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TIMM17A / Mitochondrial import inner membrane translocase subunit Tim17-A · IHC design guide
Design Immunohistochemistry for TIMM17A
For paraffin-section TIMM17A IHC, start with 0.5–1 μg/mL of the catalog antibody (datasheet A12168-1). Assess cytoplasmic staining using high-staining cardiomyocytes as a reference, while accounting for the medium consistency between antibody staining and RNA data (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TIMM17A 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 tissue staining (HPA tissue IHC); mitochondrial inner membrane (UniProt) | |
| Staining pattern | Cytoplasmic staining; high in cardiomyocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A12168-1) | |
| Positive control | Esophagus+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 consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has medium consistency with RNA data (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms or cleavage annotated; 3 transmembrane segments (UniProt) |
Section 1
Recommended TIMM17A IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published TIMM17A IHC protocol using rat tissue slides (PMC6257833: Immunohistochemical detection).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A12168-1) |
| Fixation | Image fixative and duration unreported (datasheet A12168-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A12168-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A12168-1) |
| Primary antibody | Rabbit anti-TIMM17A, 0.5-1μg/ml (datasheet A12168-1) |
| Primary incubation | Overnight at 4 °C (datasheet A12168-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A12168-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TIMM17A-positive staining in squamous epithelial cells of esophagus (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval for the catalog antibody (datasheet A12168-1). The published rat protocol does not specify retrieval (PMC6257833: Immunohistochemical detection).
Section 2
What Is the Expected TIMM17A Staining Pattern?
TIMM17A resides in the mitochondrial inner membrane and has 3 transmembrane segments (UniProt Q99595 topology). In paraffin section IHC, expect cytoplasmic staining consistent with that location; HPA describes general cytoplasmic expression and rates its tissue IHC evidence Approved, with medium consistency between staining and RNA data (HPA tissue IHC). High staining is reported in squamous epithelial cells, cardiomyocytes, and selected glandular cells (HPA tissue IHC).
| Cytoplasmic stain in esophageal or oral squamous epithelium, cardiomyocytes, or rectal or thyroid glandular cells. | These cells have High HPA tissue IHC staining. A cytoplasmic pattern fits mitochondrial localization; chromogenic IHC need not resolve individual mitochondria (HPA tissue IHC; UniProt Q99595 localization; standard IHC practice). |
| Predominantly nuclear or extracellular stain, with little cytoplasmic signal in an HPA High cell population. | This poorly fits the mitochondrial inner membrane assignment (UniProt Q99595 localization). Review controls before calling it specific; HPA also lists nucleoplasm as an uncertain IF location, so nuclear signal alone cannot settle specificity (HPA subcellular ICC-IF; standard IHC practice). |
| Strong stain in a cell type that lacks an established positive pattern in this payload. | Possible cross-reactivity or endogenous detection activity warrants checking (standard IHC practice). Absence from HPA's positive list is not evidence that the cell type must be negative; HPA reports low tissue specificity (HPA tissue IHC). |
| Widespread, even stain over cells and surrounding tissue, obscuring cell boundaries. | Diffuse background limits interpretation of the expected cytoplasmic pattern (HPA tissue IHC; standard IHC practice). Assess a no-primary control and the detection and blocking steps before assigning TIMM17A expression (standard IHC practice). |
| No stain in a well-preserved HPA High population on the same run. | A technical failure becomes plausible when a documented positive is absent, though one negative section cannot establish its cause (HPA tissue IHC; standard IHC practice). Check controls, retrieval, antibody dilution, and detection (standard IHC practice). |
💡Expected TIMM17A appearanceCall positive when staining is chiefly cytoplasmic in an HPA High cell population, such as cardiomyocytes or rectal glandular cells; an isolated strong nuclear or tissue-wide diffuse signal needs control-based review (HPA tissue IHC; UniProt Q99595 localization; standard IHC practice).
