochemical analysis of paraffin-embedded Human testis. Antibody was diluted at 1:100(4° overnight). High-pressure and temperature T
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000067057-PFKP/tissue/cerebral+cortex#imid_5000388
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000067057-PFKP/tissue/colon#imid_4988854
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000067057-PFKP/tissue/liver#imid_4988855
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000067057-PFKP/tissue/kidney#imid_4988864
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA056484; green = PFKP, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA056484; green = PFKP, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PFKP IHC & IF
🧪Run bronchus first: respiratory epithelial cells should stain strongly (HPA: High in bronchus respiratory epithelial cells). Run adipose tissue as the negative tissue (HPA: Not detected in adipocytes); adjacent non-epithelial cells on the bronchus slide can indicate background if unstained, but the supplied HPA row does not establish them as PFKP-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PFKP in HeLa, MCF-7, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a species- and clonality-matched isotype control, and PFKP-knockout tissue or cells as a biological negative (standard IHC practice). Quench endogenous peroxidase and inspect bronchial mucus for nonspecific DAB deposits before scoring epithelial staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A07337-3 caption does not state a fixative (selected-SKU tissue-IHC caption). That caption used heat retrieval in EDTA at pH 8.0, so retrieval is documented for that paraffin-section example, but its necessity has not been established across specimens (selected-SKU tissue-IHC caption). IF/ICC can assess the reported cytosolic location with permeabilization (HPA: supported cytosol; standard IF practice); the supplied evidence does not establish whether IF or frozen sections are easier than paraffin IHC, and bronchial mucus warrants a nonspecific-staining check (standard IHC practice).
HPA tissue IHC evidence for PFKP
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PFKP IHC Tips
PFKP is a cytoplasmic glycolytic enzyme; interpret paraffin-section staining with attention to retrieval, cell identity and assay controls (UniProt Q01813; HPA tissue IHC).
What retrieval should I try first when PFKP staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A07337-3). The catalog antibody produced a PFKP image in human colon adenocarcinoma after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A07337-3). If signal remains weak, compare retrieval durations on adjacent sections while holding antibody concentration and detection conditions constant (standard IHC practice). Include a no-primary control and inspect tissue preservation, since stronger heating can increase background or damage morphology without establishing that a new signal is PFKP (standard IHC practice).
How should I troubleshoot possible fixation effects on PFKP staining?
The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity for this antibody is unknown (datasheet A07337-3). Record each specimen’s fixative, fixation duration and processing history before comparing staining across blocks (standard IHC practice). When signal differs between blocks, stain sections together using the reported EDTA pH 8.0 retrieval and 2 μg/ml primary concentration to reduce workflow variation (datasheet A07337-3; standard IHC practice). Review morphology and a no-primary control alongside PFKP staining; a change in intensity alone cannot establish that fixation altered its epitope (standard IHC practice).
Where should convincing PFKP staining appear in a paraffin section?
Expect predominantly cytoplasmic staining because PFKP is annotated in the cytoplasm and has no transmembrane segment (UniProt Q01813). The tissue IHC profile also describes general cytoplasmic expression, although its reliability is uncertain because staining and RNA data have low consistency (HPA tissue IHC). Assess signal within identifiable cells rather than treating diffuse deposits outside cell boundaries as positive staining (standard IHC practice). If staining appears exclusively nuclear or outlines membranes, review the no-primary control, counterstain and section morphology before assigning it to PFKP (UniProt Q01813; standard IHC practice).
Could isoforms or epitope masking explain different PFKP staining patterns?
PFKP has 2 annotated isoforms, but the supplied antibody evidence does not identify its epitope or establish isoform recognition (UniProt Q01813; datasheet A07337-3). Several modified residues are annotated, including phosphoserines at positions 6, 12, 21 and 142; their effect on this antibody’s staining is unknown (UniProt Q01813). Compare adjacent sections under the reported EDTA pH 8.0 retrieval before attributing variable signal to an isoform or modification (datasheet A07337-3; standard IHC practice). For an isoform-specific conclusion, obtain epitope-mapping or isoform-specific validation evidence rather than relying on chromogenic intensity alone (standard IHC practice).
How can I check PFKP localisation by multiplex immunofluorescence?
Use IF/ICC as a separate assay: HPA reports supported cytosolic localisation and lists ICC/IF images, while the selected antibody evidence describes paraffin-section IHC (HPA subcellular; datasheet A07337-3). In colon, a multiplex panel could pair PFKP with an endothelial marker such as CD31 to examine the cell population reported as high in tissue IHC (HPA tissue IHC; standard IF practice). Choose a far-red fluorophore when tissue autofluorescence obscures shorter wavelengths, and include single-label controls (standard IF practice). Because PFKP is cytosolic with no transmembrane segment, permeabilise cells or sections to allow antibody access to intracellular epitopes, then optimise permeabilisation for the specimen (UniProt Q01813; standard IF practice).
How do I separate PFKP signal from chromogenic background?
