This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
INHA / Inhibin alpha chain · IHC design guide
Design Immunohistochemistry for INHA
Plan chromogenic IHC for INHA in paraffin sections using the catalog antibody (datasheet A02413-3). Assess cytoplasmic staining in adrenal and gonadal cells (HPA tissue IHC), accounting for secretion when interpreting the pattern (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
INHA 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 adrenal and gonadal cells (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in adrenal, Sertoli and granulosa cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02413-3) | |
| Positive control | Adrenal gland+2 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 | Secreted protein may differ in location from its RNA (HPA tissue IHC) | |
| Regulation | Adrenal expression extends in hyperplasia (UniProt) | |
| Isoform / epitope | No isoforms annotated; cleavage creates a mature chain (UniProt) |
Section 1
Recommended INHA IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A02413-3) with two published INHA chromogenic IHC protocols (PMC3777642; PMC11155563).
| Sample | Paraffin-embedded rat testis tissue; fixative not specified (datasheet A02413-3) |
| Fixation | Image fixative and duration unreported (datasheet A02413-3); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A02413-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02413-3) |
| Primary antibody | Rabbit anti-INHA, 2-5μg/ml (datasheet A02413-3) |
| Primary incubation | Overnight at 4 °C (datasheet A02413-3) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02413-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | INHA-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Distinct cytoplasmic expression in adrenal gland, Sertoli cells in testis and granulosa cells in ovary. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet A02413-3); the nasopharyngeal carcinoma protocol used 10 mM citrate (PMC11155563).
Section 2
What Is the Expected INHA Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic INHA staining in adrenal glandular cells and testicular and ovarian cells (HPA: Enhanced tissue IHC). The HPA profile names Sertoli cells and granulosa cells, while its scored tissue entries name Leydig cells and follicle cells; assess the identified cell population rather than treating those labels as interchangeable (HPA: tissue IHC). INHA is secreted and has no transmembrane segment, so staining need not outline the cell membrane (UniProt P05111).
| Distinct cytoplasmic staining in adrenal glandular cells, with signal in the expected testicular or ovarian population. | This fits the reported tissue profile (HPA: Enhanced tissue IHC). Use tissue-specific expectations: adrenal glandular cells and Leydig cells are scored High, whereas ovarian follicle cells are scored Medium (HPA: tissue IHC). |
| Predominantly nuclear or sharp membrane-rim staining replaces the expected cytoplasmic pattern. | Treat the compartment mismatch as a possible artefact and review controls before scoring it as INHA: the tissue IHC profile is cytoplasmic, and INHA has no transmembrane segment (HPA: tissue IHC; UniProt P05111 topology). |
| Strong staining appears mainly in a cell population reported as not detected, while expected cells remain unstained. | Consider antibody cross-reactivity or endogenous detection activity (general IHC practice). For example, HPA reports no staining in adipocytes of adipose tissue; its negative entry applies to those cells, not every cell in that tissue (HPA: tissue IHC). |
| Pale, widespread chromogen obscures cell boundaries across the section. | Treat this as background until controls establish a cell-specific signal (general IHC practice). Diffuse staining alone does not reproduce HPA's distinct cytoplasmic profile in adrenal, testicular and ovarian cells (HPA: tissue IHC). |
| No staining appears in an adrenal gland section expected to provide a positive control. | The result is inconclusive until the staining run is checked: HPA scores adrenal glandular cells High, but UniProt limits normal adrenal cortical expression to the zona reticularis and innermost zona fasciculata (HPA: tissue IHC; UniProt P05111 tissue specificity). |
💡Expected INHA appearanceCall the result consistent with INHA when distinct cytoplasmic signal occurs in the expected adrenal, testicular or ovarian cells at a tissue-appropriate level; predominant nuclear, membrane-rim or indiscriminate staining is suspect (HPA: Enhanced tissue IHC; UniProt P05111 topology).
