This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
CLPB / Mitochondrial disaggregase · IHC design guide
Design Immunohistochemistry for CLPB
Plan CLPB paraffin IHC around granular cytoplasmic staining in kidney tubules and seminal vesicle glands (HPA tissue IHC). This guide covers fixation consistency, an antibody range of 0.5–1 μg/ml (datasheet A02912-2), and interpretation alongside mitochondrial intermembrane space localization (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CLPB 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 | Granular cytoplasm (HPA tissue IHC) | |
| Staining pattern | Tubular and glandular cells: granular cytoplasm (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02912-2) | |
| Positive control | Kidney+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 sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA expression show very low consistency (HPA tissue IHC) | |
| Regulation | Low expression in granulocytes (UniProt) | |
| Isoform / epitope | 5 isoforms; N-terminal epitopes may be lost after cleavage (UniProt) |
Section 1
Recommended CLPB IHC & IF Protocols
The catalog antibody protocol uses EDTA retrieval (datasheet A02912-2). The published heart tissue IHC protocol below uses citrate retrieval (PMC11641810).
| Sample | Paraffin-embedded human mammary cancer tissue; fixative not specified (datasheet A02912-2) |
| Fixation | Image fixative and duration unreported (datasheet A02912-2); 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 A02912-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02912-2) |
| Primary antibody | Rabbit anti-CLPB, 0.5-1μg/ml (datasheet A02912-2) |
| Primary incubation | Overnight at 4 °C (datasheet A02912-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02912-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CLPB-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A02912-2); compare the published citrate method if staining needs optimization (PMC11641810).
Section 2
What Is the Expected CLPB Staining Pattern?
CLPB is a mitochondrial intermembrane space protein without a transmembrane segment (UniProt Q9H078 topology). In paraffin sections, expect granular cytoplasmic staining, especially in kidney tubular cells and seminal vesicle glandular cells (HPA: general granular cytoplasmic profile; High in both cell types). HPA rates tissue IHC reliability Enhanced, while reporting very low agreement with RNA expression and pending external verification (HPA: reliability description).
| Granular cytoplasmic stain in kidney tubular cells or seminal vesicle glandular cells. | This matches the reported High staining in those cells (HPA: tissue IHC). Assess the granules within cell boundaries; routine chromogenic IHC cannot resolve the mitochondrial intermembrane space (UniProt Q9H078 localisation; standard IHC practice). |
| Predominantly nuclear or sharply cell-surface staining, with little cytoplasmic granularity. | The compartment conflicts with CLPB's mitochondrial location and HPA's cytoplasmic pattern (UniProt Q9H078; HPA: tissue profile). Treat it as suspect and check antibody specificity and section artefacts (standard IHC practice). |
| Strong signal chiefly in cells other than the reported positive cell population. | For that tissue, consider cross-reactivity or endogenous chromogenic activity (HPA: cell-level tissue IHC; standard IHC practice). Do not classify every unlisted cell as negative: HPA supplies no confirmed negative tissues (HPA: tissue IHC). |
| Uniform diffuse color across cells, lumina, and surrounding tissue. | This does not resemble the reported granular cytoplasmic profile (HPA: tissue IHC). Background from detection chemistry or insufficient blocking may obscure interpretable cellular signal (standard IHC practice). |
| No cellular signal in kidney tubular cells or seminal vesicle glandular cells. | These are useful positive benchmarks because both stain High in HPA tissue IHC (HPA: tissue IHC). A blank section calls for a workflow and antibody check before interpreting the specimen as CLPB absent (standard IHC practice). |
💡Expected CLPB appearanceCall a positive result when chromogenic signal is granular and cytoplasmic in kidney tubular or seminal vesicle glandular cells, where HPA reports High staining; dominant nuclear or cell-surface color is suspect (HPA: tissue IHC; UniProt Q9H078 localisation).
