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TOMM34 / Mitochondrial import receptor subunit TOM34 · IHC design guide
Design Immunohistochemistry for TOMM34
Plan TOMM34 paraffin IHC around cytoplasmic staining and high expression in spermatogonia (HPA tissue IHC). This guide uses the catalog antibody’s 2–5 μg/ml IHC range (datasheet A06775-1) and flags the medium consistency between staining and RNA data (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TOMM34 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); outer membrane pool (UniProt) | |
| Staining pattern | Cytoplasmic in spermatogonia, ovarian follicles and immune subsets (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06775-1) | |
| Positive control | Testis+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Medium staining–RNA consistency; verify independently (HPA tissue IHC) | |
| Regulation | No expression regulator reported (UniProt) | |
| Isoform / epitope | No isoforms; one 1–309 chain with no signal peptide (UniProt) |
Section 1
Recommended TOMM34 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A06775-1) is followed by published colon cancer and oral squamous cell carcinoma protocols (PMC9948080; PMC8262227).
| Sample | Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A06775-1) |
| Fixation | Image fixative and duration unreported (datasheet A06775-1); 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 A06775-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06775-1) |
| Primary antibody | Rabbit anti-TOMM34, 2-5 μg/ml (datasheet A06775-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06775-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06775-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TOMM34-positive staining in spermatogonia cells of testis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, highly abundant in spermatogonia in testis, ovarian follicles and subsets of immune cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A06775-1); the oral squamous cell carcinoma protocol used EDTA pH 9.0 (PMC8262227).
Section 2
What Is the Expected TOMM34 Staining Pattern?
TOMM34 should appear mainly cytoplasmic, with a possible mitochondrial component: UniProt places it in the cytoplasm and mitochondrial outer membrane and reports no transmembrane segment (UniProt Q15785 localization and topology). In tissue IHC, the strongest listed signal is in testicular spermatogonia; several glandular and immune cell populations also stain (HPA: tissue IHC). HPA rates the tissue profile Enhanced, with medium RNA–staining consistency and external verification pending (HPA: tissue reliability).
| Strong cytoplasmic staining in spermatogonia, with some other cell populations showing weaker staining. | This fits the listed tissue distribution: spermatogonia are High, while glandular cells in colon and adrenal gland and hematopoietic cells in bone marrow are Medium (HPA: tissue IHC). Compare intensity within identified cell types; a medium tissue need not match testis. |
| Signal appears cytoplasmic, sometimes with a more punctate component. | A mainly cytoplasmic result agrees with tissue IHC (HPA: tissue profile). A punctate component can be compatible with mitochondrial localization (UniProt Q15785 localization; HPA: ICC-IF), but chromogenic puncta alone do not identify mitochondria. |
| Staining is predominantly nuclear, with little cytoplasmic signal. | That compartment conflicts with the reported cytoplasm and mitochondrial outer membrane locations (UniProt Q15785 localization) and cytosolic ICC-IF pattern (HPA: subcellular). Treat it as suspect; review morphology, controls, and staining conditions before assigning TOMM34 expression. |
| Adipocytes or kidney glomerular cells stain strongly. | Those specific cell populations are listed as Not detected (HPA: adipose tissue; HPA: kidney). Check cell identity and no-primary detection controls for cross-reactivity or endogenous detection activity. The HPA calls apply to those cells, not every cell in either organ. |
| Spermatogonia show no convincing signal. | This conflicts with the High spermatogonial staining reported for testis (HPA: testis). Confirm that spermatogonia are present and assess the full IHC run with suitable controls before interpreting the sample as biologically negative. |
💡Expected TOMM34 appearanceA credible positive is High cytoplasmic signal in spermatogonia (HPA: testis), with possible mitochondrial-associated staining (UniProt Q15785 localization); strong nuclear-only signal or strong staining of listed Not detected cells is suspect (HPA: tissue IHC; HPA: subcellular).
