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IPMK / Inositol polyphosphate multikinase · IHC design guide
Design Immunohistochemistry for IPMK
Plan paraffin-section IPMK staining using the catalog antibody’s IHC dilution range of 1:100–1:300 (datasheet A06955-1). Assess cytoplasmic tissue staining while keeping the nuclear molecular annotation and the uncertain tissue IHC reliability in view (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IPMK 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 in tissue (HPA tissue IHC); nuclear molecularly (UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A06955-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
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 awaits external verification (HPA tissue IHC) | |
| Regulation | Highest expression in skeletal muscle and liver (UniProt) | |
| Isoform / epitope | No annotated isoforms; one chain spans residues 2–416 (UniProt) |
Section 1
Recommended IPMK IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A06955-1) with a published IPMK chromogenic IHC protocol (PMC10197106).
| Sample | Paraffin-embedded human Colon cancer tissue; fixative not specified (datasheet A06955-1) |
| Fixation | Image fixative and duration unreported (datasheet A06955-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A06955-1); 20 min, 95–100 °C (standard) |
| 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-IPMK, 1:100-1:300 (datasheet A06955-1) |
| 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 | IPMK-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 9.0 for the catalog antibody (datasheet: A06955-1); the published protocol does not specify retrieval (PMC10197106).
Section 2
What Is the Expected IPMK Staining Pattern?
IPMK is a nuclear protein without a transmembrane segment (UniProt Q8NFU5). HPA ICC-IF supports nucleoplasmic localisation, while HPA tissue IHC reports cytoplasmic staining in most tissues (HPA: subcellular; HPA: tissue IHC). Expect staining in identifiable cells, including glandular, respiratory epithelial and neuronal cells reported at medium intensity (HPA: tissue IHC). Interpret the IHC pattern cautiously: its tissue-level reliability is Uncertain, pending external verification (HPA: tissue IHC).
| Nuclear staining in identifiable cells, with possible cytoplasmic staining on the IHC section. | Nuclear localisation agrees with UniProt and supported nucleoplasmic ICC-IF staining (UniProt Q8NFU5; HPA: subcellular). Cytoplasmic tissue staining has been reported, but its IHC reliability is Uncertain; record nuclear and cytoplasmic scores separately rather than treating either compartment as definitive (HPA: tissue IHC). |
| Medium staining in bronchial respiratory epithelium, cerebellar Purkinje cells or cortical neurons. | These are reported positive cell populations at Medium intensity (HPA: tissue IHC). Check that staining follows the named cells rather than every structure on the section (general IHC practice). A matching distribution supports interpretation, but it cannot independently resolve the Uncertain IHC reliability (HPA: tissue IHC). |
| A sharp membrane rim or extracellular deposit dominates the apparent signal. | That distribution conflicts with the nuclear annotation and absence of a transmembrane segment (UniProt Q8NFU5). Suspect nonspecific staining or a detection artefact; compare cell morphology and controls before assigning an IPMK score (general IHC practice). Cytoplasmic staining alone is less decisive because HPA reports it in tissue IHC (HPA: tissue IHC). |
| Strong staining appears in adipocytes or placental trophoblastic cells. | Both populations are reported Not detected by HPA tissue IHC, so strong staining there warrants a specificity check (HPA: tissue IHC). Cross-reactivity or endogenous chromogen-generating activity are possible causes (general IHC practice). A Not detected HPA result is not proof that every specimen must remain negative, particularly given Uncertain IHC reliability (HPA: tissue IHC). |
| A diffuse brown haze obscures cell boundaries, or a reported positive population has no visible signal. | Haze that does not follow cells is background, not a scorable IPMK pattern (general IHC practice). An absent signal in a reported Medium population may reflect assay performance or sample variation; the HPA observation alone does not diagnose the cause (HPA: tissue IHC). Compare a same-run control before interpreting a negative result (general IHC practice). |
💡Expected IPMK appearanceCall a section positive when identifiable cells show reproducible nuclear staining consistent with UniProt and HPA ICC-IF, while recording any cytoplasmic IHC staining separately under HPA's Uncertain reliability (UniProt Q8NFU5; HPA: subcellular; HPA: tissue IHC). Medium staining is reported in selected cell populations; a dominant membrane rim or cell-independent brown deposit is suspicious (HPA: tissue IHC; general IHC practice).
