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POMC / Pro-opiomelanocortin · IHC design guide
Design Immunohistochemistry for POMC
Plan paraffin IHC for POMC using anterior pituitary cells as a positive reference (HPA tissue IHC). The guide covers staining controls and interpretation of a secreted, processed precursor (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
POMC 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 | Secreted (UniProt); anterior pituitary cells stain (HPA tissue IHC) | |
| Staining pattern | Anterior pituitary cells stain; compartment unreported (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet PB9039) | |
| Positive control | Pituitary gland | |
| 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 stain away from its RNA source (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | No isoforms annotated; processing may affect epitope detection (UniProt) |
Section 1
Recommended POMC IHC & IF Protocols
The catalog antibody has a paraffin-section IHC protocol (datasheet PB9039). Published POMC staining conditions below cover mouse brain, rat brainstem, and sheep tissue sections (PMC8228876; PMC3583363; PMC13217381).
| Sample | Paraffin-embedded mouse kidney tissues; fixative not specified (datasheet PB9039) |
| Fixation | Image fixative and duration unreported (datasheet PB9039); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet PB9039) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9039) |
| Primary antibody | Rabbit anti-POMC, 0.5-1μg/ml (datasheet PB9039) |
| Primary incubation | Overnight at 4 °C (datasheet PB9039) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9039) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | POMC-positive staining in cells in anterior of pituitary gland (HPA tissue IHC: High). HPA tissue profile: Highly expressed in the anterior lobe of the pituitary gland. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6 heat retrieval for paraffin sections (datasheet PB9039). Treat published primary dilutions as starting points for their stated tissue preparations (PMC8228876; PMC3583363; PMC13217381).
Section 2
What Is the Expected POMC Staining Pattern?
POMC is a secreted hormone precursor with no transmembrane segment (UniProt P01189 topology). In paraffin section IHC, expect the clearest cellular staining in anterior pituitary cells, where HPA reports high expression (HPA: High in anterior pituitary cells). HPA rates the tissue pattern Enhanced, while noting that a secreted protein’s RNA and protein locations may differ (HPA: Enhanced reliability; secreted protein caveat).
| Strong cytoplasmic staining in a population of anterior pituitary cells. | This fits the reported High anterior pituitary signal (HPA: High in anterior pituitary cells). A cytoplasmic or granular appearance is compatible with a secreted precursor; the supplied IHC record does not define a required granule pattern (UniProt P01189: secreted; HPA: tissue IHC). |
| Predominantly crisp nuclear or cell-surface staining, with little cytoplasmic signal. | Treat this as discordant with the expected cellular pattern: POMC is secreted and has no transmembrane segment (UniProt P01189 topology). Review antibody specificity and detection controls before scoring these compartments as POMC positive (general IHC practice). |
| Strong staining of adipocytes or adrenal glandular cells. | HPA reports POMC as Not detected in those sampled cell types (HPA: adipose tissue adipocytes; adrenal gland glandular cells). Check for cross-reactivity or endogenous detection activity (general IHC practice). HPA negatives describe observed staining, not proof that every cell is biologically incapable of expressing POMC. |
| Diffuse haze across cells, stroma, or much of the section. | A widespread haze cannot by itself establish which cells contain POMC (general IHC practice). Because POMC is secreted, extracellular signal also needs cautious interpretation; it does not identify its cell of origin (UniProt P01189: secreted). Compare a matched detection control and tissue morphology. |
| No convincing stain in anterior pituitary cells. | This conflicts with HPA’s High anterior pituitary result (HPA: High in anterior pituitary cells). First assess tissue identity and whether the detection system worked (general IHC practice). An absent signal alone does not distinguish a technical failure from an antibody epitope mismatch. |
💡Expected POMC appearanceCall positive when anterior pituitary cells show convincing strong cellular staining consistent with a secreted precursor (HPA: High in anterior pituitary cells; UniProt P01189: secreted); isolated nuclear or sharp surface staining warrants specificity review (UniProt P01189 topology).