| Reported cell populations | HPA reports Medium staining in several glandular and neuronal populations, alongside High kidney tubular and seminal vesicle glandular staining (HPA: tissue IHC). Match intensity to the cell type rather than assigning one intensity to every tissue. |
| Protein processing and epitope coverage | UniProt lists CLPB chains spanning residues 37–707 and 127–707 (UniProt Q9H078 processing). Epitope position is not supplied, so antibody recognition of both forms cannot be inferred. |
| Isoforms | UniProt lists five CLPB isoforms (UniProt Q9H078 isoforms). The payload gives no antibody epitope or isoform-specific tissue pattern; avoid attributing staining differences to a particular isoform. |
| Antibody evidence and retrieval | Two listed antibodies are IHC Enhanced and one is Approved (HPA: antibody validation). Target-specific fixation sensitivity and retrieval conditions are unreported here; optimize retrieval against controls as general IHC practice. |
| IF/ICC: should the same pattern be expected? | CLPB's mitochondrial location suggests cytoplasmic organelle-associated signal (UniProt Q9H078 localisation), but HPA supplies no ICC-IF images or verified IF location (HPA: subcellular record). Assess IF/ICC separately; an IHC pattern does not validate an IF assay. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive benchmark is blank. | The run may have lost detectable signal; HPA reports High staining in kidney tubular and seminal vesicle glandular cells (HPA: tissue IHC). | Check control-section processing, primary antibody use, retrieval, and chromogenic detection; adjust conditions against the positive benchmark (standard IHC practice). |
| Granules are weak but broad haze is strong. | Background can conceal the granular cytoplasmic profile reported by HPA (HPA: tissue profile; standard IHC practice). | Review blocking, washes, primary antibody concentration, and detection exposure with matched controls (standard IHC practice). |
| Strong nuclear or membrane-edge color dominates. | That distribution conflicts with mitochondrial CLPB and HPA's cytoplasmic profile (UniProt Q9H078; HPA: tissue IHC). | Check section artefacts and detection controls; compare with an independently validated antibody where available (HPA: antibody validation; standard IHC practice). |
| Color appears without primary antibody. | Endogenous detection activity or nonspecific detection can produce chromogenic color (standard IHC practice). | Use the matched no-primary control to locate background, then review endogenous-activity quenching and detection reagents (standard IHC practice). |
| An unlisted tissue or cell type stains strongly. | HPA lists selected cell-level results and no confirmed negative tissues, so specificity cannot be decided from an unlisted site alone (HPA: tissue IHC). | Compare compartment and granularity with the reported pattern, and use a second antibody or orthogonal control if the finding matters (HPA: tissue profile; standard IHC practice). |
| Intensity differs from RNA abundance. | HPA explicitly reports very low consistency between antibody staining and RNA expression, despite its Enhanced tissue IHC rating (HPA: reliability description). | Report the observed cell-level stain and control results separately from RNA data; avoid treating their agreement as proof of specificity (HPA: reliability description; standard IHC practice). |
Experiment evidence
IHC & IF images showing CLPB staining
A02912-2 · IHCIHC analysis of CLPB using anti-CLPB antibody (A02912-2). CLPB was detected in paraffin-embedded section of human mammary cancer tissue. Hea
A02912-2 · IHCIHC analysis of CLPB using anti-CLPB antibody (A02912-2). CLPB was detected in paraffin-embedded section of human rectal cancer tissue. Heat
A02912-2 · IFIF analysis of CLPB using anti-CLPB antibody (A02912-2). CLPB was detected in immunocytochemical section of A549 cells. Enzyme antigen retri
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000162129-CLPB/tissue/cerebral+cortex#imid_18093167
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000162129-CLPB/tissue/adrenal+gland#imid_18093143
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000162129-CLPB/tissue/colon#imid_18093093
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000162129-CLPB/tissue/liver#imid_18093094
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CLPB IHC & IF
🧪Run kidney first; cells in tubules should stain (HPA: High in cells in tubules). HPA detects CLPB in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat cells outside the stained tubules as internal background references rather than proven CLPB-negative cells (HPA: detected in all 45 tissues).
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: None in HPA: CLPB 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 carries no ICC-IF cell line for CLPB; derive a cell-line control from the positive tissue's cell type (Cells in tubules) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control, a host- and clonality-matched rabbit isotype control, and a CLPB-knockout biological control where available (caption: rabbit anti-CLPB primary; standard IHC practice). In kidney, check endogenous peroxidase and biotin background before interpreting SABC/DAB staining (caption: SABC/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the A02912-2 paraffin-section caption does not state a fixative (caption: fixative unreported). The reported IHC procedure uses heat-mediated EDTA retrieval at pH 8.0, but the evidence does not establish that retrieval is essential (caption: EDTA retrieval). Frozen sections and IF/ICC cannot be judged easier from the supplied evidence; kidney endogenous biotin may complicate the reported SABC detection (HPA: no ICC-IF images; caption: SABC detection; standard IHC practice).
