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IMPDH1 / Inosine-5'-monophosphate dehydrogenase 1 · IHC design guide
Design Immunohistochemistry for IMPDH1
Plan chromogenic IMPDH1 IHC in paraffin sections using cytoplasmic and nuclear staining as the tissue reference (HPA tissue IHC). Start with the catalog antibody’s 2–5 μg/ml IHC range (datasheet A03791-1), then score staining by cell type and compartment.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IMPDH1 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 (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear staining; high in spermatogonia (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03791-1) | |
| Positive control | Testis+4 more · see all | |
| Negative control | Skeletal muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining may include products of another gene (HPA tissue IHC) | |
| Regulation | Type II predominates in tumors (UniProt) | |
| Isoform / epitope | 7 isoforms; check epitope coverage across them (UniProt) |
Section 1
Recommended IMPDH1 IHC & IF Protocols
The catalog antibody protocol (datasheet: A03791-1) is followed by 4 published IMPDH1 IHC protocols (PMC9945078; PMC10197894; PMC9102537; PMC9733023).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A03791-1) |
| Fixation | Image fixative and duration unreported (datasheet A03791-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A03791-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03791-1) |
| Primary antibody | Rabbit anti-IMPDH1, 2-5 μg/ml (datasheet A03791-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03791-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03791-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | IMPDH1-positive staining in spermatogonia cells of testis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 (datasheet: A03791-1); published alternatives use Tris-EDTA at pH 9.0 or citrate-based retrieval (PMC9945078; PMC9102537; PMC9733023).
Section 2
What Is the Expected IMPDH1 Staining Pattern?
IMPDH1 is a cytoplasmic and nuclear protein with no transmembrane segment (UniProt P20839). In tissue IHC, expect cytoplasmic and nuclear staining in several cell types, including high staining in testis spermatogonia (HPA: tissue IHC). HPA rates the tissue profile Approved with medium consistency against RNA data and cautions that the antibody targets proteins from more than one gene (HPA: tissue IHC reliability).
| Distinct cytoplasmic and nuclear staining in spermatogonia, with stronger signal than nearby background. | This fits the reported high spermatogonial staining and the broader cytoplasmic and nuclear tissue profile (HPA: testis IHC; HPA: tissue IHC profile). Score the stained cell type and compartment separately. The pattern supports IMPDH1-associated staining, but HPA's multi-gene targeting caution limits attribution to IMPDH1 alone (HPA: tissue IHC reliability). |
| Staining appears predominantly at cell borders, with little cytoplasmic or nuclear signal. | A border-restricted pattern is unexpected for a protein reported in the cytoplasm and nucleus without a transmembrane segment (UniProt P20839; HPA: tissue IHC profile). Treat it as a possible staining artefact and review tissue morphology, detection controls and antibody specificity before assigning it to IMPDH1 (general IHC practice). |
| Strong staining appears in skeletal-muscle myocytes or another cell population selected as a negative comparator. | HPA reports IMPDH1 as not detected in skeletal-muscle myocytes (HPA: skeletal muscle IHC). Check whether the signal belongs to myocytes rather than adjacent cells. Staining in confirmed myocytes warrants investigation of cross-reactivity or endogenous detection activity; the HPA multi-gene antibody caution makes that check especially relevant (HPA: tissue IHC reliability; general IHC practice). |
| Chromogen covers many structures uniformly, obscuring cell borders and nuclei. | Diffuse signal without a readable cellular distribution cannot establish the reported cytoplasmic and nuclear pattern (HPA: tissue IHC profile). Compare a reagent omission control and inspect section edges and tissue folds; uneven reagent deposition or nonspecific detection can produce background in chromogenic IHC (general IHC practice). |
| No staining is visible in testis spermatogonia on a run intended to show a positive control. | That conflicts with HPA's high spermatogonial staining and suggests a run or specimen problem before it supports a biological absence call (HPA: testis IHC; general IHC practice). Confirm that spermatogonia are present, then review the antibody dilution, retrieval, detection reagents and control slide together (general IHC practice). |
💡Expected IMPDH1 appearanceA convincing IHC positive shows readable cytoplasmic and nuclear staining in spermatogonia, where HPA reports high intensity, while isolated border staining or chromogen across cell-free areas calls for artefact checks (HPA: testis IHC; HPA: tissue IHC profile; UniProt P20839; general IHC practice).