| Compartment and membrane topology | TIMM17A is an inner mitochondrial membrane protein with 3 |
| Tissue and cell choice | HPA reports High staining in esophageal and oral squamous epithelium, cardiomyocytes, and rectal and thyroid glandular cells; pancreatic exocrine cells and skeletal myocytes are Low (HPA tissue IHC). Compare like cell types when judging intensity (standard IHC practice). |
| Strength of IHC evidence | HPA calls tissue IHC Approved, with medium staining-to-RNA consistency; antibody HPA010083 is IHC Approved (HPA tissue IHC; HPA antibodies). These labels support use of the observed pattern but do not prove that every stained cell is specific (standard IHC practice). |
| Processing and modification record | The UniProt record lists one 171-aa chain, no signal peptide or propeptide, and no annotated glycosylation or modified residues (UniProt Q99595). It supplies no basis here for predicting a distinct processed staining compartment (UniProt Q99595). |
| Q: What does IF/ICC add? | A: HPA reports enhanced mitochondrial and uncertain nucleoplasmic localization; its IF images include A-549, SiHa, and U2OS (HPA subcellular ICC-IF). This context helps assess compartment claims, while IF/ICC methods belong to the separate guide (HPA subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in esophageal squamous cells or cardiomyocytes. | Both are HPA High populations, so absent stain raises concern about the IHC run or specimen quality (HPA tissue IHC; standard IHC practice). | Confirm tissue preservation and a positive control; review retrieval, antibody dilution, and detection against the validated IHC procedure (standard IHC practice). |
| Weak signal in pancreatic exocrine cells or skeletal myocytes. | Both are listed Low in HPA tissue IHC; weak staining there can fit the reported pattern (HPA tissue IHC). | Judge run performance in an HPA High population on the same run before changing conditions (HPA tissue IHC; standard IHC practice). |
| Nuclear stain dominates an HPA High cell population. | Mitochondria are the enhanced IF location, while nucleoplasm is uncertain; nuclear dominance has ambiguous specificity (HPA subcellular ICC-IF; UniProt Q99595 localization). | Compare cytoplasmic staining and a no-primary control, then review the IHC-validated antibody's pattern; do not score nuclear stain alone as a confirmed positive (HPA tissue IHC; standard IHC practice). |
| Broad stain persists in a no-primary control. | Signal without primary antibody points toward detection-system or endogenous activity rather than TIMM17A binding (standard IHC practice). | Review blocking and the chromogenic detection controls; interpret the primary-stained slide only after control background is resolved (standard IHC practice). |
| Unexpected cells stain strongly while expected cells do not. | Cross-reactivity or uneven assay performance is possible; HPA's low tissue specificity prevents treating every unlisted cell as a known negative (HPA tissue IHC; standard IHC practice). | Compare cell identity, compartment, and matched controls with an HPA High reference; avoid assigning specificity from intensity alone (HPA tissue IHC; standard IHC practice). |
| Cytoplasmic stain is too diffuse to score individual cells. | Excess background can obscure the general cytoplasmic profile reported by HPA (HPA tissue IHC; standard IHC practice). | Check no-primary background, blocking, antibody dilution, and detection conditions before scoring cell-level intensity (standard IHC practice). |
Experiment evidence
IHC & IF images showing TIMM17A staining
A12168-1 · IHCIHC analysis of TIMM17A using anti-TIMM17A antibody (A12168-1). TIMM17A was detected in paraffin-embedded section of human liver cancer tiss
A12168-1 · IHCIHC analysis of TIMM17A using anti-TIMM17A antibody (A12168-1). TIMM17A was detected in paraffin-embedded section of human lung cancer tissu
A12168-1 · IFIF analysis of TIMM17A using anti-TIMM17A antibody (A12168-1). TIMM17A was detected in immunocytochemical section of HELA cell. Enzyme antig
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000134375-TIMM17A/tissue/cerebral+cortex#imid_7479569
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000134375-TIMM17A/tissue/colon#imid_7477543
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000134375-TIMM17A/tissue/liver#imid_7477536
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000134375-TIMM17A/tissue/kidney#imid_7477545
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA010083; green = TIMM17A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA010083; green = TIMM17A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TIMM17A IHC & IF
🧪Run heart muscle first; cardiomyocytes should stain (HPA: High in cardiomyocytes). HPA detects TIMM17A in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and expect erythrocytes within the positive section to lack mitochondrial staining (HPA: no negative tissue; standard IHC practice).
Positive control tissue: Esophagus (Squamous epithelial cells, HPA High)
Negative control tissue: None in HPA: TIMM17A 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 TIMM17A in A-549, SiHa, U2OS, with annotated localisation: Nucleoplasm (uncertain), Mitochondria (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a matched rabbit IgG isotype control, and TIMM17A-knockout material as a biological negative if available (caption A12168-1: rabbit primary; standard IHC practice). Block endogenous peroxidase and, if using the caption’s biotin-based detection, assess endogenous biotin in heart muscle (caption A12168-1: biotinylated secondary and SABC–DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state its fixative (caption A12168-1). The caption uses citrate retrieval at pH 6 for 20 minutes, but does not establish whether retrieval is required or optimal (caption A12168-1). Frozen-section versus IF ease is unreported; heart muscle can contribute endogenous biotin background with biotin-based detection (caption A12168-1: SABC–DAB; standard IHC practice).