| Cell type and tissue selection | Use spermatogonia in testis as the clearest listed positive reference (HPA: High in spermatogonia). Colon glandular cells or bone marrow hematopoietic cells offer Medium comparators (HPA: tissue IHC). Adipocytes and kidney glomerular cells are listed as Not detected in their respective tissues (HPA: tissue IHC). |
| Compartment and topology | The expected emphasis is cytoplasmic (HPA: tissue profile). TOMM34 is also assigned to the mitochondrial outer membrane, with no transmembrane segment (UniProt Q15785 localization and topology). This supports checking compartment, but does not make mitochondrial morphology alone a specificity test. |
| Strength of tissue evidence | HPA labels the tissue IHC profile Enhanced, while describing medium consistency with RNA and pending external verification (HPA: tissue reliability). Its two listed rabbit polyclonal antibodies each have Enhanced IHC validation (HPA: HPA018845; HPA056875). Use the cell-resolved pattern and controls when interpreting a new stain. |
| Fixation and antigen retrieval | Target-specific fixation sensitivity and retrieval requirements are unreported in the supplied UniProt and HPA records. As general IHC practice, assess retrieval using the chosen antibody's documented IHC-P conditions and positive tissue; do not infer a TOMM34-specific retrieval setting from expression or topology. |
| IF/ICC: what pattern is expected? | HPA reports mainly cytosolic signal, with additional vesicular and mitochondrial localization; cytosol and mitochondria are Supported, while vesicles are Approved (HPA: subcellular ICC-IF). That cell-based pattern can inform compartment interpretation, but does not establish an IHC-P dilution or retrieval condition. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in testicular spermatogonia | A known High population is absent from the stain, or the run failed (HPA: testis; general IHC practice). | Verify the cells on the section, then review the IHC-validated antibody's documented conditions, retrieval, detection reagents, and a concurrent positive control. Do not call the specimen negative until the run performs. |
| Uniform brown deposit across tissue and empty areas | Nonspecific background or detection chemistry can obscure cell boundaries (general chromogenic IHC practice). | Inspect a no-primary control; review blocking, washing, antibody concentration, and chromogen development. Judge TOMM34 only where signal remains cell associated and cytoplasmic (HPA: tissue profile). |
| Strong staining in adipocytes or glomerular cells | The result conflicts with their Not detected calls (HPA: adipose tissue; HPA: kidney); cross-reactivity or endogenous detection activity is possible. | Confirm the cell type and compare no-primary and appropriate detection controls. Reassess specificity before treating staining in those cells as TOMM34 expression. |
| Predominantly nuclear staining | The dominant compartment disagrees with cytoplasmic tissue staining and cytosolic ICC-IF localization (HPA: tissue profile; HPA: subcellular). | Check counterstain and cellular boundaries, review controls and staining conditions, and compare with a cytoplasmic positive reference. Do not score nuclear-only signal as the expected pattern. |
| Medium comparator looks much weaker than testis | That difference may match the reported levels: spermatogonia are High, while selected glandular and hematopoietic cells are Medium (HPA: tissue IHC). | Score the identified cell population against its own expected level and the same run's controls. Avoid requiring colon glandular cells or bone marrow hematopoietic cells to match spermatogonia. |
| Puncta dominate and diffuse cytoplasm is unclear | Mitochondrial localization is supported, but tissue IHC is described as cytoplasmic and ICC-IF as mainly cytosolic (UniProt Q15785 localization; HPA: tissue profile; HPA: subcellular). | Review morphology and controls for deposits or background. Interpret puncta cautiously in chromogenic IHC; use the separate IF/ICC guide for a cell-based compartment assessment. |
Experiment evidence
IHC & IF images showing TOMM34 staining
A06775-1 · IHCIHC analysis of TOMM34 using anti-TOMM34 antibody (A06775-1). TOMM34 was detected in a paraffin-embedded section of human prostate cancer ti
A06775-1 · IHCIHC analysis of TOMM34 using anti-TOMM34 antibody (A06775-1). TOMM34 was detected in a paraffin-embedded section of human cervical cancer ti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000025772-TOMM34/tissue/cerebral+cortex#imid_5018244
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000025772-TOMM34/tissue/colon#imid_5018170
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000025772-TOMM34/tissue/liver#imid_5018179
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000025772-TOMM34/tissue/kidney#imid_5018172
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA018845; green = TOMM34, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA018845; green = TOMM34, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TOMM34 IHC & IF
🧪Run testis first: spermatogonia should stain (HPA: High in spermatogonia). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the testis slide, neighboring cells without specific staining should remain at background, without assuming every other cell type is negative (HPA: spermatogonia High; UniProt Q15785: ubiquitous).
Positive control tissue: Testis (Spermatogonia cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TOMM34 in CACO-2, PC-3, U2OS, MCF-7, SiHa, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and rabbit IgG isotype controls matched to the primary antibody’s class and clonality where known (selected-SKU caption: rabbit primary; standard IHC practice). Confirm specificity with TOMM34 knockout material or a peptide block if an appropriate peptide is available, and quench endogenous peroxidase before HRP/DAB detection in testis (standard IHC practice; selected-SKU caption: HRP/DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A06775-1 tissue-IHC caption does not state a fixative (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required for TOMM34; neither frozen sections nor IF can be called easier from the supplied evidence (selected-SKU caption: EDTA retrieval; HPA: ICC-IF images). In testis, use morphology and counterstain to assign staining to spermatogonia within the crowded germ-cell layers (HPA: High in spermatogonia; standard IHC practice).
