This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
GUCY1B1 / Guanylate cyclase soluble subunit beta-1 · IHC design guide
Design Immunohistochemistry for GUCY1B1
This GUCY1B1 IHC-P guide identifies cytoplasmic staining and high-staining neuronal and glandular cells for tissue controls (HPA tissue IHC). Start M32396 at 1:50 (datasheet M32396), then compare staining within the relevant cell types.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GUCY1B1 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 staining in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M32396) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Cervix+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 | Some squamous epithelial cells are unstained (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage unreported (UniProt) |
Section 1
Recommended GUCY1B1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet M32396) is accompanied by one published paraffin-section GUCY1B1 protocol (PMC12229189).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet M32396) |
| Fixation | Image fixative and duration unreported (datasheet M32396); 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 M32396); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M32396) |
| Primary antibody | Rabbit monoclonal (clone 20G57) anti-GUCY1B1, 1:50 (datasheet M32396) |
| Primary incubation | Overnight at 4 °C (datasheet M32396) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M32396) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GUCY1B1-positive staining in neuronal cells of caudate (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet M32396); the published article does not specify retrieval (PMC12229189).
Section 2
What Is the Expected GUCY1B1 Staining Pattern?
GUCY1B1 is a cytoplasmic protein with no transmembrane segment (UniProt Q02153). In paraffin-section IHC, expect cytoplasmic staining in neuronal cells of the caudate and cerebral cortex, cells of the cerebellar molecular layer, and glandular cells of the gallbladder and parathyroid gland (HPA: High in these cell populations). HPA rates the tissue IHC pattern Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC).
| Cytoplasmic chromogen in caudate or cerebral cortex neuronal cells, or cerebellar molecular-layer cells. | This matches the reported High tissue IHC pattern and cytoplasmic profile (HPA: tissue IHC). Judge the named cells against nearby structures; staining somewhere else on the section does not establish the expected cell-specific pattern (general IHC practice). |
| Predominantly nuclear or sharply membrane-restricted staining, with little cytoplasmic signal. | That compartment conflicts with cytoplasmic tissue staining (HPA: tissue IHC), approved cytosolic localization (HPA: ICC-IF), and the absence of a transmembrane segment (UniProt Q02153). Treat it as a possible staining artefact and check morphology and controls before scoring it as GUCY1B1 (general IHC practice). |
| Strong staining in cells reported as undetected, such as lymph-node germinal-center cells. | HPA reports GUCY1B1 as Not detected in those cells (HPA: tissue IHC). Unexpected chromogen may reflect cross-reactivity or endogenous detection activity (general IHC practice); the HPA result is a comparison point, not proof that every specimen must be negative. |
| Diffuse chromogen across cells and extracellular spaces, obscuring cell boundaries. | An uninterpretable wash of color does not reproduce the reported cellular cytoplasmic pattern (HPA: tissue IHC). Excess detection background or nonspecific binding are possible explanations (general IHC practice); assess the no-primary control before assigning any positive cells. |
| No discernible signal in caudate neuronal cells or gallbladder glandular cells. | Both are reported High in tissue IHC (HPA: tissue IHC), so absence warrants an assay check. It does not alone show that the specimen lacks GUCY1B1: tissue handling, antibody performance, and detection can also affect an IHC result (general IHC practice). |
💡Expected GUCY1B1 appearanceCall a result positive when identifiable neuronal, molecular-layer, or glandular cells show cytoplasmic chromogen, potentially High in the HPA-listed positive tissues (HPA: tissue IHC); predominantly nuclear, membrane-restricted, or cell-independent color is suspect (UniProt Q02153; HPA: tissue IHC; general IHC practice).
