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- Table of Contents
Plan MDH2 paraffin-section IHC around the granular cytoplasmic staining reported across tissues (HPA tissue IHC). Compare cell type and staining intensity while treating mitochondrial matrix localization as a molecular expectation (UniProt).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Granular cytoplasm (HPA tissue IHC); mitochondrial matrix (UniProt) | |
| Staining pattern | Granular cytoplasmic staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04803-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
| Fixation | Keep formalin fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Broad tissue staining makes negative controls harder to select (HPA tissue IHC) | |
| Regulation | RNA enhanced in skeletal muscle and tongue (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; mature chain spans residues 25–338; epitope coverage unknown (UniProt) |
The catalog antibody’s IHC-P protocol is followed by three published MDH2 IHC methods with usable preparation or staining details (PMC10868282; PMC8586664; PMC5300292).
| Sample | Paraffin-embedded human laryngeal carcinoma tissue; fixative not specified (datasheet A04803-2) |
| Fixation | Image fixative and duration unreported (datasheet A04803-2); 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 A04803-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04803-2) |
| Primary antibody | Rabbit anti-MDH2, 1-2 μg/ml (datasheet A04803-2) |
| Primary incubation | Overnight at 4 °C (datasheet A04803-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04803-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MDH2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in all tissues. No signal in the no-primary control. |
MDH2 is a mitochondrial matrix protein with no transmembrane segment (UniProt P40926). In paraffin-section IHC, expect granular cytoplasmic staining across tissues, with high staining in several glandular and neural cell populations (HPA: tissue IHC). HPA rates the tissue pattern Enhanced, citing high consistency between antibody staining and RNA expression data (HPA: tissue IHC).
| Granular cytoplasmic staining in adrenal glandular cells or bronchial respiratory epithelium. | This matches high staining reported in those cells (HPA: tissue IHC). Granules are consistent with mitochondrial localisation (UniProt P40926; HPA: subcellular). |
| Predominantly nuclear or crisp plasma-membrane staining. | This conflicts with the mitochondrial matrix assignment (UniProt P40926). Treat it as possible nonspecific staining or an imaging artefact; compartment alone cannot identify the cause. |
| Strong staining in smooth muscle cells while expected glandular cells stain weakly. | HPA reports low staining in smooth muscle cells and high staining in several glandular populations (HPA: tissue IHC). Consider cross-reactivity or endogenous detection activity; neither is proven by this pattern. |
| Diffuse chromogen throughout cells, tissue spaces, or the whole section, obscuring granules. | The distribution is difficult to reconcile with HPA's granular cytoplasmic profile (HPA: tissue IHC). Background may arise from detection or washing conditions (general IHC practice). |
| No detectable staining in a section expected to contain high-expressing cells. | A blank adrenal glandular or bronchial epithelial population conflicts with HPA's high staining reports (HPA: tissue IHC). Check assay performance before interpreting it as absent MDH2. |
| Subcellular assignment and topology | MDH2 is assigned to the mitochondrial matrix and has no transmembrane segment (UniProt P40926). Interpret IHC granules as a compartment-compatible pattern, not proof that each granule is a mitochondrion. |
| Tissue and cell context | HPA describes granular cytoplasmic expression in all tissues, with high staining in listed glandular, respiratory epithelial, glial, and neuronal populations and low staining in smooth muscle cells (HPA: tissue IHC). Compare cells within their tissue context. |
| Strength of pattern evidence | The HPA tissue profile is rated Enhanced for agreement between antibody staining and RNA expression; four listed antibodies have Enhanced IHC status (HPA: tissue IHC; HPA: antibodies). These ratings support the reported pattern, but do not validate an unlisted catalog antibody. |
| Protein processing and isoforms | UniProt annotates a mature chain spanning residues 25–338 and two isoforms (UniProt P40926). Without an epitope map for the antibody used, this record cannot establish which forms it detects or predict a form-specific tissue pattern. |