| Compartment evidence | UniProt places IPMK in the nucleus, and HPA ICC-IF supports nucleoplasmic localisation; HPA tissue IHC instead describes cytoplasmic expression in most tissues with Uncertain reliability (UniProt Q8NFU5; HPA: subcellular; HPA: tissue IHC). Preserve this discrepancy when scoring. |
| Cell population and specimen choice | HPA reports Medium staining in several glandular, respiratory epithelial and neuronal populations, but Not detected in specific populations such as adipocytes and trophoblastic cells (HPA: tissue IHC). Use the reported cell type within the tissue, not the tissue name alone, to assess agreement. |
| Antibody validation | HPA037837 is Uncertain for IHC and Supported for ICC; HPA057542 has no IHC status listed and is Supported for ICC (HPA: antibodies). Supported ICC staining does not establish that either antibody yields a specific chromogenic IHC pattern (HPA: antibodies). |
| Topology and processing | No transmembrane segment or signal peptide is annotated, and no glycosylation sites are listed (UniProt Q8NFU5). These annotations give no basis to expect a membrane or secreted staining pattern; they do not establish epitope accessibility or fixation sensitivity (UniProt Q8NFU5). |
| IF/ICC Q&A: Where should IPMK appear? | Mainly in the nucleoplasm, with an additional basal-body location reported by HPA ICC-IF; the main location is Supported and the additional location Approved (HPA: subcellular). That cell-image evidence should be kept distinct from the Uncertain tissue IHC profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported Medium-positive cells have no chromogenic signal. | The section or detection run may have failed, or that specimen may differ from the reported examples (general IHC practice; HPA: tissue IHC). | Check a same-run reported positive cell population and the detection controls; review antigen-retrieval and antibody-dilution records before calling the sample negative (general IHC practice). |
| Brown staining covers cells and empty spaces alike. | Cell-independent colour is consistent with background from the detection workflow, rather than a localised IPMK pattern (general IHC practice). | Inspect a no-primary control, wash and blocking conditions, and endogenous peroxidase quenching; score only signal that follows identifiable cells (general IHC practice). |
| A strong membrane rim is scored as IPMK. | The pattern disagrees with nuclear localisation and the lack of a transmembrane segment (UniProt Q8NFU5). | Recheck morphology and no-primary controls, then assess nuclear and cytoplasmic compartments separately; flag persistent rim staining for specificity review (general IHC practice). |
| Adipocytes or trophoblastic cells stain strongly. | Those cells are reported Not detected, though the HPA tissue IHC profile has Uncertain reliability (HPA: tissue IHC). | Check whether colour follows the named cells, inspect no-primary and endogenous-activity controls, and seek independent specificity evidence before treating it as IPMK (general IHC practice). |
| Cytoplasmic staining conflicts with the expected nuclear location. | HPA tissue IHC reports cytoplasmic expression, whereas UniProt and supported HPA ICC-IF place IPMK in the nucleus or nucleoplasm (HPA: tissue IHC; UniProt Q8NFU5; HPA: subcellular). | Report both compartments and the evidence conflict; avoid declaring cytoplasmic IHC signal either confirmed IPMK or an artefact on localisation alone (HPA: tissue IHC; HPA: subcellular). |
| Staining differs markedly between sections in one run. | Uneven section handling or detection can alter the apparent pattern (general IHC practice); HPA supplies no target-specific fixation-sensitivity finding here (HPA: tissue IHC). | Compare same-run controls and review section, retrieval, detection and counterstain records before attributing the difference to IPMK biology (general IHC practice). |
Experiment evidence
IHC & IF images showing IPMK staining
A06955-1 · IHCImmunohistochemical analysis of paraffin-embedded human Colon cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000151151-IPMK/tissue/cerebral+cortex#imid_9233360
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000151151-IPMK/tissue/colon#imid_9233294
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000151151-IPMK/tissue/liver#imid_9233279
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000151151-IPMK/tissue/kidney#imid_9233288
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037837; green = IPMK, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037837; green = IPMK, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IPMK IHC & IF
🧪Run breast first and score its glandular cells for IPMK staining (HPA: Medium in breast glandular cells). Use adipose tissue as the negative comparator, scoring adipocytes (HPA: Not detected in adipocytes); where adipocytes are present on the breast slide, they should remain near background (HPA: Not detected in adipocytes).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show IPMK in A-431, U-251MG, U2OS, Rh30, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a concentration-matched isotype control selected for the primary antibody’s host species and clonality; where available, use IPMK knockout material or an immunizing-peptide block to assess specificity (standard IHC practice). Block endogenous peroxidase and inspect glandular cells and adjacent adipocytes for nonspecific chromogen deposition (standard chromogenic IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected-SKU paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption). That caption used Tris-EDTA at pH 9.0 for retrieval in colon cancer, but it does not establish retrieval dependence in breast tissue (selected-SKU tissue-IHC caption). Frozen sections or IF cannot be judged easier from these data; for IF/ICC, assess nucleoplasmic signal and control for tissue autofluorescence (HPA: supported nucleoplasm localization; standard IF practice).
