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IP6K1 / Inositol hexakisphosphate kinase 1 · IHC design guide
Design Immunohistochemistry for IP6K1
Plan chromogenic IP6K1 IHC on paraffin sections using the IHC-validated antibody at 1:50–1:200 (datasheet A07884). Assess cytoplasmic and nuclear staining in glandular cells while accounting for low consistency between antibody staining and RNA (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IP6K1 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 and nuclear staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Glandular cells show cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A07884) | |
| 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 | Antibody staining and RNA show low consistency (HPA tissue IHC) | |
| Regulation | Low tissue RNA specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended IP6K1 IHC & IF Protocols
Compare the catalog antibody’s citrate pH 6.0 IHC-P protocol (datasheet: A07884) with a published breast tumor TMA protocol (PMC7976380).
| Sample | Paraffin-embedded rat kidney tissue; fixative not specified (datasheet A07884) |
| Fixation | Image fixative and duration unreported (datasheet A07884); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A07884); 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-IP6K1, 1:50-1:200 (datasheet A07884) |
| 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 | IP6K1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval for the catalog antibody (datasheet: A07884). Use the published low-pH schedule when reproducing the TMA protocol (PMC7976380).
Section 2
What Is the Expected IP6K1 Staining Pattern?
IP6K1 is found in the cytoplasm and nucleus (UniProt Q92551; HPA tissue IHC). In paraffin sections, expect staining in many tissues, including strong staining of glandular cells in several organs and skeletal muscle myocytes (HPA tissue IHC). IP6K1 has no annotated transmembrane segment (UniProt Q92551 topology). HPA rates its tissue staining Approved, while noting low agreement with RNA data and pending external verification (HPA tissue IHC).
| Cytoplasmic and nuclear staining in glandular cells of colon or duodenum. | This fits the reported high glandular-cell staining and broad compartment pattern (HPA tissue IHC). Compare cells within the section; the HPA rating is Approved, with low agreement between staining and RNA data (HPA tissue IHC). |
| Strong staining in skeletal muscle myocytes. | High myocyte staining is reported (HPA tissue IHC). Judge the cellular pattern alongside morphology and the negative control; a strong chromogen deposit alone does not establish antibody specificity (general IHC practice). |
| Staining confined to cell membranes, with no cytoplasmic or nuclear signal. | A membrane-only pattern conflicts with the reported locations (HPA tissue IHC) and the lack of a transmembrane segment (UniProt Q92551 topology). Treat it as suspect and check detection background and tissue morphology (general IHC practice). |
| Strong staining in adipocytes or lymph-node germinal-center cells. | These cell types are reported as not detected (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity before calling them IP6K1-positive; confirm the pattern with controls and independent evidence (general IHC practice). |
| Haze across cells, stroma, and blank areas of the section. | Diffuse staining that ignores cell boundaries cannot establish the reported cytoplasmic and nuclear pattern (HPA tissue IHC). Review the negative control and detection steps for nonspecific or endogenous signal (general IHC practice). |
💡Expected IP6K1 appearanceCall a section positive when identifiable glandular cells in a reported high-staining tissue show strong cytoplasmic and nuclear staining (HPA tissue IHC); membrane-only deposits or diffuse staining outside cell boundaries are suspect (UniProt Q92551 topology; general IHC practice).