| Tissue choice | Anterior pituitary provides the documented high signal, while the listed adipocytes and adrenal glandular cells are reported as not detected (HPA: tissue IHC). Use those observations to interpret the slide; they do not establish a universal negative tissue. |
| Secretion and cell of origin | POMC is secreted, and HPA warns that protein location can differ from RNA location (UniProt P01189: secreted; HPA: reliability description). Extracellular staining alone therefore cannot identify the cell that synthesized it. |
| Precursor processing and epitope | POMC is a hormone precursor with a signal peptide and annotated modifications (UniProt P01189: function, processing, modified residues). The supplied record gives no catalog antibody epitope, so it cannot establish which precursor or processed products the stain detects. |
| Antibody evidence | HPA lists 4 antibodies with Enhanced IHC status: HPA046135, HPA063644, CAB002762, and CAB079881 (HPA: antibody IHC validation). This supports the reported tissue pattern, but does not validate an unlisted antibody or a particular staining protocol. |
| IF/ICC? | The same secreted-protein localization caveat applies when interpreting IF/ICC (UniProt P01189: secreted). HPA supplies no main subcellular location, ICC-IF images, or ICC validation for the listed antibodies (HPA: subcellular record; antibody ICC fields). Assess IF/ICC on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| Anterior pituitary section has no signal. | The result conflicts with the reported high staining, but its cause is undetermined (HPA: High in anterior pituitary cells). | Verify tissue identity, section integrity, primary antibody use, and detection controls (general IHC practice). Review antigen retrieval against the antibody’s IHC-P instructions; no POMC-specific retrieval effect is established here. |
| Negative-control section develops chromogen. | Detection chemistry or endogenous activity may contribute signal (general IHC practice). | Review the matched control, endogenous-activity blocking, and detection reagents before interpreting cellular staining (general IHC practice). HPA’s POMC tissue pattern does not identify the source of this background. |
| Adipocytes or adrenal glandular cells stain strongly. | That differs from HPA’s not-detected observations in these cell types (HPA: adipose tissue; adrenal gland). | Compare morphology and matched controls, then assess antibody specificity and endogenous detection activity (general IHC practice). Do not call the staining POMC solely because chromogen is present. |
| Signal is mainly nuclear or sharply outlines cell membranes. | That distribution is difficult to reconcile with a secreted protein lacking a transmembrane segment (UniProt P01189 topology). | Inspect the control section and repeat with a separately validated IHC antibody if available (general IHC practice). Record the discordant compartment rather than assigning it to POMC. |
| Pituitary staining is widespread and obscures cell boundaries. | Diffuse background can prevent reliable assignment to anterior pituitary cells (general IHC practice; HPA: High in anterior pituitary cells). | Compare the matched control and inspect morphology; review blocking, antibody dilution, washes, and chromogen development (general IHC practice). The supplied evidence gives no POMC-specific dilution. |
| Extracellular material stains, but nearby cells are unclear. | Secreted POMC-related material could lie away from its producing cell, so the deposit does not establish cell of origin (UniProt P01189: secreted; HPA: RNA/protein location caveat). | Score the cellular pattern separately from extracellular deposits and compare with the anterior pituitary reference pattern (HPA: High in anterior pituitary cells; general IHC practice). |
Experiment evidence
IHC & IF images showing POMC staining
PB9039 · IHCIHC analysis of ACTH using anti-ACTH antibody (PB9039). ACTH was detected in paraffin-embedded section of mouse kidney tissues. Heat mediate
PB9039 · IHCIHC analysis of ACTH using anti-ACTH antibody (PB9039). ACTH was detected in paraffin-embedded section of rat brain tissues. Heat mediated a
HPA · Pituitary glandHuman Protein Atlas IHC, Pituitary gland. Source: https://www.proteinatlas.org/ENSG00000115138-POMC/tissue/pituitary+gland#imid_1627
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115138-POMC/tissue/cerebral+cortex#imid_18121131
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115138-POMC/tissue/colon#imid_18121049
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115138-POMC/tissue/liver#imid_18121050
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for POMC IHC & IF
🧪Run pituitary gland first: cells in the anterior pituitary should stain (HPA: High in cells in anterior pituitary). Use adipose tissue as a negative, with adipocytes expected to lack staining (HPA: Not detected in adipocytes); cells without specific signal on the pituitary slide should show counterstain only, without assigning them a cell identity from the supplied HPA row.
Positive control tissue: Pituitary gland (Cells in anterior, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for POMC; derive a cell-line control from the positive tissue's cell type (Cells in anterior) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and a host-matched rabbit IgG isotype control (PB9039 caption: rabbit primary), plus POMC knockout tissue or a validated immunizing-peptide block as a biological specificity control. For chromogenic detection, quench endogenous peroxidase and check for endogenous biotin background when using the caption’s avidin–biotin detection method (PB9039 caption: SABC and DAB).
⚠️Feasibility: A POMC-specific fixation window or fixation effect is unreported, and the selected PB9039 paraffin-section caption does not report a fixative (PB9039 caption). That caption uses heat retrieval in citrate buffer at pH 6 for 20 minutes; it does not establish retrieval dependence or whether frozen sections or IF are easier (PB9039 caption). In pituitary sections, interpret staining with the antibody’s ACTH specificity in mind because POMC is a processed, secreted precursor (PB9039 caption: anti-ACTH; UniProt P01189: secreted hormone precursor).
