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MICU1 / Calcium uptake protein 1, mitochondrial · IHC design guide
Design Immunohistochemistry for MICU1
Plan MICU1 chromogenic IHC on paraffin sections using the catalog antibody's IHC evidence (datasheet A01895). This guide covers the mainly cytoplasmic tissue pattern, high staining in glandular cells, and the reported low consistency between staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MICU1 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 | Mainly cytoplasmic in tissue (HPA tissue IHC); mitochondrial intermembrane space and inner membrane (UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues; high in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A01895) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA expression show low consistency (HPA tissue IHC) | |
| Regulation | No expression inducer specified (UniProt) | |
| Isoform / epitope | 6 isoforms; mature chain starts at aa 34; epitope unspecified (UniProt) |
Section 1
Recommended MICU1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A01895) with published MICU1 IHC protocols for oral, liver, and breast specimens (PMC8195654; PMC7693594; PMC8805845; PMC11190767).
| Sample | Paraffin-embedded Mouse testis tissue; fixative not specified (datasheet A01895) |
| Fixation | Image fixative and duration unreported (datasheet A01895); 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 6.0, 20min (datasheet A01895) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Mouse monoclonal (clone Mix) anti-MICU1, 1:100 recommended; image 1:200 (datasheet A01895) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MICU1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Mainly cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6.0 for the catalog antibody (datasheet: A01895). Published alternatives use heated citrate or citric-acid microwave retrieval (PMC7693594; PMC8805845).
Section 2
What Is the Expected MICU1 Staining Pattern?
MICU1 localizes to the mitochondrial intermembrane space and inner membrane, with no transmembrane segment (UniProt Q9BPX6). In paraffin section IHC, expect mainly cytoplasmic staining (HPA tissue IHC), including strong staining in selected glandular, epithelial, hematopoietic, and Purkinje cells (HPA tissue IHC: High). HPA rates the tissue staining Approved but reports low consistency between antibody staining and RNA expression (HPA tissue IHC: reliability description).
| Cytoplasmic staining is strongest in colon or duodenal glandular cells and bronchial respiratory epithelial cells (HPA tissue IHC: High). | This matches the reported cell types and intensity. At routine chromogenic resolution, a mainly cytoplasmic appearance is compatible with mitochondrial MICU1 (HPA tissue IHC; UniProt Q9BPX6 location); it need not resolve into individual mitochondria (standard IHC practice). |
| Nuclear staining dominates while cytoplasmic staining is weak or absent (HPA tissue IHC: mainly cytoplasmic; UniProt Q9BPX6 location). | A predominantly nuclear pattern conflicts with the reported localization. Check the counterstain and compare primary antibody omission and known positive tissue controls before interpreting it as MICU1 (HPA tissue IHC; standard IHC practice). |
| Adipocytes stain as intensely as neighboring cell types despite their reported Low level (HPA tissue IHC: adipocytes Low). | The discrepancy raises concern for cross-reactivity or endogenous detection activity (standard IHC practice). Low is not absent: compare cellular distribution, background, and controls before calling adipocyte staining false (HPA tissue IHC: adipocytes Low). |
| Color covers stroma, empty spaces, and multiple cell types without a clear cellular pattern (standard IHC practice). | Treat widespread diffuse signal as background until controlled. HPA describes mainly cytoplasmic cellular staining, not a uniform field-wide deposit (HPA tissue IHC). Inspect primary antibody omission controls and the detection reagents (standard IHC practice). |
| No signal appears in colon glandular cells or cerebellar Purkinje cells, both reported High (HPA tissue IHC). | A blank known-positive specimen points to a possible assay failure, though HPA patterns do not guarantee every section or antibody will stain (HPA tissue IHC: Approved; low RNA consistency). Check tissue integrity and each detection step (standard IHC practice). |
💡Expected MICU1 appearanceCall a result consistent when High HPA-listed cell types show mainly cytoplasmic staining; dominant nuclear color or uniform field-wide color warrants control review (HPA tissue IHC; UniProt Q9BPX6 location; standard IHC practice).