HPA tissue IHC evidence for TIMM17A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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 |
|---|---|---|---|---|
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Rectum | Glandular cells | High | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: TIMM17A is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced TIMM17A IHC Tips
Troubleshooting guidance for TIMM17A chromogenic IHC in paraffin sections, with one note on adapting the experiment to IF.
What retrieval conditions should I try when TIMM17A staining is weak?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes in paraffin sections (datasheet A12168-1). Keep the catalog antibody concentration at 1 µg/mL and its overnight 4°C incubation initially, so retrieval is the variable under test (datasheet A12168-1; standard IHC practice). If staining remains weak, compare longer heating or an alternative retrieval buffer on matched sections while checking tissue integrity and background (standard IHC practice). TIMM17A has 3 predicted transmembrane segments, so accessibility may depend on the antibody’s epitope, which is not specified here (UniProt Q99595 topology; datasheet A12168-1).
How should I troubleshoot variable staining between differently fixed sections?
The selected TIMM17A tissue image describes a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A12168-1). Record each specimen’s fixative, fixation duration, processing conditions and section age before comparing staining across batches (standard IHC practice). Run matched sections through the same pH 6, 20-minute citrate retrieval and 1 µg/mL primary antibody incubation to isolate processing differences (datasheet A12168-1; standard IHC practice). Compare preservation, staining distribution and negative controls before changing retrieval, since tissue damage or uneven processing can distort chromogenic signal (standard IHC practice).
Should I score diffuse cytoplasmic or nuclear TIMM17A staining as positive?
Prioritise a cytoplasmic, mitochondria-compatible pattern: TIMM17A is assigned to the mitochondrial inner membrane, while tissue IHC is described as generally cytoplasmic (UniProt Q99595 subcellular; HPA tissue IHC). Chromogenic IHC may not resolve individual mitochondria, so assess the pattern within intact cells alongside a matched negative control (standard IHC practice). Treat predominantly nuclear staining cautiously because the HPA mitochondrial IF location is enhanced, whereas its nucleoplasmic location is uncertain (HPA subcellular). Compare the same cell population across sections, since high HPA tissue IHC staining is reported in cardiomyocytes and several epithelial or glandular cell populations (HPA tissue IHC).
Could an isoform or hidden membrane epitope explain discordant staining?
The supplied UniProt record lists 0 isoforms and a 171-amino-acid chain, so it gives no annotated isoform-specific staining pattern to expect (UniProt Q99595 record). Its 3 transmembrane segments span residues 17–37, 63–77 and 113–133, making epitope accessibility a practical question for section staining (UniProt Q99595 topology; standard IHC practice). The antibody epitope is not specified here, so do not assign a membrane-facing side or infer an epitope-specific retrieval requirement (datasheet A12168-1). Instead, compare serial sections under the documented pH 6 retrieval and controlled retrieval adjustments, assessing signal and tissue preservation together (datasheet A12168-1; standard IHC practice).
How can I assess TIMM17A by multiplex IF alongside cell identity?
For IF, pair TIMM17A with a marker identifying the cell population being assessed, such as a cardiomyocyte marker in heart muscle, where HPA reports high staining (HPA tissue IHC; standard IF practice). Choose fluorophores after checking tissue autofluorescence and include single-stain controls to distinguish bleed-through from colocalisation (standard IF practice). TIMM17A resides in the inner mitochondrial membrane and has 3 predicted transmembrane segments, but the antibody epitope’s membrane-facing side is unspecified (UniProt Q99595 topology; datasheet A12168-1). Optimise permeabilisation for access to the actual epitope and confirm the expected mitochondrial pattern; the paraffin-section IHC conditions do not establish an IF protocol (standard IF practice; datasheet A12168-1; HPA subcellular).
What should I change when the chromogenic signal obscures TIMM17A staining?
