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MIA3 / Transport and Golgi organization protein 1 homolog · IHC design guide
Design Immunohistochemistry for MIA3
Plan MIA3 IHC in paraffin sections using the expected cytoplasmic tissue pattern and ER exit site localisation (HPA tissue IHC; UniProt). Compare staining with rectal glandular cells, which show high expression, and adipocytes, where MIA3 is not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MIA3 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); ER exit sites expected (UniProt) | |
| Staining pattern | Cytoplasmic staining in cells of most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06158-1) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+3 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 | Signal varies by cell type; adipocytes are not detected (HPA tissue IHC) | |
| Regulation | Down-regulated in melanoma tissue (UniProt) | |
| Isoform / epitope | 4 isoforms; check whether the epitope is lumenal or cytoplasmic (UniProt) |
Section 1
Recommended MIA3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A06158-1) with published MIA3 staining methods for clinical sections (PMC5576312) and tumour xenografts (PMC11455670).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A06158-1) |
| Fixation | Image fixative and duration unreported (datasheet A06158-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A06158-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06158-1) |
| Primary antibody | Rabbit anti-MIA3, 2-5 μg/ml (datasheet A06158-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06158-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06158-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MIA3-positive staining in cells in molecular layer of cerebellum (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 retrieval at pH 8.0 (datasheet A06158-1); a published alternative uses 10 mM citrate with heat-induced retrieval (PMC5576312).
Section 2
What Is the Expected MIA3 Staining Pattern?
MIA3 is an endoplasmic reticulum (ER) membrane protein enriched at ER exit sites; its large N-terminal region is lumenal and its C-terminal region is cytoplasmic (UniProt Q5JRA6 topology and subcellular location). In paraffin-section IHC, expect predominantly cytoplasmic staining in the cell populations reported by HPA, including rectal glandular cells and cerebral cortical neurons (HPA tissue IHC). HPA rates its tissue staining profile Supported, with consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining in rectal glandular cells or cerebral cortical neurons. | This fits HPA's High staining in these cells and its broader cytoplasmic tissue profile (HPA tissue IHC). Compare signal within the identified cells; do not assume every neighboring cell should stain equally. |
| Predominantly nuclear staining or staining confined to the extracellular space, without convincing cytoplasmic signal. | This does not fit the ER membrane location (UniProt Q5JRA6 subcellular location) or the cytoplasmic tissue profile (HPA tissue IHC). Treat it as a localization concern and review controls and morphology before scoring it as MIA3. |
| Strong staining in adipocytes, alveolar cells, skeletal muscle myocytes or chondrocytes. | HPA reports MIA3 as Not detected in those respective cell populations (HPA tissue IHC). Unexpected staining warrants a check for antibody cross-reactivity or endogenous detection activity; tissue-level absence is not proof that every positive-looking pixel is an artefact. |
| Diffuse color across cells, stroma and tissue edges, with little distinction between cell populations. | A widespread haze is difficult to reconcile with HPA's cell-specific levels and cytoplasmic profile (HPA tissue IHC). In chromogenic IHC, nonspecific antibody binding or detection background can obscure a real signal (general IHC practice). |
| No staining in a well-preserved rectal glandular or cerebral cortical neuronal positive-control area. | HPA reports High staining in those cells, so a blank control challenges the run before it supports biological absence in a test specimen (HPA tissue IHC). Check that the intended cells are present and that the staining and detection steps worked. |
💡Expected MIA3 appearanceCall a result convincing when identifiable High-expression cells show clear cytoplasmic staining, such as rectal glandular cells or cerebral cortical neurons (HPA tissue IHC), consistent with MIA3's ER membrane location (UniProt Q5JRA6); isolated nuclear color or broad cell-free haze is a localization or background warning (general IHC practice).