HPA tissue IHC evidence for TOMM34
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.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TOMM34 IHC Tips
Use the catalog antibody’s paraffin-section IHC conditions as a starting point, then judge TOMM34 staining by cell type, compartment and controls.
Which retrieval conditions should I start with for TOMM34 paraffin-section IHC?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A06775-1). The catalog antibody’s paraffin-section image used this retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody (datasheet A06775-1). Keep heating and cooling conditions identical across comparison sections, because uneven retrieval can resemble a biological difference (standard IHC practice). If staining is weak, vary heating duration on adjacent sections while keeping EDTA at pH 8.0; assess tissue integrity and background alongside signal (standard IHC practice). Record the final heating schedule, since the supplied caption does not specify one (datasheet A06775-1).
How should I handle fixation when TOMM34 staining varies between sections?
Target-specific fixation sensitivity is unknown: the catalog image identifies a paraffin section but does not state its fixative (datasheet A06775-1). For a controlled comparison, process matched specimens with the same fixative, fixation interval, section thickness and retrieval schedule (standard IHC practice). Compare a suspected weak section with a similarly processed positive control, such as spermatogonia, which show high staining in the tissue profile (HPA: High in spermatogonia). If staining differs, check morphology and processing records before changing antibody concentration (standard IHC practice). Do not infer a TOMM34-specific fixation effect from its reported localization or modified residues (UniProt Q15785; HPA tissue IHC).
What subcellular pattern should count as plausible TOMM34 staining?
Expect predominantly cytoplasmic staining in tissue sections (HPA tissue IHC: cytoplasmic expression in several tissues). TOMM34 is reported in the cytoplasm and at the mitochondrial outer membrane, with no transmembrane segment (UniProt Q15785 topology and subcellular location). A fine cytoplasmic or punctate pattern can therefore merit review, while a purely nuclear pattern warrants a specificity check (UniProt Q15785; HPA subcellular: cytosol, with additional mitochondrial and vesicular localization). Compare adjacent sections stained with and without primary antibody, then inspect signal at the same magnification and DAB development time (standard IHC practice). Chromogenic staining alone cannot establish mitochondrial colocalization (standard IHC practice).
Could an isoform or epitope difference explain inconsistent TOMM34 staining?
The supplied record lists 0 isoforms and a single 1–309 chain, so it provides no annotated isoform-specific explanation for discordant staining (UniProt Q15785). It also lists phosphoserines at positions 8, 160 and 186, but the antibody epitope is not supplied (UniProt Q15785; datasheet A06775-1). Do not attribute staining loss to phosphorylation without epitope information and direct experimental evidence (standard IHC interpretation). First compare retrieval, fixation records and staining-run controls on adjacent sections (standard IHC practice). If discrepancies persist, assess specificity with an independently validated antibody recognizing a documented different epitope (standard IHC practice).
How can IF help assess the IHC localization pattern?
Use IF as a separate validation experiment and multiplex TOMM34 with a marker for the cell population under study, such as spermatogonia in testis (HPA tissue IHC: High in spermatogonia). TOMM34 is mainly cytosolic, with additional mitochondrial and vesicular localization reported by subcellular imaging (HPA subcellular). Choose fluorophores after checking unstained tissue autofluorescence, and include single-label controls before interpreting overlap (standard IF practice). Because TOMM34 lacks a transmembrane segment and has a cytosolic location, use a permeabilisation condition that exposes intracellular epitopes, then check that cellular structure remains intact (UniProt Q15785 topology; standard IF practice). The paraffin-section caption supplies no IF fixation conditions (datasheet A06775-1).
What should I check when TOMM34 DAB staining looks diffuse or widespread?
Begin with a no-primary control and inspect whether brown signal persists in the same structures (standard chromogenic IHC practice). Block endogenous peroxidase and keep DAB development time consistent across sections; these are general chromogenic workflow steps, not TOMM34-specific evidence (standard IHC practice). The catalog image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary for 30 minutes at 37°C (datasheet A06775-1). If background remains high, compare shorter primary incubations or lower concentration on matched sections while retaining a positive control (standard IHC practice). Broad cytoplasmic staining alone is not proof of background because tissue expression is reported in several tissues (HPA tissue IHC).