HPA tissue IHC evidence for IPMK
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — 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 | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IPMK IHC Tips
Use compartment-aware controls and matched processing to assess IPMK staining in paraffin sections; interpret tissue patterns cautiously (UniProt Q8NFU5; HPA tissue IHC).
Which retrieval conditions should I try first for weak IPMK staining in paraffin sections?
Start with heat-mediated Tris-EDTA at pH 9.0 for IPMK staining in paraffin sections (datasheet A06955-1). The selected colon cancer image used that retrieval with primary antibody at 1:200, incubated at 4°C overnight; its caption does not state the fixative (caption A06955-1). If staining is weak, vary heating time within your validated instrument workflow and compare sections processed together, checking tissue preservation as well as signal (standard IHC practice). Include a section processed without primary antibody so stronger chromogen deposition after retrieval is not mistaken for specific IPMK signal (standard IHC practice).
Can fixation explain inconsistent IPMK staining across paraffin blocks?
Target-specific sensitivity of IPMK to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not report a fixative (caption A06955-1). Record each block’s fixative and fixation duration, then compare sections using identical Tris-EDTA pH 9.0 retrieval, antibody incubation and detection conditions (datasheet A06955-1; standard IHC practice). If fixation histories differ, test matched control sections alongside each batch before attributing intensity changes to biology (standard IHC practice). Do not infer a fixation effect from IPMK’s nuclear annotation, its 0 annotated glycosylation sites or the HPA tissue pattern (UniProt Q8NFU5; HPA tissue IHC).
How should I assess cytoplasmic staining when IPMK is annotated as nuclear?
Score nuclear and cytoplasmic chromogen separately: UniProt places IPMK in the nucleus, while HPA tissue IHC describes cytoplasmic expression in most tissues (UniProt Q8NFU5; HPA tissue IHC). HPA’s ICC/IF data support nucleoplasmic localization and also report basal-body localization, but those findings do not establish the compartment in every paraffin tissue section (HPA subcellular). Examine intact cells away from folds and section edges, and compare the pattern with a no-primary control (standard IHC practice). Because HPA rates its tissue IHC evidence Uncertain pending external verification, treat any single-compartment call as provisional until controls and independent staining agree (HPA tissue IHC; standard IHC practice).
Could an isoform or hidden epitope explain uneven IPMK staining?
The supplied record lists 0 isoforms and no annotated domains, so an isoform-specific interpretation is unsupported here (UniProt Q8NFU5). IPMK has no annotated transmembrane segment, signal peptide or propeptide; its annotated chain spans residues 2–416 (UniProt Q8NFU5). N-terminal acetylation at residue 2 and phosphorylation at residue 7 are recorded, but the selected antibody’s epitope is not supplied, so their effect on binding is unknown (UniProt Q8NFU5; caption A06955-1). If staining is uneven, first compare retrieval and section quality, then seek epitope documentation or an independently validated antibody before assigning the pattern to molecular processing (standard IHC practice).
How can IF help resolve ambiguous IPMK staining in tissue sections?
Use IF as a separate validation experiment, multiplexing IPMK with a marker for the cell population being assessed and a nuclear counterstain (standard IF practice). Choose spectrally separated fluorophores and place the weaker signal in a channel with low tissue autofluorescence; include single-stain and no-primary controls (standard IF practice). IPMK has no annotated transmembrane segment, and HPA supports a nucleoplasmic location, so use a validated permeabilisation step that permits antibody access to intracellular epitopes without damaging cell morphology (UniProt Q8NFU5; HPA subcellular; standard IF practice). Compare compartments with chromogenic IHC, but do not transfer the paraffin-section retrieval or 1:200 dilution directly to IF (caption A06955-1; standard IF practice).
What should I change when IPMK chromogen appears diffuse or widespread?
Run a no-primary section and inspect whether pigment persists, especially around vessels, damaged areas and tissue edges (standard IHC practice). For peroxidase-based chromogenic detection, verify the peroxidase block, wash stringency and development time; these are general workflow controls, not evidence of IPMK-specific behavior (standard IHC practice). The selected paraffin image used primary antibody at 1:200 overnight at 4°C and secondary antibody at 1:200 for 45 minutes at room temperature, providing a documented starting point for that SKU (caption A06955-1). If background remains high, titrate the antibody and shorten chromogen development while preserving matched controls (standard IHC practice).
