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INHA / Inhibin alpha chain · IHC design guide
Design Immunohistochemistry for INHA
Plan paraffin section IHC for INHA using its observed cytoplasmic tissue pattern (HPA tissue IHC). Compare adrenal gland, testis and ovary staining while accounting for secretion and processing (HPA tissue IHC; 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 (HPA tissue IHC); secreted protein (UniProt) | |
| Staining pattern | Cytoplasm in adrenal, testis Sertoli and ovarian 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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Secretion can separate tissue protein from its RNA source (HPA tissue IHC) | |
| Regulation | Adrenal expression extends in hyperplasia (UniProt) | |
| Isoform / epitope | No isoforms annotated; cleavage yields a mature chain at 233–366 (UniProt) |
Section 1
Recommended INHA IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet A02413-3). Two published INHA protocols provide paraffin-section examples from adrenocortical tumors and nasopharyngeal tissues (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 retrieval for the catalog antibody (datasheet A02413-3). A published nasopharyngeal tissue protocol used 10 mM citrate (PMC11155563).
Section 2
What Is the Expected INHA Staining Pattern?
INHA is a secreted protein with no transmembrane segment (UniProt P05111), and HPA reports distinct cytoplasmic IHC staining in adrenal gland, testis and ovary (HPA: Enhanced tissue reliability). Assess the signal in the annotated cells: adrenal glandular cells and testis Leydig cells stain strongly, while ovary follicle cells show medium staining (HPA: tissue IHC). HPA’s profile also describes Sertoli and granulosa cell expression (HPA: tissue IHC profile).
| Strong cytoplasmic staining in adrenal glandular or testis Leydig cells; medium staining in ovary follicle cells (HPA: tissue IHC). | This matches HPA’s cell-level IHC results. Score intensity within the relevant cells and compare it with adjacent tissue; do not treat every cell in a positive organ as an expected positive (HPA: tissue IHC; general IHC practice). |
| Predominantly nuclear or sharply membranous staining, with little cytoplasmic signal. | That distribution conflicts with HPA’s cytoplasmic tissue pattern and UniProt’s secreted, non-transmembrane annotation. Treat it as suspect until controls and an independently validated antibody support it (HPA: tissue IHC; UniProt P05111 topology; general IHC practice). |
| Strong staining in adipocytes or appendix glandular cells. | HPA reports INHA as not detected in those specified cells. Check whether the signal follows cell boundaries or appears in controls before interpreting it as INHA; unexpected staining can reflect cross-reactivity or detection background (HPA: tissue IHC; general IHC practice). |
| Diffuse color across cells and stroma, without a recognizable cellular pattern. | The slide cannot establish the HPA-described cytoplasmic pattern. Review the no-primary control, blocking, washing and chromogen development for nonspecific or endogenous signal before assigning expression (HPA: tissue IHC; general IHC practice). |
| No staining in adrenal glandular cells or testis Leydig cells. | Both are high-staining reference cell populations in HPA’s tissue IHC data. First check that the intended cells are present; then review the antibody, retrieval and detection workflow. An unstained positive control makes a negative test section inconclusive (HPA: tissue IHC; general IHC practice). |
💡Expected INHA appearanceCall positive when cytoplasmic signal is strong in adrenal glandular or testis Leydig cells, or medium in ovary follicle cells; isolated nuclear, membranous or diffuse stromal color is suspect (HPA: tissue IHC; UniProt P05111 topology).
| Cell-level reference pattern | HPA lists high staining in adrenal glandular and testis Leydig cells, and medium staining in ovary follicle cells. Its narrative also names Sertoli and granulosa cells; preserve that distinction when selecting and scoring fields (HPA: tissue IHC). |
| Secretory location | INHA has a signal peptide, is secreted and has no annotated transmembrane segment (UniProt P05111). A cytoplasmic production-site pattern is consistent with HPA tissue IHC; staining alone cannot establish where secreted protein travels (HPA: tissue IHC; UniProt P05111). |
| Protein processing and epitope | UniProt maps the signal peptide to residues 1–18, propeptide regions to 19–232 and the inhibin alpha chain to 233–366. If the antibody epitope is known, compare its location with these regions; the supplied record gives no epitope (UniProt P05111). |
| Glycosylation | Three glycosylation sites are annotated at residues 146, 268 and 302 (UniProt P05111). The supplied evidence does not show whether glycosylation changes staining by the catalog antibody; use the sites only when interpreting a documented epitope (UniProt P05111). |
| Antibody validation | HPA019141 and CAB000047 have Enhanced IHC status; HPA048834 has no listed IHC status (HPA: antibodies). Enhanced supports agreement with independent antibodies or orthogonal data, but each new staining run still needs appropriate controls (HPA: antibody-validation summary; general IHC practice). |
| Retrieval and chromogenic detection | Choose antigen retrieval and detection settings from the IHC-validated antibody’s documented protocol, then verify them with positive and no-primary controls (general IHC practice). No INHA-specific fixation sensitivity or retrieval effect is supplied by HPA or UniProt. |
| IF/ICC Q&A | What location can IF/ICC show? HPA reports mainly vesicles, with additional Golgi localization, in its approved ICC-IF data (HPA: subcellular ICC-IF). This informs localization checks; it is not an IHC-P protocol or evidence that every tissue section resolves those structures. |