| Compartment and topology | Cytoplasm and nucleus are reported locations, and no transmembrane segment is listed (UniProt P20839). Use compartment as an interpretation check; topology alone does not predict which compartment will dominate on a given slide. |
| Cell-specific reference levels | HPA reports high staining in spermatogonia, medium staining in several other listed cell types, and no detection in skeletal-muscle myocytes (HPA: tissue IHC). These are reference observations, not guaranteed results for every specimen. |
| Antibody evidence and specificity | The HPA tissue IHC profile is Approved with medium RNA concordance; its antibody may target proteins from more than one gene (HPA: tissue IHC reliability). A matching pattern does not by itself prove that every stained cell contains IMPDH1. |
| Isoform coverage | Seven IMPDH1 isoforms are listed (UniProt P20839). Their presence does not establish which isoforms an antibody recognizes; assess coverage only when an epitope or validation record is available. |
| IF appearance | ICC-IF reports supported cytosol and approved rods and rings, with the same multi-gene antibody caution (HPA: subcellular ICC-IF). Treat this as context for an IF image, not as an expected chromogenic tissue IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive-control testis section has no spermatogonial signal. | The run may have failed, or the relevant cells may be absent from the examined section (HPA: high in spermatogonia; general IHC practice). | Verify cell identity and morphology; compare a previously working positive section and review the recorded retrieval, antibody dilution and detection steps (general IHC practice). Do not score the specimen negative until the control works. |
| Only nuclei stain in cells expected to show a mixed pattern. | Nuclear localisation is plausible, but tissue IHC reports cytoplasmic and nuclear expression; a nucleus-only result needs review (UniProt P20839; HPA: tissue IHC profile). | Inspect whether weak cytoplasmic signal is hidden by counterstain or background, then compare compartment patterns in a known-positive section (general IHC practice). Avoid calling nuclear staining alone proof of specificity. |
| Staining outlines cells with little internal signal. | A border-dominant result does not match the reported cytoplasmic and nuclear distribution or the absence of a transmembrane segment (HPA: tissue IHC profile; UniProt P20839). | Examine morphology and a reagent omission control for edge deposits or nonspecific detection; verify the pattern with independent specificity evidence if available (general IHC practice). |
| Skeletal-muscle myocytes stain strongly. | HPA reports no detection in myocytes; signal could reflect misidentified cells, cross-reactivity or endogenous detection activity (HPA: skeletal muscle IHC; HPA: multi-gene antibody caution; general IHC practice). | Confirm that the stained cells are myocytes, review the detection control, and compare an independent antibody or orthogonal evidence where available (general IHC practice). |
| Diffuse chromogen prevents cell-level scoring. | Background can arise from nonspecific reagent binding or endogenous detection activity in chromogenic IHC (general IHC practice). It cannot be interpreted as the HPA cytoplasmic and nuclear pattern (HPA: tissue IHC profile). | Use a reagent omission control to locate the background source; review blocking, washing and detection exposure, then score only a section with clear cell boundaries (general IHC practice). |
| IF/ICC shows rods and rings; should they be called an IHC tissue positive? | HPA reports rods and rings in ICC-IF alongside cytosol, with a warning that the antibodies can target proteins from multiple genes (HPA: subcellular ICC-IF). | Interpret rods and rings within the separate IF/ICC context. For paraffin tissue IHC, use the reported cytoplasmic and nuclear tissue pattern and cell-specific reference levels; do not transfer the ICC-IF pattern into an IHC scoring rule (HPA: tissue IHC; HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing IMPDH1 staining
A03791-1 · IHCIHC analysis of IMPDH1 using anti-IMPDH1 antibody (A03791-1). IMPDH1 was detected in a paraffin-embedded section of human breast cancer tiss
A03791-1 · IHCIHC analysis of IMPDH1 using anti-IMPDH1 antibody (A03791-1). IMPDH1 was detected in a paraffin-embedded section of human lymphoma tissue. H
A03791-1 · IFIF analysis of IMPDH1 using anti-IMPDH1 antibody (A03791-1). IMPDH1 was detected in an immunocytochemical section of PC-3 cells. Enzyme anti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000106348-IMPDH1/tissue/cerebral+cortex#imid_665040
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000106348-IMPDH1/tissue/colon#imid_664966
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000106348-IMPDH1/tissue/liver#imid_664967
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000106348-IMPDH1/tissue/kidney#imid_664984
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001400; green = IMPDH1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001400; green = IMPDH1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IMPDH1 IHC & IF
🧪Run testis first and assess staining in spermatogonia (High; HPA: testis, spermatogonia). Use skeletal muscle myocytes as the negative tissue (HPA: skeletal muscle, not detected); compare other cells on the testis slide for background, but do not assume they are negative because their staining is unreported (HPA: testis row).