| Fixation and retrieval evidence | The supplied UniProt and HPA records do not report MDH2-specific fixation sensitivity or antigen retrieval conditions. Treat retrieval optimisation as a general paraffin-IHC workflow decision, not a target-specific prediction. |
| IF/ICC Q&A: what location should fluorescence show? | Mitochondrial localisation is reported by HPA ICC-IF and agrees with the UniProt matrix assignment (HPA: subcellular; UniProt P40926). This supports an IF localisation check; it does not establish an IF protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| High-staining reference cells show no chromogen. | Possible assay failure; HPA reports high staining in adrenal glandular and bronchial respiratory epithelial cells (HPA: tissue IHC). | Verify that the expected cells are present, then check primary-antibody and detection controls; review retrieval and dilution using the antibody's IHC-P instructions (general IHC practice). |
| Staining is mostly nuclear or outlines cell membranes. | That distribution conflicts with MDH2's mitochondrial matrix location and HPA's granular cytoplasmic IHC profile (UniProt P40926; HPA: tissue IHC). | Review morphology and counterstain, compare with a primary-omission control, and reassess antibody specificity if the pattern persists (general IHC practice). |
| Broad diffuse colour masks intracellular granules. | Diffuse deposition can reflect excess detection background or inadequate washing (general IHC practice); it cannot be scored against HPA's granular profile (HPA: tissue IHC). | Inspect the primary-omission control, washing, blocking, and chromogen development; adjust the workflow according to the detection system's instructions (general IHC practice). |
| Smooth muscle cells appear as strongly stained as nearby expected high-staining cells. | HPA lists smooth muscle cells as low, while several other cell populations are high (HPA: tissue IHC). Cross-reactivity or endogenous detection activity is possible. | Confirm cell identity on the counterstained section, inspect a primary-omission control, and compare with an independently validated antibody if available (general IHC practice). |
| Some tissue compartments stain weakly despite strong staining elsewhere. | Intensity varies by cell population: HPA reports high staining in named populations and low staining in smooth muscle cells (HPA: tissue IHC). | Score identified cell populations separately and compare each with its HPA entry; avoid treating a low-staining compartment alone as assay failure (HPA: tissue IHC). |
| IF/ICC shows fluorescence outside the expected mitochondrial distribution. | A nonmitochondrial pattern conflicts with HPA's ICC-IF location and UniProt's matrix assignment (HPA: subcellular; UniProt P40926). | Use the separate IF/ICC guide for its controls and imaging checks; this IHC section supplies a location expectation, not an IF protocol. |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: MDH2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Use compartment, cell type and control sections to assess MDH2 staining in paraffin-section IHC; the catalog image provides a starting retrieval and detection workflow.
The catalog shows MDH2 IHC staining in human paraffin sections and IF/ICC staining in human cells (catalog image captions). Listed antibody reactivity extends to Mouse and Rat (catalog reactivity).
A04803-2 has IHC images from human laryngeal carcinoma, Hashimoto thyroiditis, and liver cancer paraffin sections, plus IF/ICC in MCF-7 cells; M04803-1 has IHC images from human breast, cervical, and squamous cell carcinoma paraffin sections, plus IF/ICC in HeLa cells (catalog image captions). A04803-1 has an IHC image from formalin-fixed, paraffin-embedded human lung carcinoma (catalog IHC image caption).
Which to pick: For tissue IHC, A04803-2 offers a rabbit polyclonal antibody with human paraffin-section images, while M04803-1 offers a mouse monoclonal with human paraffin-section images; both captions specify EDTA retrieval at pH 8.0, but leave fixation unreported (catalog antibody details and IHC image captions). A04803-1 is another tissue IHC choice with an image from paraffin-section, paraffin-embedded human lung carcinoma (catalog IHC image caption). For IF/ICC, choose A04803-2 or M04803-1, each listed for IF/ICC and imaged in human cells; for cross-species IHC, A04803-2 lists Human, Mouse, and Rat reactivity, though its tissue IHC images show Human only (catalog applications, reactivity, and image captions). The selected A04803-2 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A04803-2).