Begin with a section lacking primary antibody, then inspect pigment, tissue edges and damaged areas before adjusting antibody concentration (standard IHC practice). The selected IHC image used 10% goat serum, 1 µg/mL primary antibody, biotinylated secondary antibody and DAB development (datasheet A12168-1). If background persists, titrate primary antibody, shorten chromogen development and check blocking and washes while retaining a matched positive control (standard IHC practice). Include appropriate endogenous peroxidase and biotin controls for a peroxidase-based avidin–biotin workflow, since those signals can mimic cellular DAB staining (standard IHC practice; datasheet A12168-1).
How should I quantify TIMM17A across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then apply the same segmentation and intensity thresholds to every section (standard IHC practice). For cellular staining, report the percentage of positive cells and an H-score using intensity categories; document the scoring scale and observer agreement (standard IHC practice). If counting discrete positive cells, report density per mm² of viable tissue and exclude folds, necrosis and section edges by a predefined rule (standard IHC practice). Normalise comparisons to the relevant viable cell population or analysed tissue area, and keep retrieval, antibody incubation and DAB development consistent across batches (standard IHC practice; datasheet A12168-1).
How do I distinguish credible TIMM17A staining from an artefact?
A credible result shows reproducible cellular staining compatible with mitochondrial localisation, interpreted at the resolution of chromogenic tissue IHC (UniProt Q99595 subcellular; HPA tissue IHC; standard IHC practice). Check which cells carry signal: HPA reports high staining in cardiomyocytes and selected squamous epithelial or glandular cells, with medium consistency between staining and RNA data (HPA tissue IHC). Predominantly nuclear signal warrants caution because HPA calls the nucleoplasmic IF location uncertain, while its mitochondrial location is enhanced (HPA subcellular). Reject edge-only, necrotic or primary-omission signal as evidence of TIMM17A, and check endogenous peroxidase before attributing DAB deposits to the target (standard IHC practice).
Boster reagents
Best TIMM17A / Mitochondrial import inner membrane translocase subunit Tim17-A IHC Antibodies
A12168-1 has real paraffin-section IHC images from human cancer tissues and rat lung, plus IF/ICC images from HeLa and A549 cells (catalog image captions).
Real IHC data
Anti-TIMM17A Antibody ®
Cat # A12168-1
A12168-1 is listed for IHC and IF/ICC in human, mouse, and rat (catalog applications and reactivity). Its IHC images show human liver, lung, and mammary cancer sections and rat lung; its IF images show HeLa and A549 cells (catalog image captions).
Which to pick: For chromogenic IHC on paraffin sections, choose A12168-1: its own caption documents citrate retrieval at pH 6 and primary antibody at 1 μg/ml; the fixative and clone are unreported (catalog IHC caption; catalog clone field). For IF/ICC, A12168-1 has images from HeLa and A549 cells (catalog IF captions). For cross-species work, A12168-1 lists human, mouse, and rat reactivity; rat tissue IHC is pictured, while mouse tissue IHC is unpictured (catalog reactivity; catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q99595 (TI17A_HUMAN, Mitochondrial import inner membrane translocase subunit Tim17-A).
- Human Protein Atlas. TIMM17A tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TIMM17A subcellular location (ICC-IF): Localized to the nucleoplasm and mitochondria..
- Human Protein Atlas. TIMM17A antibody validation summary (1 antibodies).
- 'Governor vessel-unblocking and mind-regulating' acupuncture therapy ameliorates cognitive dysfunction in a rat model of middle cerebral artery occlusion. International journal of molecular medicine 2019 — PMC6257833.
- PubMed PMID:8893850 — UniProt-cited evidence.
- PubMed PMID:10339406 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TIMM17A (Mitochondrial import inner membrane translocase subunit Tim17-A). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); mitochondrial inner membrane (UniProt). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A12168-1). Validated IHC antibody: A12168-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Esophagus (Squamous epithelial 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 human liver cancer tissue; fixative not specified (datasheet A12168-1); 2. Fixation: Image fixative and duration unreported (datasheet A12168-1); 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: Citrate pH 6 , 20 min (datasheet A12168-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A12168-1); 8. Primary antibody: Rabbit anti-TIMM17A, 0.5-1μg/ml (datasheet A12168-1); 9. Primary incubation: Overnight at 4 °C (datasheet A12168-1). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Antibody staining has medium consistency with RNA data (HPA tissue IHC); Isoform / epitope: No isoforms or cleavage annotated; 3 transmembrane segments (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q99595, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm and mitochondria., Datasheet A12168-1, PMC6257833. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TIMM17A/immunohistochemistry-ihc-guide.