How should I quantify IPMK staining when nuclear and cytoplasmic signals differ? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before measuring sections; report nuclear and cytoplasmic results separately because UniProt and HPA describe differing localization evidence (UniProt Q8NFU5; HPA tissue IHC). For cellular staining, use an H-score or percentage of positive cells with fixed intensity thresholds across the study (standard IHC practice). If counting positive cells by area, report density per mm² of viable, annotated tissue and normalize to the number of eligible cells or tissue area, as appropriate (standard IHC practice). Exclude folds, edges and necrotic regions consistently, and compare sections with matched retrieval, staining and imaging settings (standard IHC practice).
What evidence distinguishes genuine IPMK staining from an artefact?
A credible result should localize to intact cells, recur across comparable sections and exceed the no-primary signal under matched conditions (standard IHC practice). Interpret compartment carefully: UniProt annotates nuclear IPMK, HPA ICC/IF supports nucleoplasm, and HPA tissue IHC describes mostly cytoplasmic staining with Uncertain reliability (UniProt Q8NFU5; HPA subcellular; HPA tissue IHC). Check whether the stained cell type fits the specific tissue observation; for example, HPA reports medium staining in colon endothelial cells, not a blanket positive call for every colon cell (HPA tissue IHC). Discount edge effects, necrosis and residual endogenous peroxidase signal before drawing a biological conclusion from DAB intensity (standard IHC practice).
Boster reagents
Best IPMK / Inositol polyphosphate multikinase IHC Antibodies
A06955-1 has a human paraffin-section IHC image (catalog image caption). IF is listed, but no IF image is provided (catalog applications; catalog IF images). Human, monkey, mouse and rat reactivity is listed (catalog reactivity).
Real IHC data
Anti-IPMK Antibody
Cat # A06955-1
A06955-1 will render with an IHC image from paraffin-embedded human colon cancer (catalog image caption). The same SKU lists IF among its applications and human, monkey, mouse and rat reactivity, but provides no IF image (catalog applications; catalog reactivity; catalog IF images).
Which to pick: For tissue IHC, choose A06955-1: its own image documents paraffin-embedded human colon cancer, 1:200 primary antibody overnight at 4°C, and Tris-EDTA retrieval at pH 9.0; the fixative is unreported (catalog image caption). For IF, A06955-1 lists a 1:50 dilution, but no IF image or ICC validation is provided (catalog IF dilution; catalog IF images; catalog applications). For cross-species work, A06955-1 is a polyclonal antibody with human, monkey, mouse and rat reactivity listed; its supplied IHC image documents human tissue only (catalog dilution data; catalog reactivity; catalog image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NFU5 (IPMK_HUMAN, Inositol polyphosphate multikinase).
- Human Protein Atlas. IPMK tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. IPMK subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the basal body..
- Human Protein Atlas. IPMK antibody validation summary (2 antibodies).
- Ebastine exerts antitumor activity and induces autophagy by activating AMPK/ULK1 signaling in an IPMK-dependent manner in osteosarcoma. International journal of biological sciences 2023 — PMC9830506.
- Immunotherapy landscape analyses of necroptosis characteristics for breast cancer patients. Journal of translational medicine 2022 — PMC9306193.
- Comprehensive Analysis of Necroptosis-Related Genes as Prognostic Factors and Immunological Biomarkers in Breast Cancer. Journal of personalized medicine 2022 — PMC9863352.
- Loss of PI3k activity of inositol polyphosphate multikinase impairs PDK1-mediated AKT activation, cell migration, and intestinal homeostasis. iScience 2023 — PMC10197106.
- PubMed PMID:12027805 — UniProt-cited evidence.
- PubMed PMID:12223481 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IPMK (Inositol polyphosphate multikinase). Expected localisation Cytoplasmic in tissue (HPA tissue IHC); nuclear molecularly (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A06955-1). Validated IHC antibody: A06955-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Colon cancer tissue; fixative not specified (datasheet A06955-1); 2. Fixation: Image fixative and duration unreported (datasheet A06955-1); 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 retrieval: Tris-EDTA pH 9.0 (datasheet A06955-1); 20 min, 95–100 °C (standard); 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-IPMK, 1:100-1:300 (datasheet A06955-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Tissue staining awaits external verification (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one chain spans residues 2–416 (UniProt); Regulation: Highest expression in skeletal muscle and liver (UniProt). Evidence: UniProt Q8NFU5, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A06955-1, PMC10197106. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IPMK/immunohistochemistry-ihc-guide.