| Cell and tissue selection | HPA reports High staining in caudate and cortical neuronal cells, cerebellar molecular-layer cells, and gallbladder and parathyroid glandular cells, but Not detected in several other named cell populations (HPA: tissue IHC). Compare the same cell type when evaluating controls. |
| Antibody evidence | The tissue profile is Approved with medium staining–RNA consistency (HPA: tissue IHC). HPA020870 and CAB010890 each have Approved IHC status (HPA: antibodies). These ratings support comparison with the reported pattern but do not establish specificity in every preparation. |
| Compartment and protein form | UniProt places GUCY1B1 in the cytoplasm, reports no transmembrane segment, and lists a chain spanning residues 1–619 with no signal peptide or propeptide (UniProt Q02153). Those annotations support cytoplasmic scoring; they do not identify the catalog antibody's epitope. |
| Isoform coverage | UniProt lists 3 isoforms (UniProt Q02153). The supplied records do not specify which isoforms an IHC antibody recognizes, so a difference between samples cannot be assigned to isoform expression from this staining pattern alone. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in an expected positive cell population. | The result differs from HPA's High staining in caudate neuronal or gallbladder glandular cells (HPA: tissue IHC); the technical cause is undetermined. | Confirm the cell type and slide controls, then review the IHC-validated antibody's supplied dilution and antigen-retrieval directions and the detection run (general IHC practice). No GUCY1B1-specific retrieval condition is supplied here. |
| Broad, hazy chromogen masks the cells. | Nonspecific binding or excessive detection background may obscure the cytoplasmic pattern (general IHC practice; HPA: tissue IHC pattern). | Compare a no-primary control, then review blocking, antibody dilution, washing, and chromogen development under the established IHC workflow (general IHC practice). |
| Predominantly nuclear or membrane-like stain. | The location disagrees with cytoplasmic and cytosolic annotations (UniProt Q02153; HPA: tissue IHC and ICC-IF). | Recheck cellular morphology and counterstain, inspect controls, and repeat with an IHC-approved antibody if the compartment remains discordant (HPA: antibodies; general IHC practice). |
| Strong color in an HPA-undetected cell population. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA reports germinal-center cells as Not detected (HPA: tissue IHC). | Compare the named cells with a positive tissue, inspect a no-primary control, and assess whether the signal follows cell boundaries before interpreting it (general IHC practice). |
| Uneven staining within a section. | This may reflect section or detection variability (general IHC practice); HPA's cell-specific levels do not establish the cause of patchiness in a particular specimen. | Check tissue integrity and whether positive and negative regions track recognizable cell types, then review coverage during reagent application (general IHC practice). |
| Q: Can ICC-IF help assess the expected compartment? | HPA reports approved cytosolic localization in ICC-IF (HPA: subcellular) and Supported ICC status for HPA077706 and CAB010890 (HPA: antibodies). | A: Use that evidence as a localization cross-check; follow the separate IF/ICC guide for assay design. An ICC-IF image does not replace cell-specific interpretation of paraffin-section IHC (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing GUCY1B1 staining
M32396 · IHCIHC analysis of GUCY1B1 using anti-GUCY1B1 antibody (M32396). GUCY1B1 was detected in a paraffin-embedded section of human colon cancer tiss
M32396 · IHCIHC analysis of GUCY1B1 using anti-GUCY1B1 antibody (M32396). GUCY1B1 was detected in a paraffin-embedded section of human colon tissue. Hea
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000061918-GUCY1B1/tissue/cerebral+cortex#imid_5677252
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000061918-GUCY1B1/tissue/colon#imid_5677170
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000061918-GUCY1B1/tissue/liver#imid_5677163
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000061918-GUCY1B1/tissue/pancreas#imid_5677130
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB010890; green = GUCY1B1, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB010890; green = GUCY1B1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GUCY1B1 IHC & IF
🧪Run caudate first and score neuronal cells, which HPA rates High for GUCY1B1 (HPA: Caudate neuronal cells, High). Use cervix squamous epithelium as a negative tissue (HPA: Cervix squamous epithelial cells, Not detected); on the caudate slide, cells without neuronal staining should remain at background rather than showing diffuse chromogen.
Positive control tissue: Caudate (Neuronal cells, HPA High)
Negative control tissue: Cervix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GUCY1B1 in U-251MG, U2OS, HEL, JURKAT, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a rabbit IgG isotype control matched to the primary antibody’s clonality, which is unreported (selected SKU M32396 caption: rabbit primary); use GUCY1B1 knockout material as a biological negative where available. Quench endogenous peroxidase for chromogenic IHC and assess tissue autofluorescence separately if using IF (standard IHC/IF practice).