HPA tissue IHC evidence for CLPB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Very low consistency between antibody staining and RNA expression data. External characterization data supports antibody staining. 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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: CLPB is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced CLPB IHC Tips
Troubleshoot CLPB staining in paraffin sections by checking retrieval, compartment, cell type and controls before comparing chromogenic IHC scores.
What retrieval should I use if CLPB staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for CLPB paraffin-section IHC (datasheet A02912-2). The selected image used this retrieval before incubation with 1 μg/mL antibody overnight at 4°C, so reproduce those conditions when assessing weak staining (datasheet A02912-2). If staining remains weak, compare a shorter and longer heating interval on matched sections while keeping buffer, detection and imaging conditions constant (standard IHC practice). Inspect tissue preservation and background alongside signal, because stronger staining after harsher heating alone does not establish that the signal is CLPB-specific (standard IHC practice).
How should I assess whether fixation is causing weak CLPB IHC staining?
The selected CLPB image documents a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A02912-2). Record the actual fixative and fixation duration for each specimen, then compare matched sections processed with the same EDTA retrieval at pH 8.0 (datasheet A02912-2; standard IHC practice). Include a positive-control section in each run and assess morphology as well as staining intensity before changing retrieval or antibody concentration (standard IHC practice). Do not attribute a fixation effect to CLPB’s mitochondrial location, processing or acetylation without a controlled comparison (UniProt Q9H078; standard IHC practice).
Which staining pattern is plausible for CLPB in chromogenic tissue IHC?
Look for cytoplasmic granular staining rather than a uniform nuclear signal: tissue IHC describes CLPB as generally cytoplasmic with a granular pattern (HPA: tissue IHC profile). This is compatible with its annotated location in the mitochondrial intermembrane space, although DAB staining alone cannot resolve that compartment (UniProt Q9H078; standard IHC practice). For a tissue control, kidney tubule cells show high staining, while breast glandular cells show medium staining in the cited atlas (HPA: tissue IHC). Score the relevant cells separately from neighboring tissue and compare their pattern with background and control sections processed in the same run (standard IHC practice).
Could CLPB processing or isoforms change what the antibody detects in sections?
CLPB has five annotated isoforms and reported protein chains spanning residues 37–707 and 127–707 (UniProt Q9H078). An antibody to an epitope absent from a processed chain could stain differently from one recognizing a retained region, but the supplied evidence does not map this antibody’s epitope (UniProt Q9H078; datasheet A02912-2). Check the immunogen or epitope documentation before assigning an IHC pattern to a particular isoform or cleavage state (standard IHC practice). If such documentation is unavailable, report staining as CLPB immunoreactivity and use an independently validated antibody or orthogonal assay to investigate discordant cases (standard IHC practice).
How can IF help check a questionable CLPB IHC pattern?
On a matched section, multiplex CLPB with a validated marker for the expected cell type, such as a tubular epithelial marker when evaluating kidney tubules (HPA: high in kidney tubule cells; standard IF practice). Choose spectrally separated fluorophores after inspecting unstained tissue autofluorescence, and use single-label controls to assess channel bleed-through (standard IF practice). CLPB is annotated in the mitochondrial intermembrane space, so titrate permeabilisation to allow antibody access across mitochondrial membranes while retaining organelle morphology; the antibody’s epitope is not mapped here (UniProt Q9H078; standard IF practice). Treat concordant cellular staining as supporting evidence, while checking IF and IHC controls independently (standard IF/IHC practice).
How do I reduce diffuse or granular background without losing CLPB signal?