Positive control tissue: Testis (Spermatogonia cells, HPA High)
Negative control tissue: Skeletal muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show IMPDH1 in A-431, U-251MG, U2OS, ASC52telo, hTERT-RPE1 (serum starved), with annotated localisation: Cytosol (supported), Rods & Rings (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and rabbit IgG isotype controls matched to the primary antibody’s clonality, plus an IMPDH1-knockout specimen if available (caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase in testis sections before HRP/DAB detection (caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: An IMPDH1-specific fixation window or fixation effect is unreported, and the selected A03791-1 tissue-IHC caption does not state a fixative (selected-SKU caption). That paraffin-section example used heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required for every specimen (selected-SKU caption). The supplied evidence does not establish that frozen sections or IF are easier, or identify a testis-specific artefact (selected-SKU caption; HPA: testis row).
HPA tissue IHC evidence for IMPDH1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IMPDH1 IHC Tips
Troubleshoot IMPDH1 staining in paraffin sections by checking retrieval, compartment, cell type, and the limits of antibody specificity.
How should I troubleshoot weak IMPDH1 staining after antigen retrieval?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A03791-1). The selected tissue image used this retrieval before overnight incubation with 2 μg/ml catalog antibody at 4°C, but its caption does not state the fixative (datasheet A03791-1). If staining remains weak, compare retrieval duration on adjacent sections while keeping antibody concentration and detection constant; monitor tissue damage and nonspecific staining (standard IHC practice). Score expected cytoplasmic and possible nuclear staining in the same cell types across conditions, since IMPDH1 is reported in both compartments (UniProt P20839; HPA tissue IHC).
Could fixation explain a weak or uneven IMPDH1 IHC signal?
Target-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not report a fixative (datasheet A03791-1). Record the specimen’s fixative, fixation interval, processing history, and section age before comparing staining across cases, because these variables can affect antigen preservation and retrieval (standard IHC practice). Use adjacent sections to assess morphology and repeat the documented EDTA pH 8.0 retrieval with matched detection conditions (datasheet A03791-1; standard IHC practice). If signal differs between specimens, report that difference alongside processing details rather than assigning it to an IMPDH1-specific fixation effect (standard IHC practice).
Which staining compartments are plausible for IMPDH1 in tissue sections?
Cytoplasmic and nuclear staining are plausible for IMPDH1 (UniProt P20839; HPA tissue IHC). Its sequence has no transmembrane segment, so a crisp membrane-only pattern warrants scrutiny rather than assignment to the target (UniProt P20839 topology; standard IHC interpretation). Assess cytoplasmic and nuclear signal separately within morphologically identified cells, using the counterstain to define nuclei (standard IHC practice). HPA reports cytosol and rods and rings in subcellular imaging, but that observation comes with a warning that the antibodies target proteins from multiple genes; do not require visible rods and rings in chromogenic tissue IHC (HPA subcellular).
How can IMPDH1 isoforms or epitope accessibility complicate staining?
IMPDH1 has 7 listed isoforms and two CBS domains at residues 114–173 and 179–237 (UniProt P20839). The supplied caption does not identify the catalog antibody’s epitope, so its coverage of individual isoforms cannot be established here (datasheet A03791-1). Compare staining with an independently characterized antibody whose epitope is known, and keep retrieval and detection conditions matched when investigating discordance (standard IHC practice). Interpret differences cautiously: phosphorylation at residue 160 and methylation at residues 341 and 355 are recorded, but no supplied evidence shows that these modifications alter this antibody’s staining (UniProt P20839).
How should I assess an IMPDH1 signal in multiplex immunofluorescence?
For an IF comparison, pair IMPDH1 with a marker identifying the expected cell population and inspect each channel independently before interpreting overlap (standard IF practice). HPA reports high staining in spermatogonia and medium staining in adipocytes, with a caution that its tissue antibody can recognize proteins from more than one gene (HPA tissue IHC). Choose fluorophores and imaging channels around the specimen’s measured autofluorescence, then confirm that apparent colocalisation survives single-channel inspection (standard IF practice). IMPDH1 has no transmembrane segment and is reported in cytoplasm and nucleus, so choose permeabilisation that allows antibody access to those compartments and verify its effect empirically (UniProt P20839; standard IF practice).
What should I check when IMPDH1 DAB staining appears diffuse?
The selected paraffin-section image used 10% goat serum blocking, a peroxidase-conjugated secondary antibody, and DAB development (datasheet A03791-1). Check a no-primary control for secondary-antibody binding and endogenous peroxidase signal, and use a peroxidase block before chromogen development as a general IHC step (standard IHC practice). Compare background in tissue edges, damaged regions, and cell-free spaces with signal inside intact cells; adjust washing and detection exposure if background obscures cellular boundaries (standard IHC practice). Retain the documented 2 μg/ml primary concentration as the reference condition when testing background controls, changing one variable at a time (datasheet A03791-1; standard IHC practice).