| Cell type and comparison tissue (HPA tissue IHC) | Colon and duodenal glandular cells, bronchial respiratory epithelium, and Purkinje cells are reported High; adipocytes are Low (HPA tissue IHC). Use those levels as comparisons, not absolute positive or negative guarantees, given low staining–RNA consistency (HPA tissue IHC). |
| Antibody evidence (HPA antibodies; HPA tissue IHC) | HPA037480 is Approved for IHC; the supplied record does not label its IHC validation Enhanced (HPA antibodies). HPA's overall tissue reliability is Approved with low consistency between staining and RNA, so corroborate unexpected patterns with controls (HPA tissue IHC; standard IHC practice). |
| Subcellular resolution (UniProt Q9BPX6; HPA subcellular) | MICU1 occupies the mitochondrial intermembrane space and inner membrane (UniProt Q9BPX6). HPA ICC-IF supports mitochondrial localization (HPA subcellular); routine chromogenic IHC may present this as mainly cytoplasmic staining (HPA tissue IHC; standard IHC practice). |
| Does IF/ICC offer a localization check? (HPA subcellular) | Yes. HPA reports supported mitochondrial ICC-IF localization and images in A-431, A-549, and U2OS; HPA057525 has Supported ICC status (HPA subcellular; HPA antibodies). These data support compartment comparison but do not validate a new IHC protocol (standard IHC practice). |
| Isoforms and epitope coverage (UniProt Q9BPX6) | UniProt lists six MICU1 isoforms and a mature chain spanning residues 34–476 (UniProt Q9BPX6). The supplied evidence gives no antibody epitope, so isoform recognition and any effect of processing on this assay remain undetermined (UniProt Q9BPX6; HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| High HPA-listed cells show no detectable color (HPA tissue IHC: High). | The assay may have failed at antibody binding or chromogenic detection; HPA intensity alone cannot identify which step failed (standard IHC practice; HPA tissue IHC). | Run a reported High tissue alongside a primary antibody omission control, then review retrieval, antibody dilution, detection reagents, and counterstain (HPA tissue IHC; standard IHC practice). |
| The slide is strongly nuclear rather than mainly cytoplasmic (HPA tissue IHC; UniProt Q9BPX6 location). | Counterstain, nonspecific binding, or detection artifact may obscure the expected compartment (standard IHC practice). | Compare with the omission control and a reported High tissue; score MICU1 only where cellular color can be distinguished from nuclear counterstain (standard IHC practice; HPA tissue IHC). |
| Adipocytes or unstructured areas look as strong as reported High cells (HPA tissue IHC: adipocytes Low). | Cross-reactivity, endogenous detection activity, or excess background is possible; Low adipocyte staining does not imply zero signal (standard IHC practice; HPA tissue IHC). | Check omission and detection controls, then compare staining in adipocytes with High glandular or epithelial cells on suitably matched sections (standard IHC practice; HPA tissue IHC). |
| Brown color spreads across the section with poor cell boundaries (standard IHC practice). | Insufficient blocking or washing, excess antibody, or endogenous detection activity can create diffuse color (standard IHC practice). | Inspect controls, adjust blocking and washing, and titrate the IHC antibody within its validated conditions; the payload supplies no MICU1-specific dilution (standard IHC practice; HPA antibodies). |
| A punctate mitochondrial pattern is not individually resolved in chromogenic IHC (UniProt Q9BPX6 location). | Mitochondria may appear as a broader cytoplasmic signal at routine light-microscope resolution (HPA tissue IHC; standard IHC practice). | Judge cellular and compartment distribution first; consult HPA's supported mitochondrial ICC-IF localization for a separate resolution check (HPA tissue IHC; HPA subcellular). |
| A tissue differs from its HPA example despite clean controls (HPA tissue IHC). | HPA reports low consistency between antibody staining and RNA expression, and its listed levels describe observed examples rather than guaranteed outcomes (HPA tissue IHC). | Record the cell type, compartment, intensity, and controls; compare another reported High tissue before assigning a biological difference (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing MICU1 staining
A01895 · IHCImmunohistochemical analysis of paraffin-embedded Mouse-testis tissue. 1, MICU1 Monoclonal Antibody (Mix) was diluted at 1:200 (4°C, overnig
A01895 · IFImmunofluorescence analysis of Hela cell. 1, c-Myc Polyclonal Antibody (red) was diluted at 1:200 (4° overnight). MICU1 Monoclonal Antibody
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000107745-MICU1/tissue/cerebral+cortex#imid_9747748
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000107745-MICU1/tissue/colon#imid_9764614
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000107745-MICU1/tissue/liver#imid_9764558
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000107745-MICU1/tissue/kidney#imid_9764564
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA057525; green = MICU1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA057525; green = MICU1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MICU1 IHC & IF
🧪Run bronchus first and score respiratory epithelial cells, which HPA rates High for MICU1 (HPA: High in bronchus respiratory epithelial cells). HPA detects MICU1 in all 45 scored tissues, so there is no validated negative tissue or internal negative cell type; the no-primary and isotype controls should show counterstain without specific chromogen in cells on the positive slide (HPA: no negative tissue rows; HPA: detected in all 45 tissues).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: MICU1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MICU1 in A-431, A-549, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and an isotype control matched to the primary antibody’s host species, immunoglobulin class and clonality; use MICU1-knockout material as a biological specificity control if available (caption: secondary-only negative control; standard IHC practice). Quench endogenous peroxidase for chromogenic detection and check bronchial inflammatory cells for residual peroxidase signal (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A01895 paraffin-section caption does not state its fixative (caption: fixative unreported). That caption uses sodium citrate retrieval at pH 6.0, >98°C for 20 minutes and a 1:200 primary dilution overnight at 4°C; it establishes one workable retrieval condition, not a requirement for all specimens (caption: retrieval and dilution). Frozen-section ease relative to paraffin IHC is unreported; IF can assess the expected mitochondrial pattern, while bronchial inflammatory cells can produce endogenous peroxidase background in chromogenic IHC (HPA: supported mitochondrial localization; standard IHC practice).