| Adrenal sampling (UniProt P05111 tissue specificity) | Normal cortical expression is limited to the zona reticularis and innermost zona fasciculata; a section missing those regions may be a poor positive control (UniProt P05111). |
| Secreted protein and processing (UniProt P05111) | INHA has a signal peptide and is processed into an inhibin alpha chain; RNA and tissue protein locations can differ for secreted proteins (UniProt P05111 processing; HPA: reliability description). |
| Glycosylation and antibody epitope (UniProt P05111) | Three glycosylation sites are annotated, but the supplied sources do not locate the catalog antibody's epitope or establish an effect on IHC staining (UniProt P05111 glycosylation). |
| Antibody validation (HPA: antibodies) | HPA019141 and CAB000047 have Enhanced IHC status; HPA048834 has no listed IHC status. Validation for one antibody does not establish another antibody's performance (HPA: antibodies). |
| IF/ICC appearance (HPA: subcellular ICC-IF) | What pattern should an IF/ICC image show? HPA reports mainly vesicles, with additional Golgi localization; use the separate IF/ICC guide for that application (HPA: subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| Adrenal positive control is blank. | The sampled area may lack the cortical zones with reported expression (UniProt P05111 tissue specificity). | Confirm that the section contains the zona reticularis or innermost zona fasciculata, then review run controls and detection steps (UniProt P05111; general IHC practice). |
| Signal is weak in ovarian follicle cells compared with adrenal glandular cells. | The reported staining levels differ: Medium in ovarian follicle cells and High in adrenal glandular cells (HPA: tissue IHC). | Judge each tissue against its own reported level and check a positive control before changing staining conditions (HPA: tissue IHC; general IHC practice). |
| The testis result seems inconsistent between cell labels. | HPA's summary names Sertoli cells, while its scored positive entry names Leydig cells (HPA: tissue IHC). | Identify the stained cell population on the section and report it explicitly; do not equate the two HPA labels (HPA: tissue IHC). |
| Nuclear or membrane-rim signal dominates. | The pattern conflicts with the reported cytoplasmic tissue signal and lack of a transmembrane segment (HPA: tissue IHC; UniProt P05111 topology). | Review the negative control and antibody validation, then score only a reproducible, cell-specific pattern (general IHC practice; HPA: antibodies). |
| Widespread chromogen appears in a negative-control section. | Endogenous detection activity or nonspecific reagent binding can create background (general IHC practice). | Check the detection-only control, blocking step and wash conditions before interpreting tissue staining (general IHC practice). |
| A section is called negative because it lacks extracellular staining. | Secretion does not require an extracellular deposit to be visible; HPA describes distinct cytoplasmic tissue staining (UniProt P05111; HPA: tissue IHC). | Assess the expected cells for cytoplasmic signal and compare with an appropriate positive control (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing INHA staining
A02413-3 · IHCIHC analysis of Inhibin alpha/INHA using anti-Inhibin alpha/INHA antibody (A02413-3). Inhibin alpha/INHA was detected in paraffin-embedded s
A02413-3 · IHCIHC analysis of Inhibin alpha/INHA using anti-Inhibin alpha/INHA antibody (A02413-3). Inhibin alpha/INHA was detected in a paraffin-embedded
A02413-3 · IFIF analysis of Inhibin alpha/INHA using anti-Inhibin alpha/INHA antibody (A02413-3). Inhibin alpha/INHA was detected in immunocytochemical s
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000123999-INHA/tissue/cerebral+cortex#imid_74962
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000123999-INHA/tissue/adrenal+gland#imid_74930
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000123999-INHA/tissue/colon#imid_64268
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000123999-INHA/tissue/liver#imid_64261
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA019141; green = INHA, blue = nucleus, red = microtubules.
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA019141; green = INHA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for INHA IHC & IF
🧪Run testis first and look for staining in Leydig cells (HPA: High in testis Leydig cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected); on the testis slide, compare Leydig cells with non-Leydig cells for background staining rather than assuming every other cell type is negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show INHA in U2OS, HEL, K-562, hTERT-RPE1 (serum starved), with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a host-matched rabbit IgG isotype control with clonality matched if known (caption: rabbit primary; clonality unreported); and INHA knockout tissue or peptide competition if the relevant material is available. For chromogenic detection, check endogenous peroxidase and endogenous biotin background, especially with the caption’s biotin-based detection system (caption: biotinylated secondary and streptavidin–biotin complex).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A02413-3 paraffin-section caption does not state a fixative (caption: fixative unreported). That caption uses heat retrieval in EDTA at pH 8.0 for rat testis, but does not establish that retrieval is required (caption: EDTA retrieval); endogenous biotin may complicate its biotin-based chromogenic readout (caption: streptavidin–biotin complex). The supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC, although ICC-IF images show vesicular and additional Golgi localization (HPA: subcellular ICC-IF).