| Cell population and tissue choice | HPA reports High staining in rectal glandular cells, cerebral cortical neurons, nasopharyngeal respiratory epithelium and pachytene spermatocytes, but Not detected in adipocytes and alveolar cells (HPA tissue IHC). Use the named cell population when interpreting a section; an organ name alone is too coarse. |
| Expected compartment | MIA3 is an ER membrane protein at ER exit sites, with one transmembrane segment at residues 1177–1197 (UniProt Q5JRA6 topology and subcellular location). HPA describes tissue IHC as cytoplasmic in most tissues (HPA tissue IHC). Routine chromogenic sections need not resolve individual exit sites. |
| Antibody evidence | HPA rates the tissue profile Supported and lists IHC as Supported for HPA055922 and HPA056816 (HPA tissue IHC; HPA antibodies). These summaries support the reported pattern but do not validate every antibody or explain an unexpected pattern from the antibody used in a separate experiment. |
| Protein forms and epitope interpretation | UniProt lists four isoforms, a cleaved signal peptide at residues 1–22 and two glycosylation sites at residues 246 and 589 (UniProt Q5JRA6 isoforms, processing and glycosylation). Without an antibody's mapped epitope, these facts cannot establish which forms it stains or predict an antigen-retrieval response. |
| IF/ICC Q: Should vesicular signal match the IHC appearance? | A: HPA calls vesicles the enhanced main location in ICC-IF images from A-549, U-251MG and U2OS, while its tissue IHC summary says cytoplasmic expression (HPA subcellular ICC-IF; HPA tissue IHC). Interpret each assay at its own resolution; the IF/ICC guide covers that application. |
| Fixation sensitivity | Target-specific fixation effects and antigen-retrieval requirements are unreported in the supplied UniProt and HPA records (UniProt Q5JRA6; HPA tissue IHC). Any retrieval adjustment is a general IHC workflow decision, not an evidence-based prediction of altered MIA3 staining. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control cells are blank. | The expected High population may be absent from the viewed area (HPA tissue IHC), or a staining or detection step may have failed (general IHC practice). | Locate the named cells on the counterstained section, then review primary-antibody, detection and chromogen steps with an appropriate run control (general IHC practice). |
| Only nuclei appear positive. | Nuclear-only color conflicts with the ER membrane location (UniProt Q5JRA6 subcellular location) and HPA's cytoplasmic IHC profile (HPA tissue IHC). | Check morphology and compare with a primary-omission or other appropriate negative control for nonspecific detection (general IHC practice); avoid scoring nuclear color alone as MIA3. |
| Adipocytes or alveolar cells stain strongly. | These are reported as Not detected by HPA (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Confirm cell identity, inspect the negative control and compare the same run with a named High population (HPA tissue IHC; general IHC practice). |
| Color covers stroma and tissue edges. | Broad background may reflect nonspecific binding or detection activity (general IHC practice); it obscures HPA's cytoplasmic, cell-associated profile (HPA tissue IHC). | Inspect a primary-omission control and review blocking, washing and chromogen development for the assay (general IHC practice). |
| A weak test section seems negative. | Some named populations are Low, including hepatocytes and duodenal glandular cells, whereas others are High (HPA tissue IHC). A weak result alone cannot establish assay failure or true absence. | Score the relevant cell population and compare it with a High-expression control processed in the same run (HPA tissue IHC; general IHC practice). |
| A tissue IHC image looks less punctate than an ICC-IF image. | HPA describes cytoplasmic tissue IHC and enhanced vesicular ICC-IF localization (HPA tissue IHC; HPA subcellular ICC-IF); chromogenic section detail differs from fluorescence microscopy (general IHC/IF practice). | Judge paraffin-section IHC against its cytoplasmic cell-pattern expectation and controls (HPA tissue IHC; general IHC practice); use the separate IF/ICC guide for fluorescence interpretation. |
Experiment evidence
IHC & IF images showing MIA3 staining
A06158-1 · IHCIHC analysis of MIA3 using anti-MIA3 antibody (A06158-1). MIA3 was detected in a paraffin-embedded section of human colorectal adenocarcinom
A06158-1 · IHCIHC analysis of MIA3 using anti-MIA3 antibody (A06158-1). MIA3 was detected in a paraffin-embedded section of human liver cancer tissue. Hea
A06158-1 · IFIF analysis of MIA3 using anti-MIA3 antibody (A06158-1). MIA3 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000154305-MIA3/tissue/cerebral+cortex#imid_16180919
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000154305-MIA3/tissue/colon#imid_16180837
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000154305-MIA3/tissue/liver#imid_16180838
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000154305-MIA3/tissue/pancreas#imid_16180789
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA055922; green = MIA3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA055922; green = MIA3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MIA3 IHC & IF
🧪Run rectum first and score MIA3 staining in glandular cells (HPA: High in rectal glandular cells); use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes). On the rectal slide, compare non-glandular cells with the no-primary control for background staining; their MIA3-negative status is unverified.
Positive control tissue: Cerebellum (Cells in molecular layer, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MIA3 in A-549, U-251MG, U2OS, with annotated localisation: Vesicles (enhanced) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched rabbit IgG isotype control, and an MIA3 knockout specimen if available (A06158-1 tissue-IHC caption: rabbit primary antibody). Quench endogenous peroxidase and check the rectal slide for background DAB staining (A06158-1 tissue-IHC caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the fixative is unreported in the A06158-1 tissue-IHC caption. Heat retrieval in EDTA at pH 8.0 preceded staining, but whether retrieval is required has not been established (A06158-1 tissue-IHC caption). Frozen sections and IF are not shown to be easier; in rectal glands, distinguish intracellular staining from deposits in gland lumens (HPA: High in rectal glandular cells; standard IHC practice).