HPA tissue IHC evidence for MICU1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: MICU1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced MICU1 IHC Tips
Troubleshoot MICU1 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting chromogenic signal.
What retrieval should I try first for MICU1 in paraffin sections?
Start with heat-mediated sodium citrate retrieval at pH 6.0, above 98°C for 20 minutes (datasheet A01895). The catalog antibody was shown on paraffin-embedded mouse testis with primary antibody at 1:200 overnight at 4°C; the caption does not state a fixative (datasheet A01895). If staining is weak, compare shorter and longer heating under otherwise identical conditions, recording tissue integrity and signal in the same cell populations (standard IHC practice). A secondary-only section helps distinguish retrieval-related background from primary-dependent staining; that control was included in the caption (datasheet A01895).
How should I troubleshoot fixation-related loss of MICU1 staining?
Target-specific sensitivity of MICU1 to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not report its fixative (datasheet A01895). Record the fixative, fixation duration, tissue thickness and processing schedule for each section, then compare matched sections using the same citrate pH 6.0 retrieval and antibody conditions (datasheet A01895; standard IHC practice). Include a previously staining section in the same run to separate a new processing problem from detection failure (standard IHC practice). Avoid attributing weak signal to a particular fixation chemistry without a controlled comparison; HPA tissue staining does not establish fixation sensitivity (HPA tissue IHC).
Where should convincing MICU1 chromogenic signal appear?
Assess staining within the cytoplasm, where mitochondria reside, rather than requiring a sharply resolved mitochondrial outline in a chromogenic tissue section (UniProt Q9BPX6 localisation; standard IHC practice). MICU1 occupies the mitochondrial intermembrane space, associates with the inner membrane through EMRE/SMDT1, and is also reported at cristae junctions (UniProt Q9BPX6 localisation). HPA describes mainly cytoplasmic tissue staining, while its subcellular data support mitochondrial localisation (HPA tissue IHC; HPA subcellular). Compare signal across intact cells and adjacent structures at the same magnification; diffuse nuclear-only staining calls for control review before assigning it to MICU1 (UniProt Q9BPX6 localisation; standard IHC practice).
Could isoforms or epitope accessibility explain discordant MICU1 staining?
MICU1 has 6 listed isoforms and a processed chain spanning residues 34–476, so check the antibody’s documented immunogen or epitope before assuming every isoform is detected (UniProt Q9BPX6 processing and isoforms; standard IHC practice). The supplied caption gives no epitope, making isoform coverage for this catalog antibody unresolved (datasheet A01895). MICU1 has no transmembrane segment, but resides in the mitochondrial intermembrane space and associates with the inner membrane; these facts alone do not establish epitope accessibility after processing (UniProt Q9BPX6 topology and localisation). If sections disagree, compare matched retrieval conditions and an independently validated antibody with a known epitope where available (standard IHC practice).
How can IF help resolve an ambiguous MICU1 IHC pattern?
Use IF on separately optimised material to compare MICU1 signal with a mitochondrial marker and a marker for the expected cell population (HPA subcellular; standard IF practice). For example, HPA reports high IHC staining in bronchial respiratory epithelial cells, which can guide cell identification without establishing an IF protocol (HPA tissue IHC). Select fluorophores after inspecting tissue autofluorescence in unstained controls, and keep channels distinct enough to assess overlap (standard IF practice). Because MICU1 is in the mitochondrial intermembrane space and has no transmembrane segment, optimise permeabilisation to give antibody access to its epitope without disrupting mitochondrial structure (UniProt Q9BPX6 topology and localisation; standard IF practice).
What should I check when MICU1 staining is widespread or uneven?