HPA tissue IHC evidence for INHA
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Secreted protein, tissue location of RNA and protein is expected to differ.). 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 → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced INHA IHC Tips
Troubleshoot INHA staining in paraffin sections by checking retrieval, cell identity, compartment, controls and scoring before interpreting signal intensity (datasheet A02413-3; HPA tissue IHC).
How should I retrieve INHA when staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A02413-3). The pictured rat testis section used that retrieval before 2 μg/mL primary antibody overnight at 4°C (caption A02413-3). Keep heating and cooling identical across test and control sections; record the cycle because the caption gives no duration (caption A02413-3; standard IHC practice). If signal remains weak, vary heating duration on adjacent sections while holding antibody and detection conditions constant; check tissue integrity too (standard IHC practice). Judge recovery against the reported cytoplasmic testis pattern, accounting for the distinct Sertoli-cell profile and high Leydig-cell listing (HPA tissue IHC).
Could fixation explain variable INHA staining between paraffin blocks?
The selected paraffin-section caption does not report a fixative, so INHA-specific fixation sensitivity is unknown (caption A02413-3). Document fixation and processing for each block, then run matched control sections alongside the affected sections (standard IHC practice). If staining changes, compare sections using EDTA pH 8.0 retrieval and 2 μg/mL primary under matched detection conditions (datasheet A02413-3; standard IHC practice). Score tissue preservation separately from cytoplasmic signal, since damaged morphology makes cell-level comparison unreliable (standard IHC practice; HPA tissue IHC). Do not attribute a difference to INHA glycosylation or cleavage without a controlled fixation comparison (UniProt P05111 processing; standard IHC practice).
Which compartment and cells should show convincing INHA staining?
Expect cytoplasmic staining in adrenal gland, testicular Sertoli cells and ovarian granulosa cells (HPA tissue IHC). INHA is secreted and has no transmembrane segment, making an exclusively sharp plasma-membrane pattern discordant with its annotation (UniProt P05111 subcellular; UniProt P05111 topology). Vesicles are its main approved location in cell imaging, with additional Golgi localisation (HPA subcellular). Use the counterstain to identify producer cells before scoring; extracellular signal alone cannot establish which cell produced the protein (standard IHC practice; UniProt P05111 subcellular). In testis, record cell identity carefully: the profile names Sertoli cells, while the positive-cell table lists high Leydig-cell staining (HPA tissue IHC).
How do processing and glycosylation affect interpretation of INHA epitopes?
INHA has 0 annotated isoforms, but its precursor undergoes signal-peptide and propeptide processing before the mature chain at residues 233–366 (UniProt P05111 processing; UniProt P05111 isoforms). Glycosylation sites are annotated at residues 146, 268 and 302 (UniProt P05111 glycosylation). The supplied caption identifies the antibody and staining conditions, but gives no epitope, so mature-chain recognition cannot be assumed (caption A02413-3). If compartment or cell pattern is unexpected, compare staining with an independently validated antibody of known epitope and matched tissue controls (standard IHC practice). Report the observed staining as INHA immunoreactivity rather than a particular processed form unless the epitope is established (UniProt P05111 processing; standard IHC practice).
How can I check an INHA pattern by multiplex immunofluorescence?
For a parallel IF experiment, pair INHA with a validated marker for the expected cell type, such as a Sertoli-cell marker in testis (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores after examining unstained tissue autofluorescence, and include single-label controls for channel bleed-through (standard IF practice). Because INHA is secreted and appears in vesicles and Golgi, intracellular access may require permeabilisation; optimise it against the antibody’s actual epitope location (UniProt P05111 subcellular; HPA subcellular; standard IF practice). The catalog antibody’s paraffin IHC caption does not establish its IF performance, so validate its IF staining separately (caption A02413-3; standard IF practice). Compare cellular overlap and compartment pattern with the chromogenic result without treating either image alone as proof of cell identity (standard IF/IHC practice).
What should I check when INHA DAB staining is diffuse?
First compare a no-primary section with the stained section to separate detection-system colour from antibody-dependent signal (standard IHC practice). The pictured method used 10% goat serum, a biotinylated secondary, streptavidin-biotin detection and DAB (caption A02413-3). With this detection scheme, check endogenous biotin and include a peroxidase block when using DAB; these are general chromogenic controls, not INHA-specific findings (standard IHC practice). If background persists, titrate around the reported 2 μg/mL primary condition while keeping retrieval and development matched (caption A02413-3; standard IHC practice). Prefer cell-associated cytoplasmic staining over uniform surface colour when assessing the expected pattern (HPA tissue IHC; standard IHC practice).