| Situation | Likely cause | Next action |
|---|---|---|
| The adrenal positive control is blank. | The expected glandular-cell signal is high (HPA: tissue IHC); an absent control signal leaves a workflow or reagent problem unresolved (general IHC practice). | Confirm glandular cells are present, then check antibody identity, documented retrieval conditions, detection reagents and run controls before scoring test sections negative (HPA: tissue IHC; general IHC practice). |
| Only nuclei or cell outlines stain. | That pattern differs from HPA’s cytoplasmic IHC description and the secreted, non-transmembrane annotation (HPA: tissue IHC; UniProt P05111 topology). | Compare the no-primary control and an expected positive tissue; review antibody specificity and chromogen localization before calling the signal INHA (general IHC practice). |
| Adipocytes or appendix glandular cells appear strongly positive. | Those cell populations are listed as not detected, so unexpected color needs verification; the image alone cannot distinguish cross-reactivity from detection background (HPA: tissue IHC; general IHC practice). | Recheck cell identification, examine the no-primary control and compare a positive reference section under the same detection conditions (general IHC practice). |
| Brown color is widespread without cell-specific contrast. | Diffuse background can obscure the HPA-described cytoplasmic pattern; nonspecific binding or endogenous detection activity are possible general IHC causes (HPA: tissue IHC; general IHC practice). | Inspect no-primary and detection controls; review blocking, washing and chromogen development, then repeat scoring only when the cellular pattern is readable (general IHC practice). |
| Ovary appears weaker than adrenal gland or testis. | HPA grades ovary follicle cells medium, versus high in adrenal glandular and testis Leydig cells; lower intensity can therefore fit the reference pattern (HPA: tissue IHC). | Score the follicle cells themselves and compare slides stained in the same run; use the tissue-specific HPA levels rather than one intensity threshold for all three organs (HPA: tissue IHC; general IHC practice). |
| Testis staining is assigned to different cell types by different reviewers. | HPA’s cell-level table lists Leydig cells as high, while its profile describes Sertoli-cell cytoplasmic expression; the supplied HPA descriptions differ in emphasis (HPA: tissue IHC). | Record the stained cell type and morphology explicitly, review the matching HPA image and antibody validation, and avoid collapsing Leydig and Sertoli observations into one claim (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 adrenal gland first: glandular cells should stain (HPA: High in adrenal gland glandular cells). Run appendix glandular cells as the negative tissue (HPA: Not detected in appendix glandular cells); on the adrenal slide, use adjacent nonglandular cells as internal background comparators and check that they show little or no signal.
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 and a concentration-matched rabbit isotype control for the rabbit catalog antibody (caption: rabbit anti-INHA); where available, use INHA-knockout material or validated peptide competition as a biological specificity control (standard IHC practice). For adrenal DAB staining, quench endogenous peroxidase and check for endogenous biotin background with the biotin-based detection system (caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A02413-3 tissue-IHC caption does not state a fixative (caption: fixative unreported). Paraffin-section IHC with heat retrieval in EDTA at pH 8.0 is documented, but retrieval dependence has not been established; the supplied evidence does not show that frozen sections or IF are easier (caption: EDTA retrieval in paraffin sections). In adrenal sections, assess blood-associated peroxidase background separately from glandular staining (standard IHC practice; HPA: High in adrenal gland glandular cells).
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 by checking retrieval, compartment, cell type and detection controls before comparing signal across sections.
Which retrieval condition should I start with for paraffin section INHA staining?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A02413-3). That condition accompanied detection in a paraffin embedded rat testis section with the catalog antibody at 2 µg/mL (caption A02413-3). Keep heating and cooling consistent across sections, then compare a fresh positive control with the problem section before changing antibody concentration. If staining remains weak, adjust retrieval duration in small steps and inspect tissue integrity; any alternative buffer requires its own validation. The caption does not state the fixative, so this result does not establish performance under a particular fixation condition (caption A02413-3).
Could fixation explain weak or uneven INHA staining?
The available product caption identifies a paraffin embedded section but does not report its fixative (caption A02413-3). Target specific fixation sensitivity is therefore unknown; record the fixative, fixation duration, tissue thickness and processing history for every comparison. Evaluate a known positive section processed alongside the problem section, holding EDTA retrieval at pH 8.0 and antibody concentration at 2 µg/mL initially (caption A02413-3). If both sections stain poorly, check processing and reagent performance before attributing the result to INHA biology. Compare differently fixed samples only after aligning retrieval, detection and scoring conditions, because processing differences can change apparent staining.
Where should convincing INHA signal appear within positive cells?