| Cellular compartment and topology | Cytoplasm and nucleus are the reported locations (UniProt Q92551; HPA tissue IHC). No transmembrane segment is annotated (UniProt Q92551 topology), so membrane-only staining needs investigation; neither source makes membrane staining a validated readout. |
| Choice of reference cells | Glandular cells in colon, duodenum, and appendix, and skeletal muscle myocytes, are reported High (HPA tissue IHC). Adipocytes and lymph-node germinal-center cells are reported Not detected (HPA tissue IHC); use those calls at the named cell-type level. |
| Antibody evidence | HPA040825 is listed as IHC Approved and ICC Supported (HPA antibodies). The tissue profile carries a warning about low agreement with RNA expression and awaits external verification (HPA tissue IHC); a matching image is supportive, not conclusive. |
| Isoforms and processing | Two isoforms are listed, and the annotated protein chain spans residues 1–441, with no signal peptide or propeptide (UniProt Q92551). The payload gives no antibody epitope, so it cannot establish whether both isoforms stain equally. |
| IF/ICC Q: What localisation should I expect? | A: HPA reports a supported main location at the nucleoli fibrillar center, with supported nucleoplasm and cytosol locations in ICC-IF (HPA subcellular). This higher-resolution observation does not require nucleoli to be resolved in chromogenic tissue IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in colon or duodenal glandular cells. | These cells are reported High (HPA tissue IHC); a failed assay or unsuitable section remains possible (general IHC practice). | Check section integrity, primary-antibody and detection steps, and a concurrently stained reference section. Optimize retrieval or dilution only against the actual antibody instructions; no IP6K1-specific retrieval setting is supplied. |
| All cells stain uniformly, including HPA-reported negative cell types. | Adipocytes and lymph-node germinal-center cells are reported Not detected (HPA tissue IHC); uniform signal may reflect background or endogenous detection activity (general IHC practice). | Inspect a matched negative control, blocking, washes, and chromogen development. Reassess staining by identifiable cell type after background is controlled (general IHC practice). |
| Only a membrane outline is visible. | The pattern conflicts with cytoplasmic and nuclear localisation (HPA tissue IHC) and lacks support from the annotated topology (UniProt Q92551). | Check whether outlines also appear in the negative control or along section edges. Repeat with controlled detection conditions and assess cytoplasm and nucleus (general IHC practice). |
| Nuclear staining is absent, but cytoplasm stains. | HPA describes cytoplasmic and nuclear expression in most tissues (HPA tissue IHC); the balance in an individual section is not specified. | Confirm that the counterstain and morphology permit nuclear assessment, then compare a reported positive cell type and controls. Do not force a nuclear-positive call from cytoplasmic signal alone (general IHC practice). |
| Staining appears in a cell type HPA calls Not detected. | HPA's call is cell-type specific and its tissue IHC profile has low agreement with RNA data (HPA tissue IHC); cross-reactivity or endogenous activity also remains possible (general IHC practice). | Verify cell identity and localisation, inspect the negative control, and seek independent antibody or orthogonal evidence before assigning IP6K1 expression (general IHC practice). |
| IF/ICC shows punctate nuclear signal that differs from the IHC image. | HPA ICC-IF places IP6K1 mainly at the nucleoli fibrillar center, with nucleoplasm and cytosol also supported (HPA subcellular). Chromogenic tissue IHC has different spatial resolution (general microscopy practice). | Interpret each assay at its own resolution; compare nuclear and cytoplasmic distribution in IHC without requiring visible nucleolar puncta. Use the separate IF/ICC guide for that assay's procedure. |
Experiment evidence
IHC & IF images showing IP6K1 staining
A07884 · IHCImmunohistochemistry of paraffin-embedded rat kidney using IP6K1 Rabbit pAb at dilution of 1:100 .Perform high pressure antigen retrieval wi
A07884 · IHCImmunohistochemistry of paraffin-embedded mouse kidney using IP6K1 Rabbit pAb at dilution of 1:100 .Perform high pressure antigen retrieval
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000176095-IP6K1/tissue/cerebral+cortex#imid_10441117
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000176095-IP6K1/tissue/colon#imid_10441027
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000176095-IP6K1/tissue/liver#imid_10441036
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000176095-IP6K1/tissue/kidney#imid_10441045
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA040825; green = IP6K1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA040825; green = IP6K1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IP6K1 IHC & IF
🧪Run colon first and look for staining in glandular cells (HPA: High in colon glandular cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); on the colon slide, assess adjacent nonglandular areas for background without assuming those cells are IP6K1-negative (HPA: High is reported for glandular cells).
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 IP6K1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoli fibrillar center (supported) (HPA subcellular).
Technical controls: Include a no-primary/secondary-only control, matched normal rabbit IgG for the rabbit polyclonal catalog antibody, and IP6K1-knockout material if available (A07884 caption: rabbit pAb; standard IHC practice). Quench endogenous peroxidase before chromogenic detection and check colon mucus and inflammatory areas for nonspecific signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A07884 paraffin-section caption does not state the fixative (A07884 caption: fixative unreported). Retrieval dependency is untested: the caption specifies high-pressure retrieval in 10 mM citrate at pH 6.0 before rat kidney IHC, but provides no comparison without retrieval (A07884 caption). Whether frozen sections or IF are easier is unreported; colon tissue autofluorescence and mucus can complicate IF interpretation (standard IF practice).