HPA tissue IHC evidence for POMC
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. Antibody staining consistent with GTEx and FANTOM5 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 |
|---|---|---|---|---|
| Pituitary gland | Cells in anterior | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | 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 → |
Section 3
Advanced POMC IHC Tips
Troubleshoot POMC chromogenic IHC in paraffin sections by checking retrieval, cellular context, staining controls, and the epitope detected.
How should I retrieve POMC epitopes in paraffin sections?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes (datasheet PB9039). That condition accompanies a paraffin-section image stained with 1 μg/ml catalog antibody overnight at 4°C (datasheet PB9039); the caption does not state the fixative (datasheet PB9039). If staining is weak, compare controlled changes in heating time while keeping the buffer, detection chemistry, and tissue preparation constant (standard IHC practice). Include anterior pituitary as an expected positive tissue (HPA: high in anterior pituitary cells), and assess changes in background alongside signal because stronger retrieval can expose nonspecific staining (standard IHC practice).
Could fixation explain weak or variable POMC staining?
Target-specific fixation sensitivity is unknown from the supplied evidence; the catalog image describes a paraffin section but does not state its fixative (datasheet PB9039). Record the fixative, fixation duration, tissue thickness, and processing history for each specimen before comparing staining intensity (standard IHC practice). For a controlled check, process matched specimens under defined fixation conditions, then apply the same citrate pH 6, 20-minute retrieval and antibody concentration (datasheet PB9039; standard IHC practice). Interpret any difference as an effect of those tested preparations until replicated; anterior pituitary staining provides an expected positive reference, not evidence for a particular fixation response (HPA: high in anterior pituitary cells; standard IHC practice).
Where should convincing POMC staining appear?
Assess staining in the cellular context of the section: POMC is a secreted precursor with no transmembrane segment (UniProt P01189 topology and subcellular annotation). Intracellular cytoplasmic staining can reflect material before secretion, but a fixed section alone cannot establish the precise processing compartment (UniProt P01189 processing and subcellular annotation; standard IHC interpretation). Anterior pituitary cells offer the clearest tissue reference in this evidence set (HPA: high in anterior pituitary cells). Be cautious about calling sharply outlined plasma-membrane staining specific solely from its appearance, and compare the pattern with a negative control and intact cellular morphology (UniProt P01189 topology; standard IHC practice).
Does this stain detect intact POMC or a processed peptide?
Identify the antibody's immunogen or mapped epitope before naming the stained molecule: the supplied record lists 0 isoforms, while POMC is a hormone precursor subject to cleavage (UniProt P01189 isoforms and processing). The selected image describes an anti-ACTH antibody, so its staining should not automatically be described as detection of every POMC-derived product (datasheet PB9039; UniProt P01189 function). Glycosylation at residues 71 and 91, together with other listed modifications, makes epitope position relevant to interpretation (UniProt P01189 modifications). Compare epitope documentation and appropriate peptide or antibody controls when available; chromogenic signal alone cannot distinguish precursor from a product sharing that epitope (standard IHC interpretation).
How can IF help resolve an ambiguous chromogenic pattern?
Use IF as a separate assay to examine whether signal overlaps an independently validated marker of the expected anterior pituitary cells (HPA: high in anterior pituitary cells; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before assigning faint signal to POMC (standard IF practice). POMC has no transmembrane segment and is secreted (UniProt P01189 topology and subcellular annotation); if the antibody epitope is intracellular in fixed cells, test gentle permeabilisation against an unpermeabilised control (standard IF practice). The supplied catalog evidence is paraffin-section IHC, so establish IF antibody performance and compartment assignment with IF controls before comparing it with DAB staining (datasheet PB9039; standard IF practice).
How do I separate POMC signal from chromogenic background?
First compare the stained section with a no-primary control to locate signal from secondary reagents or detection chemistry (standard IHC practice). The catalog example uses 10% goat serum block, biotinylated secondary antibody, an avidin-biotin complex, and DAB (datasheet PB9039). Check endogenous peroxidase blocking and, for this detection scheme, endogenous biotin controls when diffuse or tissue-associated color persists (datasheet PB9039; standard IHC practice). Titrate primary antibody around the reported 1 μg/ml condition while holding retrieval and development constant (datasheet PB9039; standard IHC practice). Treat signal confined to damaged edges or necrotic areas as suspect until it matches intact-cell staining and control sections (standard IHC practice).
