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- Table of Contents
This guide uses high-staining hepatocytes and adipocytes with no detected staining as IHC tissue controls (HPA tissue IHC). Interpret the reported cytoplasmic pattern alongside MCCC2’s mitochondrial matrix location, and account for medium staining/RNA consistency when assessing results (HPA tissue IHC; UniProt).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Cytoplasmic tissue staining (HPA tissue IHC); mitochondrial matrix location (UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues; high in hepatocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06515-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining/RNA consistency is medium; verify tissue signal (HPA tissue IHC) | |
| Regulation | Liver-enhanced RNA expression (HPA tissue RNA) | |
| Isoform / epitope | Two isoforms; mature chain spans residues 23–563; epitope impact unknown (UniProt) |
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet: A06515-2). Published IHC protocols for prostate, kidney, and colorectal tissue follow (PMC7395692; PMC13347088; PMC10571261).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A06515-2) |
| Fixation | Image fixative and duration unreported (datasheet A06515-2); 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 A06515-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06515-2) |
| Primary antibody | Rabbit anti-MCCC2, 2-5 μg/ml (datasheet A06515-2) |
| Primary incubation | Overnight at 4 °C (datasheet A06515-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06515-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MCCC2-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
MCCC2 is a mitochondrial matrix protein with no transmembrane segment (UniProt Q9HCC0: location and topology). In paraffin sections, expect cytoplasmic staining in glandular cells, hepatocytes, cardiomyocytes, and kidney collecting ducts (HPA: tissue IHC). HPA describes cytoplasmic expression in most tissues; its tissue profile is Enhanced, with medium agreement between staining and RNA data and external verification pending (HPA: tissue IHC reliability).
| Cytoplasmic staining in liver hepatocytes and colon or duodenum glandular cells. | This matches HPA High protein staining in those cells (HPA: tissue IHC). Read a mitochondrial matrix protein's chromogenic signal within the cytoplasm; IHC alone does not prove organelle identity (UniProt Q9HCC0: location; standard IHC interpretation). |
| Predominantly nuclear, membranous, or extracellular staining in otherwise positive cells. | These compartments disagree with the mitochondrial matrix assignment and lack support in the HPA cytoplasmic profile (UniProt Q9HCC0: location and topology; HPA: tissue IHC). Reassess specificity and detection controls before scoring them as MCCC2 (standard IHC practice). |
| Strong staining in adipocytes, skeletal myocytes, or smooth muscle cells. | HPA reports MCCC2 as Not detected in these cell types (HPA: tissue IHC). Unexpected signal may reflect cross-reactivity or endogenous detection activity; check controls before calling it target expression (standard IHC practice). |
| Diffuse color across cells and tissue, including areas expected to be negative. | An indiscriminate signal does not match HPA's cell-specific High and Not detected observations (HPA: tissue IHC). Background from the detection system or insufficient blocking is possible; compare controls and staining conditions (standard IHC practice). |
| No signal in liver hepatocytes or heart cardiomyocytes. | Both are High in HPA tissue IHC, so an absent signal warrants a technical check (HPA: tissue IHC). Check the IHC-validated antibody's documented procedure and positive control before interpreting a sample as negative (standard IHC practice). |
| Compartment and topology | MCCC2 is assigned to the mitochondrial matrix and has no transmembrane segment (UniProt Q9HCC0: location and topology). Judge chromogenic IHC chiefly by cytoplasmic distribution and the stained cell type (HPA: tissue IHC; standard IHC interpretation). |
| Choice of tissue control | Liver hepatocytes, heart cardiomyocytes, and appendix glandular cells are High; adipocytes and skeletal myocytes are Not detected (HPA: tissue IHC). These observations offer contrasting controls, but HPA's Enhanced profile awaits external verification (HPA: reliability). |
| RNA and protein evidence | Liver is tissue enhanced at the RNA level, while hepatocytes are High by tissue IHC (HPA: RNA specificity and tissue IHC). Use the observed protein pattern for slide interpretation; RNA enrichment does not specify chromogenic intensity in every liver cell (standard IHC interpretation). |