Expect predominantly cytoplasmic staining in adrenal gland, testicular Sertoli cells and ovarian granulosa cells (HPA tissue IHC profile). A vesicular pattern, with additional Golgi localisation, is supported by cellular imaging (HPA subcellular). INHA has a signal peptide at residues 1–18, lacks a transmembrane segment and is secreted (UniProt P05111); extracellular signal may therefore occur, but it needs careful interpretation. Check whether staining follows plausible cell boundaries and recurs in the expected cell population across intact fields. Treat isolated nuclear staining or uniform staining of every compartment as suspicious until controls and tissue morphology support it.
How can processing and glycosylation affect an INHA epitope in sections?
INHA is synthesised as a 366 amino acid precursor with a signal peptide, propeptide regions and a mature alpha chain spanning residues 233–366 (UniProt P05111). Glycosylation sites are annotated at residues 146, 268 and 302, and no isoforms are annotated (UniProt P05111). Check the antibody's documented immunogen or epitope before assuming that staining reports the mature chain rather than a precursor region. Differences in processing or epitope accessibility could affect recognition, but the supplied evidence does not map the catalog antibody's epitope. Compare staining with an independently characterised antibody only after checking that its epitope and tissue controls address the same question.
How should I adapt the INHA localisation check to multiplex IF?
Pair INHA with a validated marker for the cell population being examined, such as a Sertoli cell marker in testis or a granulosa cell marker in ovary (HPA tissue IHC profile). Choose spectrally separated fluorophores and place the weaker signal in a channel with low tissue autofluorescence; acquire single stained and unstained controls to assess bleed through and autofluorescence. INHA is secreted and lacks a transmembrane segment (UniProt P05111), while cellular images place it mainly in vesicles and also in Golgi (HPA subcellular). Optimise mild permeabilisation for intracellular epitopes, then compare with an unpermeabilised control if extracellular signal is of interest. Validate IF conditions independently of the paraffin section caption.
What should I check when DAB appears diffusely brown?
The documented paraffin section workflow used 10% goat serum, a biotinylated secondary antibody, a streptavidin biotin complex and DAB (caption A02413-3). Run a no primary control and examine whether brown signal remains in the same structures; include appropriate controls for endogenous peroxidase and endogenous biotin when using this detection system. Inspect section edges, folds and damaged areas before calling diffuse color positive. Titrate the primary antibody around the documented 2 µg/mL condition while keeping development time consistent (caption A02413-3). Because INHA is secreted (UniProt P05111), extracellular color alone should be judged against cell associated staining and control sections.
How should I score INHA staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical compartment and eligible cells before scoring: adrenal glandular cells, testicular cells or ovarian follicle cells are supported comparison populations (HPA tissue IHC). Within that region, record the percentage of positive cells and staining intensity, then calculate an H-score using one prespecified intensity scale. Normalise counts to the number of eligible intact cells, or report positive cell density per mm² of viable tissue when cell counting is impractical. Keep retrieval, antibody concentration, chromogen development and image exposure consistent between samples. Score extracellular deposits separately from cell associated cytoplasmic staining because INHA is secreted (UniProt P05111).
How can I distinguish specific INHA staining from artefact?
Look for reproducible cytoplasmic signal in a biologically plausible cell population: adrenal glandular cells are high, while ovarian follicle cells are medium in the supplied tissue profile (HPA tissue IHC). Cellular imaging supports vesicular and additional Golgi localisation (HPA subcellular). Diffuse staining at section edges, necrotic areas or folds deserves comparison with intact tissue and a no primary control. Persistent brown signal in that control suggests detection background, including endogenous enzyme activity in a chromogenic workflow. Interpret extracellular deposits cautiously because INHA is secreted (UniProt P05111), and avoid treating isolated nuclear color as evidence of the expected localisation.
Boster reagents
Best INHA / Inhibin alpha chain IHC Antibodies
The IHC-validated catalog antibody covers human, mouse and rat (catalog: A02413-3 reactivity); its image captions show paraffin-section IHC in rat and human testis and IF/ICC in U20S cells (A02413-3 captions).
Real IHC data
Anti-Inhibin alpha/INHA Antibody ®
Cat # A02413-3
A02413-3 will render with its rat testis paraffin-section IHC figure; its other captions document human testis paraffin-section IHC and U20S-cell IF/ICC (A02413-3 image captions). M02413-2 will render as a human-reactive IHC antibody, with no IHC figure supplied (M02413-2 catalog).
Which to pick: Choose A02413-3 for tissue IHC when a demonstrated paraffin-section procedure matters: its captions document rat and human testis staining, but do not report the fixative (A02413-3 IHC image captions). Choose A02413-3 for IF/ICC or work spanning human, mouse and rat because those applications and species are listed, and its IF caption shows U20S cells; the shown IHC tissue data cover rat and human only (A02413-3 catalog and image captions). M02413-2 offers a rabbit monoclonal for human IHC, though no IHC image or tissue-processing details are 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 (HPA tissue IHC); secreted protein (UniProt). 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 paraffin sections. (standard IHC practice; not target-specific); Caveat: Secretion can separate tissue protein from its RNA source (HPA tissue IHC); Isoform / epitope: No isoforms annotated; cleavage yields a mature chain at 233–366 (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.