HPA tissue IHC evidence for MIA3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High consistency between antibody staining and RNA expression data. Paired antibodies with high similarity supports the protein expression profile.). 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 |
|---|---|---|---|---|
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Rectum | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Pachytene spermatocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Chondrocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MIA3 IHC Tips
Use the catalog antibody’s paraffin section evidence to troubleshoot MIA3 chromogenic IHC, then check whether staining fits its reported cellular distribution.
Which retrieval condition should I start with for weak MIA3 staining in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A06158-1). The selected paraffin section example used that condition before incubation with 2 µg/ml primary antibody overnight at 4°C (datasheet A06158-1). If staining is weak, compare retrieval duration on matched sections while holding antibody concentration, detection and development constant (standard IHC practice). Check that tissue remains intact and that positive cells show predominantly cytoplasmic staining, consistent with the supported tissue profile (HPA: cytoplasmic expression in most tissues). Do not infer an optimal retrieval duration from the caption, which reports the buffer and pH but no heating time (datasheet A06158-1).
Could fixation explain weak or uneven MIA3 staining?
MIA3-specific fixation sensitivity is unknown from the supplied evidence; the selected paraffin section caption does not state a fixative (datasheet A06158-1). Record each specimen’s fixative and fixation time, then compare sections processed and stained together so preanalytic differences are visible (standard IHC practice). Examine morphology and staining across the section before changing retrieval or primary antibody concentration, because uneven processing can complicate interpretation (standard IHC practice). Use the documented EDTA pH 8.0 retrieval and 2 µg/ml primary antibody as starting conditions for the comparison (datasheet A06158-1). Do not assign a MIA3-specific fixation effect without a direct, controlled comparison (standard IHC practice).
Where should convincing MIA3 chromogenic staining appear?
Expect predominantly cytoplasmic staining in tissue sections, while recognising that conventional chromogenic IHC may not resolve individual ER exit sites (HPA: cytoplasmic expression in most tissues; UniProt Q5JRA6: ER exit site localisation). MIA3 is an ER membrane protein enriched at ER exit sites, and SEC16A supports its localisation there (UniProt Q5JRA6: subcellular location). Its transmembrane segment spans residues 1177–1197, with a lumenal region at 23–1144 and cytoplasmic region at 1198–1907 (UniProt Q5JRA6 topology). Treat isolated nuclear or extracellular chromogen as a reason to recheck specificity, morphology and background controls (standard IHC practice). Compare the candidate signal with neighbouring cells and a matched negative control before assigning a compartment (standard IHC practice).
How can epitope location and isoforms change the MIA3 IHC result?
First establish which sequence the catalog antibody recognises; the supplied caption does not identify its epitope (datasheet A06158-1). MIA3 has 4 reported isoforms, so staining cannot establish which isoform is present unless antibody recognition across isoforms is known (UniProt Q5JRA6: isoforms). Its lumenal region includes glycosylation sites at residues 246 and 589, while the cytoplasmic region begins at 1198 (UniProt Q5JRA6 topology and glycosylation). Retrieval may expose epitopes differently, but these annotations alone do not predict this antibody’s staining response (standard IHC practice). Compare an independently mapped antibody or an orthogonal expression measure if isoform or epitope specificity drives the conclusion (standard IHC practice).
How should I adapt the IHC interpretation when checking MIA3 by IF?
Treat IF as a separate assay: the selected antibody evidence describes chromogenic staining in a paraffin section, with no IF procedure specified (datasheet A06158-1). Multiplex MIA3 with a marker for the cell type under study, then assess whether its signal falls within those cells and shows a plausible intracellular distribution (standard IF practice; UniProt Q5JRA6: ER exit site localisation). Choose fluorophores and imaging channels after measuring tissue autofluorescence with appropriate controls (standard IF practice). Set permeabilisation for the antibody’s mapped epitope: residues 23–1144 are lumenal, whereas 1198–1907 are cytoplasmic; the supplied caption does not map this antibody’s epitope (UniProt Q5JRA6 topology; datasheet A06158-1). HPA reports an enhanced vesicular IF location, which can guide comparison without establishing equivalence to chromogenic tissue staining (HPA subcellular: vesicles).
What should I check when MIA3 DAB staining is diffuse or patchy?