⚠️Feasibility: The selected paraffin-section caption reports heat retrieval in EDTA at pH 8.0 and a 1:50 primary dilution, but does not establish that retrieval is required or report the fixative (selected SKU M32396 caption). No target-specific fixation window or fixation effect is reported in the supplied evidence. Paraffin IHC has a documented example (selected SKU M32396 caption); whether frozen sections or IF are easier is unreported, and caudate autofluorescence should be checked if IF is used (standard IF practice).
HPA tissue IHC evidence for GUCY1B1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GUCY1B1 IHC Tips
Troubleshoot chromogenic IHC for GUCY1B1 in paraffin sections using the catalog image conditions and independent tissue and localisation evidence.
Which retrieval condition should I start with for GUCY1B1 paraffin IHC?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M32396). The catalog image used that condition before staining human colon cancer tissue, but its caption does not report a fixative or heating duration (datasheet M32396). If staining is weak, check that sections remained covered by retrieval buffer and compare heated with unheated sections while holding antibody concentration and detection constant (standard IHC practice). Use a positive reference section in each run, since HPA reports high staining in caudate neuronal cells and cerebellar molecular layer cells (HPA: High in these cell populations).
How should I assess whether fixation is masking GUCY1B1?
Target specific fixation sensitivity is unknown: the M32396 tissue caption describes a paraffin section but does not identify its fixative (datasheet M32396). Record the actual fixative, fixation duration, and processing history for each specimen before attributing weak staining to antigen loss (standard IHC practice). Compare sections with documented processing under the same pH 8.0 EDTA retrieval and detection conditions, then look for a consistent change across replicates (datasheet M32396; standard IHC practice). Include a section with an established cytoplasmic staining pattern as a run control, while recognising that processing differences can confound tissue comparisons (HPA: cytoplasmic expression in most tissues; standard IHC practice).
Where should convincing GUCY1B1 staining appear in a tissue section?
Expect predominantly cytoplasmic staining: UniProt assigns GUCY1B1 to the cytoplasm, and HPA reports cytoplasmic expression in most tissues (UniProt Q02153: subcellular location; HPA: tissue profile). The protein has no transmembrane segment, while HPA's subcellular assessment places it in the cytosol (UniProt Q02153: topology; HPA: Cytosol approved). Check whether signal fills cell bodies rather than tracing plasma membranes or appearing only in nuclei, and compare its distribution with the tissue architecture (standard IHC practice). HPA reports high staining in caudate and cerebral cortex neuronal cells, providing cell type references for evaluating localisation (HPA: High in neuronal cells).
Can this antibody distinguish GUCY1B1 isoforms or epitope loss?
Treat isoform discrimination as unresolved unless the antibody's mapped epitope and isoform reactivity are documented; the supplied M32396 caption does not provide either (datasheet M32396). UniProt lists 3 isoforms and places the H-NOX, PAS H-NOXA type, and guanylate cyclase domains at residues 1–184, 204–338, and 421–554, respectively (UniProt Q02153: isoforms and domains). Check whether any available epitope sequence is shared across the isoforms before interpreting a negative section as absence of all GUCY1B1 forms (standard IHC practice). Compare staining with an independently validated antibody to a different epitope when isoform coverage or epitope masking materially affects the conclusion (standard IHC practice).
How should I adapt GUCY1B1 localisation checks to multiplex IF?
Treat IF as a separately optimised application: the M32396 caption documents chromogenic staining of a paraffin section, while HPA lists ICC/IF images without establishing M32396 IF performance (datasheet M32396; HPA: subcellular record). Multiplex with a marker identifying the expected cell population, such as neuronal cells when evaluating caudate, and check whether GUCY1B1 signal occupies their cytoplasm (HPA: High in caudate neuronal cells; UniProt Q02153: cytoplasm). Choose fluorophores after inspecting unstained tissue for autofluorescence, and include single stain controls for channel bleed through (standard IF practice). Optimise permeabilisation for a cytosolic epitope because GUCY1B1 has no transmembrane segment (HPA: Cytosol approved; UniProt Q02153: topology).
How can I distinguish diffuse GUCY1B1 staining from chromogenic background?