HPA tissue IHC evidence for IP6K1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IP6K1 IHC Tips
Use compartment-aware controls and consistent section processing to troubleshoot IP6K1 staining in paraffin sections (UniProt Q92551; HPA tissue IHC).
How should I adjust retrieval when IP6K1 staining is weak in paraffin sections?
Start with heat-mediated retrieval in 10 mM citrate buffer at pH 6.0 (datasheet A07884). The paraffin-section image used high-pressure retrieval before staining at 1:100, although its caption does not report fixation (caption A07884). If signal remains weak, compare a shorter and longer heating interval on adjacent sections while keeping antibody dilution, detection, and counterstain constant (general IHC practice). Check a no-primary control for retrieval-related background and inspect tissue structure for damage (general IHC practice). Score nuclear and cytoplasmic signal separately because both compartments are expected for IP6K1 (UniProt Q92551 localization; HPA tissue IHC).
Could fixation explain weak or uneven IP6K1 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A07884). Record the fixative, fixation duration, section thickness, and time in storage for each specimen before comparing staining (general IHC practice). Process matched sections with the same citrate retrieval at pH 6.0 and the same antibody dilution to isolate fixation as a possible variable (datasheet A07884; general IHC practice). Compare preserved cell morphology and staining in an internal positive compartment rather than judging intensity alone (general IHC practice). A negative result after one fixation condition does not establish absence of IP6K1 without a suitable positive control (general IHC practice).
Is predominantly nuclear or cytoplasmic IP6K1 staining credible?
Both nuclear and cytoplasmic staining can be credible for IP6K1 (UniProt Q92551 localization; HPA tissue IHC profile). Evaluate those compartments within intact cells and record their intensities separately, using the counterstain to define nuclei (general IHC practice). The subcellular IF profile places signal mainly in the nucleoli fibrillar center, with additional nucleoplasmic and cytosolic signal; that finer pattern should not be assumed resolvable by chromogenic tissue IHC (HPA subcellular). If staining instead outlines the plasma membrane throughout the section, investigate nonspecific detection because IP6K1 has no annotated transmembrane segment (UniProt Q92551 topology). Recheck the pattern against a no-primary control and adjacent tissue morphology (general IHC practice).
Can epitope choice change which IP6K1 signal I detect?
IP6K1 has 2 annotated isoforms, so epitope coverage matters when comparing antibodies or interpreting discordant sections (UniProt Q92551 isoforms). Check the antibody's stated immunogen or epitope against both isoform sequences before attributing a staining difference to biology (general IHC practice). The supplied record lists a phosphoserine at residue 151, but it does not establish whether the catalog antibody recognizes a phosphorylation-dependent epitope (UniProt Q92551 modified residues; supplied antibody evidence). Keep retrieval and detection identical across antibody comparisons, and assess nuclear and cytoplasmic staining independently (datasheet A07884; UniProt Q92551 localization; general IHC practice). Treat isoform-specific claims as unresolved until the epitope is mapped (general IHC practice).
How can I check IP6K1 localisation by multiplex IF?
Use a separate IF/ICC-validated setup to assess subcellular detail; the selected product evidence describes paraffin-section IHC (caption A07884). Multiplex IP6K1 with a marker for the cell type being evaluated and a nuclear stain, then inspect single channels before merging them (general IF practice). Choose spectrally separated fluorophores and place the weaker signal in a channel with low measured tissue autofluorescence (general IF practice). For intracellular nuclear and cytosolic epitopes, use a mild permeabilisation condition and compare it with an unpermeabilised control; IP6K1 has no annotated transmembrane segment (UniProt Q92551 topology and localization; general IF practice). The reported nucleolar, nucleoplasmic, and cytosolic pattern provides a compartment-level comparison (HPA subcellular).
What should I change if IP6K1 staining is diffuse or widespread?