Begin with a section lacking primary antibody and inspect DAB deposition, tissue edges and damaged areas alongside the stained section (standard IHC practice). Block endogenous peroxidase before HRP detection and use an appropriate protein or serum block to assess nonspecific staining (standard IHC practice). The selected example used 10% goat serum, 2 µg/ml primary antibody overnight at 4°C, and an HRP-linked secondary before DAB development (datasheet A06158-1). Adjust one variable at a time, especially blocking, primary concentration or DAB development time, while keeping matched sections together (standard IHC practice). Broad cytoplasmic expression is reported, so widespread staining alone does not establish background or specificity (HPA: cytoplasmic expression in most tissues).
How should I quantify MIA3 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then use the same criteria for every section (standard IHC practice). Report the percentage of positive cells with an intensity-based H-score, or report positive cell density per mm² when counts suit the tissue layout (standard IHC practice). Normalise counts to viable tissue area or the number of evaluable target cells, and exclude folds, necrosis and edge artefacts consistently (standard IHC practice). Record retrieval, antibody concentration and DAB development conditions because these can shift measured intensity between runs (standard IHC practice). Keep cell types separate where possible: HPA reports high signal in rectal glandular cells but no detected signal in lung alveolar cells (HPA tissue IHC).
How do I distinguish true MIA3 staining from an artefact?
A plausible result is intracellular, predominantly cytoplasmic staining in intact cells, consistent with tissue IHC and MIA3’s ER membrane localisation (HPA: cytoplasmic expression in most tissues; UniProt Q5JRA6: subcellular location). Check whether staining follows the expected cell population; HPA reports high signal in rectal glandular cells and no detected signal in adipocytes (HPA tissue IHC). Treat isolated nuclear, extracellular or sharply edge-restricted DAB, and signal concentrated in necrotic areas, as prompts to inspect morphology and controls (standard IHC practice). A no-primary section helps reveal endogenous enzyme activity or detection background, while an independently validated measure can strengthen a biological claim (standard IHC practice). The selected colorectal adenocarcinoma image demonstrates staining under its stated conditions, without establishing specificity for every specimen (datasheet A06158-1).
Boster reagents
Best MIA3 / Transport and Golgi organization protein 1 homolog IHC Antibodies
A06158-1 has IHC images from human paraffin sections and an IF/ICC image from U2OS cells (catalog image captions); its listed reactivity is human (datasheet).
Real IHC data
Anti-MIA3 Antibody ®
Cat # A06158-1
A06158-1 will render with human paraffin-section IHC data from colorectal adenocarcinoma, liver cancer, and testicular germ cell tumor (catalog IHC captions). The same SKU lists IF/ICC and has a U2OS-cell IF image (datasheet; catalog IF caption).
Which to pick: Choose A06158-1 for human tissue IHC on paraffin sections, as shown in its own IHC captions (catalog IHC captions). Choose A06158-1 for IF/ICC because both applications are listed and its IF caption shows U2OS-cell staining (datasheet; catalog IF caption). No cross-species or clone-based choice is supported: only human reactivity is listed, clonality is unreported, and the paraffin-section IHC captions do not report the fixative (datasheet; catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q5JRA6 (TGO1_HUMAN, Transport and Golgi organization protein 1 homolog).
- Human Protein Atlas. MIA3 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. MIA3 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. MIA3 antibody validation summary (2 antibodies).
- MicroRNA-222 influences migration and invasion through MIA3 in colorectal cancer. Cancer cell international 2017 — PMC5576312.
- Evidence for genetic association between chromosome 1q loci and predisposition to colorectal neoplasia. British journal of cancer 2017 — PMC5589990.
- MIA3 promotes the degradation of GSH (glutathione) by binding to CHAC1, thereby promoting the progression of hepatocellular carcinoma. Molecular and cellular biochemistry 2024 — PMC11455670.
- Global defects in collagen secretion in a Mia3/TANGO1 knockout mouse. The Journal of cell biology 2011 — PMC3105544.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MIA3 (Transport and Golgi organization protein 1 homolog). Expected localisation Cytoplasmic staining (HPA tissue IHC); ER exit sites expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06158-1). Validated IHC antibody: A06158-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Cells in molecular layer); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A06158-1); 2. Fixation: Image fixative and duration unreported (datasheet A06158-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A06158-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06158-1); 8. Primary antibody: Rabbit anti-MIA3, 2-5 μg/ml (datasheet A06158-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06158-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Signal varies by cell type; adipocytes are not detected (HPA tissue IHC); Isoform / epitope: 4 isoforms; check whether the epitope is lumenal or cytoplasmic (UniProt); Regulation: Down-regulated in melanoma tissue (UniProt). Evidence: UniProt Q5JRA6, HPA tissue IHC · Supported, HPA ICC-IF · vesicles., Datasheet A06158-1, PMC5576312, PMC11455670. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MIA3/immunohistochemistry-ihc-guide.