How should I quantify IMPDH1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring, then record cytoplasmic and nuclear staining separately because both locations are reported for IMPDH1 (UniProt P20839; HPA tissue IHC; standard IHC practice). Within each compartment, use percent positive cells or an H-score combining positive fraction and intensity; density per mm² is useful when cell abundance itself is the outcome (standard IHC practice). Normalise cell-based scores to the number of eligible, intact cells and area-based counts to evaluable tissue area, excluding folds and necrosis by a prespecified rule (standard IHC practice). Apply identical illumination, threshold, counterstain review, and scoring rules across slides (standard IHC practice).
When is an IMPDH1-positive IHC pattern convincing rather than artefactual?
A convincing result places signal within intact cells and in plausible cytoplasmic or nuclear compartments (UniProt P20839; HPA tissue IHC; standard IHC interpretation). Compare cell identity and staining distribution with controls: HPA reports high staining in spermatogonia, while skeletal-muscle myocytes were not detected, but its tissue assessment warns of recognition of proteins from more than one gene (HPA tissue IHC). Treat membrane-only signal, tissue-edge concentration, necrotic debris, and staining retained in a no-primary control as reasons to investigate artefact or endogenous enzyme activity (UniProt P20839 topology; standard IHC practice). Interpret tumor staining cautiously because IMPDH type II can predominate over type I in tumors (UniProt P20839).
Boster reagents
Best IMPDH1 / Inosine-5'-monophosphate dehydrogenase 1 IHC Antibodies
A03791-1 has IHC data from human paraffin sections and IF data from PC-3 cells (catalog image captions). Its listed reactivity covers Human and Mouse (datasheet).
Real IHC data
Anti-IMPDH1 Antibody ®
Cat # A03791-1
A03791-1 was demonstrated by IHC on paraffin sections of human breast cancer, lymphoma, renal cell carcinoma and spleen rupture tissue (catalog IHC image captions). A03791-1 was demonstrated by IF in PC-3 cells (catalog IF image caption).
Which to pick: Choose A03791-1 for human paraffin-section IHC: its captions report EDTA retrieval at pH 8.0, 2 μg/ml primary antibody and HRP/DAB detection; the fixative is unreported (catalog IHC image captions). For IF/ICC, A03791-1 lists both applications (datasheet) and has PC-3 cell IF data at 5 μg/ml (catalog IF image caption). For cross-species planning, A03791-1 lists Human and Mouse reactivity and a rabbit host, while its clonality is unreported (datasheet); the supplied IHC and IF images show human tissue and PC-3 cells, respectively (catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P20839 (IMDH1_HUMAN, Inosine-5'-monophosphate dehydrogenase 1).
- Human Protein Atlas. IMPDH1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. IMPDH1 subcellular location (ICC-IF): Localized to the cytosol and rods & rings. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. IMPDH1 antibody validation summary (1 antibodies).
- Potential prognostic and predictive value of UBE2N, IMPDH1, DYNC1LI1 and HRASLS2 in colorectal cancer stool specimens. Biomedical reports 2023 — PMC9945078.
- LncRNA UCA1 Participates in De Novo Synthesis of Guanine Nucleotides in Bladder Cancer by Recruiting TWIST1 to Increase IMPDH1/2. International journal of biological sciences 2023 — PMC10197894.
- Inosine monophosphate dehydrogenase type1 sustains tumor growth in hepatocellular carcinoma. Journal of clinical laboratory analysis 2022 — PMC9102537.
- MYBL2 regulates de novo purine synthesis by transcriptionally activating IMPDH1 in hepatocellular carcinoma cells. BMC cancer 2022 — PMC9733023.
- PubMed PMID:1969416 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IMPDH1 (Inosine-5'-monophosphate dehydrogenase 1). Expected localisation Cytoplasmic and nuclear staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03791-1). Validated IHC antibody: A03791-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Spermatogonia cells); negative Skeletal muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A03791-1); 2. Fixation: Image fixative and duration unreported (datasheet A03791-1); 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: EDTA pH 8.0 (datasheet A03791-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03791-1); 8. Primary antibody: Rabbit anti-IMPDH1, 2-5 μg/ml (datasheet A03791-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03791-1). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Antibody staining may include products of another gene (HPA tissue IHC); Isoform / epitope: 7 isoforms; check epitope coverage across them (UniProt); Regulation: Type II predominates in tumors (UniProt). Evidence: UniProt P20839, HPA tissue IHC · Approved, HPA ICC-IF · cytosol and rods & rings. Caution: Based on antibodies targeting…, Datasheet A03791-1, PMC9945078, PMC10197894. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IMPDH1/immunohistochemistry-ihc-guide.