Run a no-primary control to distinguish antibody-associated staining from detection chemistry and tissue pigment (general chromogenic IHC practice). For peroxidase detection, include an endogenous peroxidase block and compare a DAB-only control where background is suspected (general chromogenic IHC practice). Titrate the primary antibody around the caption's 1:100 condition on matched sections, holding citrate retrieval at pH 6.0 constant (caption A07884; datasheet A07884; general IHC practice). Reduce excessive chromogen development or adjust blocking and washing if control sections show diffuse deposit (general IHC practice). Judge residual staining by cell and compartment: widespread cytoplasmic and nuclear expression is reported, but uniform extracellular color needs investigation (HPA tissue IHC; general IHC practice).
How should I quantify IP6K1 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and tissue area before viewing the stain, then exclude folds, edges, and necrotic regions by the same rule for every slide (general IHC practice). For nuclear and cytoplasmic IP6K1, report separate percentages of positive cells or separate H-scores rather than combining compartments (UniProt Q92551 localization; HPA tissue IHC; general IHC practice). An H-score sums percentages at intensity grades 0–3, yielding 0–300 for each compartment (general IHC practice). Normalize counts to evaluable cells, or positive-cell density to analyzed area in mm², and use matched acquisition and threshold settings (general IHC practice). Include a consistent reference section across staining runs (general IHC practice).
Which patterns support genuine IP6K1 staining rather than artefact?
Give greatest weight to intracellular nuclear or cytoplasmic staining in intact cells, the reported IP6K1 compartments (UniProt Q92551 localization; HPA tissue IHC). Glandular cells in colon and adrenal gland are reported as high, while adipocytes in adipose tissue are reported as not detected; use such cell-level patterns as context, not absolute controls (HPA tissue IHC). Investigate staining confined to section edges, necrotic areas, or no-primary controls as possible processing or detection artefact (general IHC practice). Check a membrane-only pattern against the absence of an annotated transmembrane segment, and check brown deposits against an endogenous peroxidase control (UniProt Q92551 topology; general chromogenic IHC practice). HPA notes low agreement between staining and RNA data and pending external verification, so corroborate consequential conclusions independently (HPA tissue IHC reliability).
Boster reagents
Best IP6K1 / Inositol hexakisphosphate kinase 1 IHC Antibodies
A07884 has real IHC images from paraffin-embedded rat and mouse kidney; human reactivity is listed but not pictured (A07884 IHC captions; catalog: reactivity).
Real IHC data
Anti-IP6K1 Antibody
Cat # A07884
A07884 is listed for IHC and has images of paraffin-embedded rat and mouse kidney stained at 1:100 (catalog: applications; A07884 IHC captions). The catalog also lists human reactivity, but provides no human IHC image or IF image (catalog: reactivity and image alts).
Which to pick: For tissue IHC, choose A07884, a rabbit polyclonal antibody listed for IHC at 1:50–1:200; its captions document citrate pH 6.0 retrieval in paraffin-embedded rat and mouse kidney (catalog: host and IHC dilution; A07884 IHC captions). For cross-species work, A07884 lists human, mouse and rat reactivity, although only mouse and rat IHC images are supplied; the fixative is unreported (catalog: reactivity; A07884 IHC captions). No SKU in this payload is listed for IF/ICC (catalog: A07884 applications).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92551 (IP6K1_HUMAN, Inositol hexakisphosphate kinase 1).
- Human Protein Atlas. IP6K1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. IP6K1 subcellular location (ICC-IF): Mainly localized to the nucleoli fibrillar center. In addition localized to the nucleoplasm and cytosol..
- Human Protein Atlas. IP6K1 antibody validation summary (1 antibodies).
- IP6K2 predicts favorable clinical outcome of primary breast cancer. Molecular and clinical oncology 2021 — PMC7976380.
- PubMed PMID:9039502 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16641997 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IP6K1 (Inositol hexakisphosphate kinase 1). Expected localisation Cytoplasmic and nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A07884). Validated IHC antibody: A07884 at 1:50-1:200 (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 rat kidney tissue; fixative not specified (datasheet A07884); 2. Fixation: Image fixative and duration unreported (datasheet A07884); 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: Citrate pH 6.0 (datasheet A07884); 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-IP6K1, 1:50-1:200 (datasheet A07884); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show low consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: Low tissue RNA specificity (HPA tissue RNA). Evidence: UniProt Q92551, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoli fibrillar center. In addition…, Datasheet A07884, PMC7976380. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IP6K1/immunohistochemistry-ihc-guide.