Run a secondary-only control alongside the primary-stained section; this control was used for the catalog antibody’s paraffin-section image (datasheet A01895). Check endogenous peroxidase blocking, nonspecific binding, wash stringency and chromogen development time when diffuse brown signal appears (standard chromogenic IHC practice). Inspect section edges, folds and damaged areas separately from intact tissue, and adjust antibody concentration only while retaining comparable controls (standard IHC practice). HPA reports mainly cytoplasmic expression in many tissues, so widespread cytoplasmic staining is not automatically background; its tissue IHC reliability is marked Approved with low consistency between staining and RNA data (HPA tissue IHC).
How should I quantify MICU1 IHC across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis area before scoring, then record the percentage of positive cells and staining intensity within intact tissue (standard IHC scoring practice). An H-score combines intensity grades 0–3 with the percentage at each grade to yield 0–300; use the same thresholds and imaging settings across samples (standard IHC scoring practice). For cell counts, report positive cells per mm² and normalise to viable analysed area or the number of eligible cells, as appropriate (standard IHC scoring practice). Report cell type separately because HPA shows high staining in several populations, including adrenal glandular and bronchial respiratory epithelial cells (HPA tissue IHC).
How do I distinguish true MICU1 staining from an artefact?
Give greatest weight to reproducible signal in intact cell cytoplasm that is compatible with mitochondrial localisation, assessed alongside the same-run controls (UniProt Q9BPX6 localisation; standard IHC practice). Compare the stained cell type with the tissue context: HPA reports high staining in colon glandular cells and cerebellar Purkinje cells, but low staining in adipocytes (HPA tissue IHC). Treat nuclear-only signal, section-edge enhancement, necrotic-area deposits and staining retained in a secondary-only control as reasons to investigate artefact (UniProt Q9BPX6 localisation; standard IHC practice). Do not infer mitochondrial calcium uptake activity from staining intensity alone; MICU1 is a calcium sensor within the uniporter complex (UniProt Q9BPX6 function).
Boster reagents
Best MICU1 / Calcium uptake protein 1, mitochondrial IHC Antibodies
A01895 has IHC data from paraffin-embedded mouse testis and IF data from HeLa cells and human appendix (A01895 image captions). Its listed reactivity is human, mouse and rat (A01895 catalog).
Real IHC data
Anti-MICU1/Cbara1 Monoclonal Antibody
Cat # A01895
A01895 is the SKU with a rendered card; its IHC figure shows paraffin-embedded mouse testis (A01895 IHC caption). Its IF images show HeLa cells and human appendix (A01895 IF captions).
Which to pick: Choose A01895 for tissue IHC because its own figure shows paraffin-embedded mouse testis; the fixative is unreported (A01895 IHC caption). For IF/ICC, A01895 is the choice because IF is listed and cell imaging is shown (A01895 applications; A01895 IF caption). For cross-species work, A01895 and A01895-3 both list human, mouse and rat reactivity, but the supplied IHC figure establishes tissue staining only for A01895 in mouse testis (catalog reactivity; A01895 IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BPX6 (MICU1_HUMAN, Calcium uptake protein 1, mitochondrial).
- Human Protein Atlas. MICU1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MICU1 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. MICU1 antibody validation summary (2 antibodies).
- MCU That Is Transcriptionally Regulated by Nrf2 Augments Malignant Biological Behaviors in Oral Squamous Cell Carcinoma Cells. BioMed research international 2021 — PMC8195654.
- A Novel Biomarker Driving Poor-Prognosis Liver Cancer: Overexpression of the Mitochondrial Calcium Gatekeepers. Biomedicines 2020 — PMC7693594.
- Dihydroartemisinin represses oral squamous cell carcinoma progression through downregulating mitochondrial calcium uniporter. Bioengineered 2022 — PMC8805845.
- Multi-omic profiling of breast tumor microenvironment uncovers a role of mitochondrial calcium gatekeepers. Journal of Cancer 2024 — PMC11190767.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MICU1 (Calcium uptake protein 1, mitochondrial). Expected localisation Mainly cytoplasmic in tissue (HPA tissue IHC); mitochondrial intermembrane space and inner membrane (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A01895). Validated IHC antibody: A01895 at 1:100 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded Mouse testis tissue; fixative not specified (datasheet A01895); 2. Fixation: Image fixative and duration unreported (datasheet A01895); 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 6.0 , 20min (datasheet A01895); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Mouse monoclonal (clone Mix) anti-MICU1, 1:100 recommended; image 1:200 (datasheet A01895); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA expression show low consistency (HPA tissue IHC); Isoform / epitope: 6 isoforms; mature chain starts at aa 34 ; epitope unspecified (UniProt); Regulation: No expression inducer specified (UniProt). Evidence: UniProt Q9BPX6, HPA tissue IHC · Approved, HPA ICC-IF · mitochondria., Datasheet A01895, PMC8195654, PMC7693594. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MICU1/immunohistochemistry-ihc-guide.