How should I quantify INHA staining across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue compartment before measuring signal, since adrenal, testicular and ovarian staining has distinct cellular assignments (HPA tissue IHC). For chromogenic sections, record percentage of positive target cells and an intensity category from 0–3, then calculate an H-score from 0–300 (standard IHC practice). Keep the intensity thresholds, counterstain, imaging settings and region-selection rules consistent across the batch (standard IHC practice). Normalise positive-cell counts to all evaluable cells of the same type, or report positive-cell density per mm² of evaluable tissue (standard IHC practice). Exclude folds and damaged regions, and report replicate fields or sections so sampling variation remains visible (standard IHC practice).
How do I distinguish true INHA staining from artefact?
Seek cell-associated cytoplasmic staining in an expected population, supported by a positive control such as the pictured rat testis section (HPA tissue IHC; caption A02413-3). A membrane-only pattern conflicts with INHA’s secreted, non-transmembrane annotation, while extracellular material alone does not identify a producer cell (UniProt P05111 topology; UniProt P05111 subcellular; standard IHC practice). Treat staining confined to tissue edges, folds or necrotic regions as suspect and inspect the corresponding morphology (standard IHC practice). Check a no-primary section and peroxidase-block control when DAB colour appears broadly or in unexpected cells (standard IHC practice). Interpret negative controls by their annotated cells: appendix glandular cells and adipose adipocytes are listed as not detected (HPA tissue IHC).
Boster reagents
Best INHA / Inhibin alpha chain IHC Antibodies
A02413-3 has IHC images of rat and human paraffin-embedded testis and an IF image of U20S cells (A02413-3 image captions). Its listed reactivity includes human, mouse and rat (catalog).
Real IHC data
Anti-Inhibin alpha/INHA Antibody ®
Cat # A02413-3
A02413-3 will render with a rat testis IHC figure; its additional captions show human testis IHC and U20S cell IF (A02413-3 image captions). M02413-2 will render as a human-reactive IHC antibody, but no IHC or IF image is supplied (M02413-2 catalog).
Which to pick: Choose A02413-3 for tissue IHC supported by rat and human paraffin-section images; the captions do not report a fixative (A02413-3 IHC captions). Choose A02413-3 for IF/ICC and cross-species work because those applications and human, mouse and rat reactivity are listed; its IF image shows U20S cells (A02413-3 catalog and IF caption). M02413-2 is a rabbit monoclonal with listed human reactivity and IHC use, but no tissue image or IF/ICC application is supplied (M02413-2 catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P05111 (INHA_HUMAN, Inhibin alpha chain).
- Human Protein Atlas. INHA tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. INHA subcellular location (ICC-IF): Mainly localized to vesicles. In addition localized to the Golgi apparatus..
- Human Protein Atlas. INHA antibody validation summary (3 antibodies).
- Expression of Wnt and TGF-β pathway components and key adrenal transcription factors in adrenocortical tumors: association to carcinoma aggressiveness. Pathology, research and practice 2013 — PMC3777642.
- Study on the value of Inhibin B in the diagnosis of nasopharyngeal carcinoma and its correlation with traditional Chinese medicine syndromes: An observational study. Medicine 2024 — PMC11155563.
- Testis exposure to unopposed/elevated activin A in utero affects somatic and germ cells and alters steroid levels mimicking phthalate exposure. Frontiers in endocrinology 2023 — PMC10505732.
- PubMed PMID:3016724 — UniProt-cited evidence.
- PubMed PMID:3758355 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for INHA (Inhibin alpha chain). Expected localisation Cytoplasmic staining in adrenal and gonadal cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02413-3). Validated IHC antibody: A02413-3 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat testis tissue; fixative not specified (datasheet A02413-3); 2. Fixation: Image fixative and duration unreported (datasheet A02413-3); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A02413-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02413-3); 8. Primary antibody: Rabbit anti-INHA, 2-5μg/ml (datasheet A02413-3); 9. Primary incubation: Overnight at 4 °C (datasheet A02413-3). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Secreted protein may differ in location from its RNA (HPA tissue IHC); Isoform / epitope: No isoforms annotated; cleavage creates a mature chain (UniProt); Regulation: Adrenal expression extends in hyperplasia (UniProt). Evidence: UniProt P05111, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to vesicles. In addition localized to the Golgi…, Datasheet A02413-3, PMC3777642, PMC11155563. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/INHA/immunohistochemistry-ihc-guide.