The selected example used goat-serum blocking, an anti-rabbit secondary and DAB development after overnight primary incubation (datasheet A07337-3). Include no-primary and secondary-only controls to reveal detection-reagent background, and apply a peroxidase block before HRP detection as a general chromogenic IHC step (standard IHC practice). Compare suspected staining with section edges, folds and damaged areas, where reagent trapping can create misleading deposits (standard IHC practice). If background persists, adjust blocking, washes or primary concentration in controlled comparisons, using the reported 2 μg/ml as the starting reference rather than interpreting darker DAB as greater specificity (datasheet A07337-3; standard IHC practice).
What is a defensible way to quantify PFKP IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue compartment before scoring, since HPA reports high staining in colon endothelial cells and an uncertain overall tissue-IHC profile (HPA tissue IHC). For identifiable cells, record the percentage at each intensity grade 0–3 and calculate an H-score from 0–300, or report the percentage of positive cells using a prespecified threshold (standard IHC practice). For dispersed positive cells, density per mm² of viable tissue may be more useful (standard IHC practice). Normalise comparisons to the same scored cell type and viable tissue area, and keep retrieval, imaging and scoring rules consistent across sections (standard IHC practice).
When should an apparent PFKP-positive region be treated cautiously?
A credible signal should occupy cell cytoplasm, consistent with PFKP’s annotated location and HPA’s general tissue staining pattern (UniProt Q01813; HPA tissue IHC). Check cell identity: HPA reports high staining in colon endothelial cells, while the selected catalog image shows staining in a human colon adenocarcinoma section without establishing every positive cell type (HPA tissue IHC; datasheet A07337-3). Discount staining concentrated at section edges, folds or necrotic regions until morphology and controls support it (standard IHC practice). Residual endogenous peroxidase can produce DAB signal, so compare peroxidase-blocked and no-primary controls before calling an unexpected compartment or cell population PFKP-positive (standard IHC practice).
Boster reagents
Best PFKP / ATP-dependent 6-phosphofructokinase, platelet type IHC Antibodies
Three PFKP antibodies have human paraffin-section IHC images (catalog image captions). A07337-2 also has mouse and rat IHC images; A07337-2 and A30471 list IF/ICC applications (catalog image captions; catalog applications).
Real IHC data
Anti-PFKP Antibody ®
Cat # A07337-3
Real IHC data
Anti-PFKP Antibody
Cat # A07337-2
Real IHC data
Anti-K6PP Antibody
Cat # A30471
A07337-3 shows IHC in human colon adenocarcinoma; A07337-2 shows IHC in human breast cancer and mouse and rat brain (respective image captions). A30471 shows IHC in human testis and lists IF/ICC among its applications (A30471 image caption; catalog applications).
Which to pick: For human paraffin-section chromogenic IHC, choose A07337-3: its image caption documents EDTA pH 8 retrieval, 2 μg/ml primary antibody and DAB detection (A07337-3 image caption). For IF/ICC or human, mouse and rat IHC, choose polyclonal A07337-2, which lists IF/ICC and has IHC images for all three species (A07337-2 dilution_raw; catalog applications; image captions). A30471 is the polyclonal option that lists monkey reactivity, though its IHC image documents only human paraffin-section testis; fixation is unreported in all three SKUs’ image captions (A30471 dilution_raw; catalog reactivity; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q01813 (PFKAP_HUMAN, ATP-dependent 6-phosphofructokinase, platelet type).
- Human Protein Atlas. PFKP tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. PFKP subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. PFKP antibody validation summary (2 antibodies).
- Platelet-Type Phosphofructokinase Drives Immune Checkpoint Coexpression and Metabolic Pathway Activation in Liver Cancer: The Cancer Genome Atlas-Based and Experimental Validation. Clinical and translational gastroenterology 2026 — PMC13502873.
- PFKP is transcriptionally repressed by BRCA1/ZBRK1 and predicts prognosis in breast cancer. PloS one 2020 — PMC7259711.
- Mechanical regulation of glycolysis via cytoskeleton architecture. Nature 2020 — PMC7210009.
- PFKP Activation Ameliorates Foot Process Fusion in Podocytes in Diabetic Kidney Disease. Frontiers in endocrinology 2021 — PMC8794994.
- PubMed PMID:8117307 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PFKP (ATP-dependent 6-phosphofructokinase, platelet type). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07337-3). Validated IHC antibody: A07337-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A07337-3); 2. Fixation: Image fixative and duration unreported (datasheet A07337-3); 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 A07337-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07337-3); 8. Primary antibody: Rabbit anti-PFKP, 2-5 μg/ml (datasheet A07337-3); 9. Primary incubation: Overnight at 4 °C (datasheet A07337-3). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Tissue staining has low concordance with RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; check antibody epitope coverage (UniProt); Regulation: Staining regulation is not established (UniProt). Evidence: UniProt Q01813, HPA tissue IHC · Uncertain, HPA ICC-IF · cytosol., Datasheet A07337-3, PMC13502873, PMC7259711. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PFKP/immunohistochemistry-ihc-guide.