| Antibody validation | HPA038300, HPA038301, and HPA061546 each have Enhanced IHC status (HPA: antibody validation). This supports the reported pattern, while the tissue profile's medium RNA agreement and pending external verification remain relevant limits (HPA: tissue IHC reliability). |
| Sequence and processing | UniProt lists two isoforms and a processed chain spanning residues 23–563 (UniProt Q9HCC0: isoforms and processing). No epitope position is supplied, so these records cannot establish which isoforms the IHC-validated antibody detects (UniProt Q9HCC0; supplied antibody data). |
| IF/ICC Q&A | Q: What localisation should IF/ICC show? A: Mitochondrial signal, as reported in HPA's enhanced subcellular data (HPA: ICC-IF). That observation supports the localisation expectation; it does not establish an IHC-P retrieval or detection setting (HPA: ICC-IF; standard assay distinction). |
| Situation | Likely cause | Next action |
|---|---|---|
| Liver control is blank. | HPA reports High hepatocyte staining, making a technical failure plausible (HPA: tissue IHC). | Check the IHC-validated antibody's IHC-P procedure, dilution, retrieval, detection reagents, and control section (standard IHC practice); no target-specific retrieval condition is supplied. |
| Color is widespread, including adipocytes. | Adipocytes are Not detected in HPA; excess chromogen or detection background is possible (HPA: tissue IHC; standard IHC practice). | Compare a no-primary control, review blocking and detection, and adjust development time using the documented IHC-P procedure (standard IHC practice). |
| Strong nuclear signal dominates. | Nuclear staining conflicts with the matrix location and HPA cytoplasmic profile (UniProt Q9HCC0: location; HPA: tissue IHC). | Check localisation in a known-positive tissue and compare the no-primary control before scoring; review antibody specificity if nuclear signal persists (standard IHC practice). |
| Smooth muscle stains as strongly as adjacent glands. | HPA lists smooth muscle cells as Not detected but several glandular cell populations as High (HPA: tissue IHC). | Confirm cell identity on the counterstained section and compare detection controls; treat persistent smooth muscle staining as unverified (standard IHC practice). |
| Only faint staining appears in hippocampal neurons. | HPA reports Low staining in hippocampal neuronal cells, so faint signal may fit the reference (HPA: tissue IHC). | Check a High control, such as liver hepatocytes, before changing staining conditions or calling the run insensitive (HPA: tissue IHC; standard IHC practice). |
| Expected cytoplasmic staining varies between sections. | The supplied sources establish tissue patterns but give no MCCC2-specific fixation sensitivity or retrieval setting (HPA: tissue IHC; UniProt Q9HCC0). | Compare section and run controls, then follow the IHC-validated antibody's documented IHC-P procedure consistently (standard IHC practice); avoid assigning the variation to MCCC2-specific fixation effects. |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Use compartment, cell type and control tissue together when optimising MCCC2 staining; the paraffin-section conditions below come from the selected antibody’s tissue image (datasheet A06515-2).
Both anti-MCCC2 antibodies have human paraffin-section IHC images; A06515-2 also lists mouse and rat reactivity, while A06515-1 lists rat reactivity (catalog image captions and reactivity).
A06515-2 will render with an IHC image from a paraffin-embedded human breast cancer section; its listed IHC reactivity is human, mouse and rat (A06515-2 image caption and catalog reactivity). A06515-1 will render with an IHC-P image from formalin-fixed, paraffin-embedded human hepatocarcinoma; its listed reactivity is human and rat (A06515-1 image caption and catalog reactivity).
Which to pick: For tissue IHC, choose A06515-2 if its documented EDTA retrieval at pH 8.0 and 2 μg/ml starting concentration suit the experiment; its paraffin-section caption does not report a fixative (A06515-2 image caption). Choose A06515-1 for a documented paraffin-section, paraffin-embedded human tissue example, or A06515-2 when mouse reactivity is needed (A06515-1 image caption; catalog reactivity). Neither SKU lists IF/ICC validation or an IF dilution, so neither is a validated IF/ICC choice here (catalog applications and dilutions). The selected A06515-2 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A06515-2).