Run a no primary control and examine whether DAB appears in the same structures as the test section; diffuse control staining points to detection background (standard IHC practice). The catalog image used 10% goat serum blocking, 1:50 primary antibody overnight at 4°C, and peroxidase based DAB detection (datasheet M32396). Check the peroxidase block as a general chromogenic IHC step, particularly where endogenous enzyme activity could produce colour without specific antibody binding (standard IHC practice). Compare signal in intact cell cytoplasm with precipitate, tissue edges, and damaged areas before changing the antibody concentration (HPA: cytoplasmic expression; standard IHC practice).
What should I score when comparing GUCY1B1 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue compartment before scoring, then quantify cytoplasmic staining rather than whole section colour (HPA: cytoplasmic expression in most tissues; standard IHC practice). An H-score can combine the percentage of cells at each intensity, while percentage positive cells or positive cell density per mm² can suit a specific question (standard IHC practice). Normalise counts to evaluable cells or viable tissue area, and keep section thickness, retrieval, imaging, and threshold settings consistent across groups (standard IHC practice). Report cell types separately where relevant because HPA records high neuronal staining in caudate but no detected staining in lymph node germinal center cells (HPA: tissue IHC).
What evidence supports a true GUCY1B1 positive IHC result?
A credible positive shows cytoplasmic staining in identifiable intact cells and is absent from the matched no primary control (UniProt Q02153: cytoplasm; standard IHC practice). High signal in caudate neuronal cells or cerebellar molecular layer cells fits HPA observations, whereas staining confined to lymph node germinal center cells warrants closer validation (HPA: tissue IHC). Question predominantly nuclear or membrane rim staining, sharp edge gradients, necrotic debris, and DAB signal reproduced by the no primary control (HPA: Cytosol approved; standard IHC practice). Interpret staining as protein detection, not direct proof of active NO to cGMP signalling, because GUCY1B1 functions within an alpha beta heterodimer (UniProt Q02153: function and subunit).
Boster reagents
Best GUCY1B1 / Guanylate cyclase soluble subunit beta-1 IHC Antibodies
The catalog includes IHC images of human colon, colon cancer, ovarian cancer and mouse brain sections (M32396 image captions); IF/ICC is listed for human, mouse and rat (A32396-1 applications/reactivity).
Real IHC data
Anti-Guanylyl Cyclase beta 1 Rabbit Monoclonal Antibody
Cat # M32396
M32396 is the SKU with a rendered card and its own IHC figure, showing a paraffin section of human colon cancer (card payload: M32396). Its additional IHC captions cover human colon, human ovarian cancer and mouse brain paraffin sections; rat reactivity is listed without a supplied IHC image (catalog: M32396 image captions/reactivity).
Which to pick: Choose monoclonal M32396 for tissue IHC when an illustrated paraffin-section procedure is useful (catalog: M32396 clone 20G57; M32396 IHC image captions). Choose polyclonal A32396-1 for IF/ICC because those applications are listed, although no IF image caption is supplied (catalog: A32396-1 applications/clone/IF image alts). Both list human, mouse and rat reactivity; M32396 has illustrated IHC in human and mouse, and its paraffin-section captions do not report the fixative (catalog: M32396 and A32396-1 reactivity; M32396 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q02153 (GCYB1_HUMAN, Guanylate cyclase soluble subunit beta-1).
- Human Protein Atlas. GUCY1B1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. GUCY1B1 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. GUCY1B1 antibody validation summary (3 antibodies).
- Smooth Muscle Cell-Derived Fibronectin Promotes an Atheroprotective Smooth Muscle Cell Phenotype Associated With Altered NO-cGMP Signaling. Journal of the American Heart Association 2025 — PMC12229189.
- PubMed PMID:1352257 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GUCY1B1 (Guanylate cyclase soluble subunit beta-1). Expected localisation Cytoplasmic staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M32396). Validated IHC antibody: M32396 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Neuronal cells); negative Cervix. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet M32396); 2. Fixation: Image fixative and duration unreported (datasheet M32396); 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 M32396); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M32396); 8. Primary antibody: Rabbit monoclonal (clone 20G57) anti-GUCY1B1, 1:50 (datasheet M32396); 9. Primary incubation: Overnight at 4 °C (datasheet M32396). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Some squamous epithelial cells are unstained (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage unreported (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q02153, HPA tissue IHC · Approved, HPA ICC-IF · cytosol., Datasheet M32396, PMC12229189. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GUCY1B1/immunohistochemistry-ihc-guide.