What should I measure when comparing POMC staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and eligible intact cells before scoring, and apply the same boundaries to every section (standard IHC practice). For anterior pituitary, report the percentage of positive cells and an intensity-based H-score, with the scoring thresholds and observer method stated (HPA: high in anterior pituitary cells; standard IHC practice). Normalise positive-cell counts to all eligible cells in the sampled region, or report positive-cell density per mm² of evaluable tissue (standard IHC practice). Keep section thickness, retrieval at citrate pH 6, antibody concentration, DAB development, and image acquisition consistent; exclude folds, necrosis, and edges using prespecified rules (datasheet PB9039; standard IHC practice).
When is a POMC-positive section biologically convincing?
Give greatest weight to reproducible staining of intact anterior pituitary cells, where POMC is highly expressed (HPA: high in anterior pituitary cells). Interpret kidney staining in the selected mouse paraffin-section image as evidence of that antibody's reported staining condition, not proof that every kidney signal represents locally produced POMC (datasheet PB9039; HPA: secreted protein can differ in RNA and protein location). A predominantly crisp membrane pattern conflicts with the absence of a transmembrane segment and warrants control review (UniProt P01189 topology; standard IHC interpretation). Reject conclusions driven by section edges, necrosis, or endogenous enzyme signal until morphology and no-primary controls support specific cellular staining (standard IHC practice).
Boster reagents
Best POMC / Pro-opiomelanocortin IHC Antibodies
PB9039 has paraffin-section IHC images from mouse kidney, rat brain, and rat kidney (PB9039 image captions). Human reactivity is listed, but no human IHC image is supplied (PB9039 catalog).
Real IHC data
Anti-ACTH/POMC Antibody
Cat # PB9039
PB9039 renders with its own mouse kidney paraffin-section IHC figure (PB9039 image caption). Additional PB9039 captions show rat brain and kidney paraffin sections; its catalog lists Human, Mouse, and Rat reactivity (PB9039 image captions; PB9039 catalog).
Which to pick: For tissue IHC, choose PB9039: its captions document 1 μg/ml on paraffin sections with citrate pH 6 retrieval and DAB detection; the fixative is unreported (PB9039 image captions). For cross-species work, both PB9039 and monoclonal M00305-1 list Human, Mouse, and Rat reactivity, but only PB9039 supplies IHC images in this payload (PB9039 and M00305-1 catalog; PB9039 image captions). Neither lists IF/ICC, so this payload supports no IF/ICC antibody choice (PB9039 and M00305-1 application lists).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P01189 (COLI_HUMAN, Pro-opiomelanocortin).
- Human Protein Atlas. POMC tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. POMC subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. POMC antibody validation summary (4 antibodies).
- Hypothalamic Expression of Neuropeptide Y (NPY) and Pro-OpioMelanoCortin (POMC) in Adult Male Mice Is Affected by Chronic Exposure to Endocrine Disruptors. Metabolites 2021 — PMC8228876.
- Infrared heat treatment reduces food intake and modifies expressions of TRPV3-POMC in the dorsal medulla of obesity prone rats. International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group 2011 — PMC3583363.
- Activity of arcuate kisspeptin/neurokinin B neurons and the melanocortin system during pubertal development in female sheep†. Biology of reproduction 2025 — PMC13217381.
- Role of proopiomelanocortin neuron Stat3 in regulating arterial pressure and mediating the chronic effects of leptin. Hypertension (Dallas, Tex. : 1979) 2013 — PMC3678380.
- PubMed PMID:6274691 — UniProt-cited evidence.
- PubMed PMID:6299668 — UniProt-cited evidence.
- PubMed PMID:6314261 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for POMC (Pro-opiomelanocortin). Expected localisation Secreted (UniProt); anterior pituitary cells stain (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet PB9039). Validated IHC antibody: PB9039 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Pituitary gland (Cells in anterior); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse kidney tissues; fixative not specified (datasheet PB9039); 2. Fixation: Image fixative and duration unreported (datasheet PB9039); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Citrate pH 6 , 20 min (datasheet PB9039); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9039); 8. Primary antibody: Rabbit anti-POMC, 0.5-1μg/ml (datasheet PB9039); 9. Primary incubation: Overnight at 4 °C (datasheet PB9039). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Secreted protein may stain away from its RNA source (HPA tissue IHC); Isoform / epitope: No isoforms annotated; processing may affect epitope detection (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt P01189, HPA tissue IHC · Enhanced, HPA ICC-IF · Secreted, Datasheet PB9039, PMC8228876, PMC3583363. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/POMC/immunohistochemistry-ihc-guide.