How should I quantify TOMM34 IHC across different cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region of interest before scoring, then apply the same threshold and imaging settings to every section (standard IHC practice). For cell-associated DAB signal, report the percentage of positive cells and an H-score based on staining intensity and positive-cell fraction; state the intensity categories used (standard IHC practice). Normalize counts to the number of eligible cells, or report positive-cell density per mm² when cell enumeration is impractical (standard IHC practice). Score populations separately because high staining is reported in spermatogonia, while adipocytes are listed as not detected (HPA tissue IHC). Include run controls and exclude folds, necrosis and section edges consistently (standard IHC practice).
How can I distinguish convincing TOMM34 staining from an artefact?
A convincing result has intracellular cytoplasmic staining in an appropriate cell population and exceeds its matched no-primary control (HPA tissue IHC; standard IHC practice). High staining in spermatogonia offers a positive reference, while HPA lists adipocytes as not detected; these are reference patterns rather than guarantees for every specimen (HPA tissue IHC). Treat exclusively nuclear signal cautiously because the supplied localization evidence favors cytosol, with additional mitochondrial and vesicular localization (UniProt Q15785; HPA subcellular). Review edge-restricted staining, necrotic areas and persistent signal after primary omission for processing artefact or endogenous enzyme activity (standard IHC practice). The tissue IHC profile has medium staining–RNA consistency and awaits external verification (HPA tissue IHC: Enhanced reliability description).
Boster reagents
Best TOMM34 / Mitochondrial import receptor subunit TOM34 IHC Antibodies
A06775-1 has real IHC images from paraffin-embedded human prostate, cervical, liver and lung cancer sections (catalog image captions). The antibody lists human and rat IHC use; no IF data are supplied (catalog applications).
Real IHC data
Anti-TOMM34 Antibody ®
Cat # A06775-1
A06775-1 is the only SKU that will render, with IHC images from paraffin-embedded human prostate, cervical, liver and lung cancer sections (catalog image captions). Its application data list IHC for human and rat samples; the broader reactivity list also includes mouse, but no mouse IHC image or IF validation is supplied (catalog application and reactivity data).
Which to pick: For paraffin-section tissue IHC, choose A06775-1: its own images document heat retrieval in EDTA at pH 8.0 and primary antibody at 2 μg/ml (catalog image captions). A06775-1 is the only listed cross-species option, with human, mouse and rat reactivity, though IHC is listed for human and rat only (catalog application and reactivity data). No SKU is listed for IF/ICC (catalog applications); the paraffin-section captions do not report the fixative (catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q15785 (TOM34_HUMAN, Mitochondrial import receptor subunit TOM34).
- Human Protein Atlas. TOMM34 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. TOMM34 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to vesicles and mitochondria..
- Human Protein Atlas. TOMM34 antibody validation summary (2 antibodies).
- Tomm34 is commonly expressed in epithelial ovarian cancer and associates with tumour type and high FIGO stage. Journal of ovarian research 2019 — PMC6436220.
- TOMM34 serves as a candidate therapeutic target associated with immune cell infiltration in colon cancer. Frontiers in oncology 2023 — PMC9948080.
- High Expression of Tomm34 and Its Correlations With Clinicopathology in Oral Squamous Cell Carcinoma. Pathology oncology research : POR 2021 — PMC8262227.
- Disrupting metformin adaptation of liver cancer cells by targeting the TOMM34/ATP5B axis. EMBO molecular medicine 2022 — PMC9728056.
- PubMed PMID:9324309 — UniProt-cited evidence.
- PubMed PMID:16820880 — UniProt-cited evidence.
- PubMed PMID:11780052 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TOMM34 (Mitochondrial import receptor subunit TOM34). Expected localisation Cytoplasmic staining (HPA tissue IHC); outer membrane pool (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06775-1). Validated IHC antibody: A06775-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Spermatogonia cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A06775-1); 2. Fixation: Image fixative and duration unreported (datasheet A06775-1); 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 A06775-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06775-1); 8. Primary antibody: Rabbit anti-TOMM34, 2-5 μg/ml (datasheet A06775-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06775-1). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Medium staining–RNA consistency; verify independently (HPA tissue IHC); Isoform / epitope: No isoforms; one 1–309 chain with no signal peptide (UniProt); Regulation: No expression regulator reported (UniProt). Evidence: UniProt Q15785, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to vesicles…, Datasheet A06775-1, PMC9948080, PMC8262227. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TOMM34/immunohistochemistry-ihc-guide.