First compare the stained section with a no-primary control and inspect whether background follows tissue edges, damaged areas or endogenous pigment (standard IHC practice). The selected CLPB image used 10% goat serum blocking, 1 μg/mL primary antibody overnight at 4°C, and a biotin-based DAB detection system (datasheet A02912-2). Include a peroxidase block for chromogenic detection and assess endogenous biotin interference when using a biotin-based system (standard IHC practice). If background persists, titrate the primary antibody on matched sections and judge signal against cellular localisation and control staining, rather than intensity alone (standard IHC practice).
How should I quantify CLPB staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring; kidney tubule cells and breast glandular cells have different reported staining levels (HPA: high in kidney tubules; medium in breast glandular cells). For chromogenic sections, record the percentage of positive target cells and an H-score using intensity categories 0–3, or measure positive-cell density per mm² when cell counts are the outcome (standard IHC practice). Normalize to the number of eligible cells or evaluable tissue area, and exclude folds, necrosis and poorly preserved regions by predefined rules (standard IHC practice). Apply identical retrieval, detection, threshold and scoring rules across comparison groups (standard IHC practice).
What separates credible CLPB positivity from an IHC artefact?
Credible CLPB staining should show a cellular, granular cytoplasmic pattern consistent with tissue IHC and its annotated mitochondrial location (HPA: tissue IHC profile; UniProt Q9H078). Assess the expected cell population explicitly: high staining is reported in kidney tubule cells, whereas low staining is reported in hippocampal glial cells (HPA: tissue IHC). Treat isolated nuclear colour, edge-heavy signal, necrotic deposits or staining reproduced in a no-primary control as reasons to investigate artefact (standard IHC practice). Check endogenous peroxidase and, for the selected biotin-based DAB workflow, detection-system background before interpreting discordant staining as CLPB expression (datasheet A02912-2; standard IHC practice).
Boster reagents
Best CLPB / Mitochondrial disaggregase IHC Antibodies
A02912-2 has IHC data from human paraffin sections and IF/ICC data from A549 cells (catalog image captions); catalog reactivity includes human, monkey, mouse and rat (catalog reactivity).
Real IHC data
Anti-CLPB Antibody ®
Cat # A02912-2
A02912-2 will render with an IHC figure from a human mammary cancer paraffin section; its other IHC caption describes a human rectal cancer paraffin section (catalog IHC captions). Its IF caption documents A549 cells stained at 4 μg/ml (catalog IF caption).
Which to pick: Choose A02912-2 for human paraffin-section IHC: its own caption documents EDTA retrieval at pH 8.0 and primary antibody at 1 μg/ml; the fixative is unreported (catalog IHC caption). Choose the same SKU for IF/ICC in A549 cells, supported by its IF caption (catalog IF caption). For other species, the catalog lists monkey, mouse and rat reactivity, but the supplied IHC and IF captions document human samples only (catalog reactivity; IHC/IF captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H078 (CLPB_HUMAN, Mitochondrial disaggregase).
- Human Protein Atlas. CLPB tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CLPB subcellular location (ICC-IF): Highest expression in SK-MEL-30: 54.5 nTPM.
- Human Protein Atlas. CLPB antibody validation summary (3 antibodies).
- Neuronal mitochondrial disaggregase CLPB ameliorates Huntington's disease pathology in mice. Theranostics 2026 — PMC12712926.
- Exploring Potential Diagnostic Biomarkers for Mechanical Asphyxia in the Heart Based on Proteomics Technology. International journal of molecular sciences 2024 — PMC11641810.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16554811 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CLPB (Mitochondrial disaggregase). Expected localisation Granular cytoplasm (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02912-2). Validated IHC antibody: A02912-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); 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 mammary cancer tissue; fixative not specified (datasheet A02912-2); 2. Fixation: Image fixative and duration unreported (datasheet A02912-2); 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 A02912-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02912-2); 8. Primary antibody: Rabbit anti-CLPB, 0.5-1μg/ml (datasheet A02912-2); 9. Primary incubation: Overnight at 4 °C (datasheet A02912-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show very low consistency (HPA tissue IHC); Isoform / epitope: 5 isoforms; N-terminal epitopes may be lost after cleavage (UniProt); Regulation: Low expression in granulocytes (UniProt). Evidence: UniProt Q9H078, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in SK-MEL-30: 54.5 nTPM, Datasheet A02912-2, PMC11641810. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CLPB/immunohistochemistry-ihc-guide.