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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, the catalog antibody’s 0.5–1 μg/mL IHC range (datasheet A02912-2), and interpretation of the expected mitochondrial location (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); mitochondrial intermembrane space (UniProt) | |
| Staining pattern | Granular cytoplasmic staining in kidney tubule cells (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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining–RNA discordance; external verification pending (HPA tissue IHC) | |
| Regulation | Widely expressed; low in granulocytes (UniProt) | |
| Isoform / epitope | 5 isoforms; processed chains start at residues 37 or 127; epitope unknown (UniProt) |
Section 1
Recommended CLPB IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A02912-2). One published CLPB IHC protocol uses citrate pH 6.0 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 for the catalog antibody (datasheet A02912-2); the published protocol used citrate pH 6.0 with a different antibody (PMC11641810).
Section 2
What Is the Expected CLPB Staining Pattern?
CLPB resides in the mitochondrial intermembrane space and has no transmembrane segment (UniProt Q9H078). In paraffin sections, expect granular cytoplasmic staining, especially in kidney tubule cells and seminal vesicle glandular cells (HPA: general granular cytoplasmic pattern; High in both cell types). HPA rates tissue IHC reliability Enhanced, while reporting very low agreement with RNA data and pending external verification (HPA: tissue reliability).
| Strong granular cytoplasmic staining in kidney tubule cells or seminal vesicle glandular cells. | This matches the reported High staining in both cell types (HPA: tissue IHC). The granules are consistent with mitochondrial localisation, although chromogenic IHC cannot resolve the intermembrane space (UniProt Q9H078; general IHC practice). |
| Predominantly nuclear, surface, or extracellular signal with little granular cytoplasmic staining. | Treat this as a localisation mismatch and investigate artefact or nonspecific binding: CLPB is assigned to the mitochondrial intermembrane space without a transmembrane segment (UniProt Q9H078). Check the negative reagent control and whether the apparent signal follows tissue edges or pigment (general IHC practice). |
| Strong staining in cells expected to be low, while adjacent expected positive cells remain weak. | Consider cross-reactivity or endogenous chromogen activity before calling this CLPB. For example, HPA reports Low staining in hippocampal glia and Medium staining in cortical neurons (HPA: tissue IHC). Cell identity and the control pattern matter more than colour alone (general IHC practice). |
| Diffuse colour across stroma, nuclei, and multiple cell types without a granular pattern. | This does not match the reported general granular cytoplasmic profile (HPA: tissue IHC). Uneven blocking, excess antibody or detection reagent, and incomplete washes can produce broad background; assess these as general IHC workflow causes, not established CLPB-specific effects (general IHC practice). |
| No specific signal in kidney tubules or seminal vesicle glands. | These are reported High staining sites, so a blank result warrants a technical check (HPA: tissue IHC). Confirm that tissue morphology is intact, a run control developed, and primary antibody and detection reagents were applied correctly (general IHC practice). A blank slide alone does not establish absent CLPB. |
💡Expected CLPB appearanceCall a positive result when kidney tubule or seminal vesicle glandular cells show High, granular cytoplasmic colour (HPA: tissue IHC); isolated nuclear or extracellular colour is a localisation mismatch (UniProt Q9H078; general IHC practice).
| Compartment and image resolution | CLPB is in the mitochondrial intermembrane space (UniProt Q9H078). HPA describes granular cytoplasm in tissue IHC; a chromogenic section supports that pattern but cannot identify the mitochondrial subcompartment by itself (HPA: tissue IHC; general IHC practice). |
| Antibody evidence | HPA039005 and HPA039006 are IHC Enhanced; HPA064571 is IHC Approved (HPA: antibody validation). Enhanced tissue reliability has an explicit caveat: antibody staining and RNA expression show very low consistency, and external verification is pending (HPA: tissue reliability). |
| Sequence forms and epitope | UniProt lists 5 isoforms and chains beginning at residues 37 and 127 (UniProt Q9H078). Without the antibody epitope, coverage of each isoform or processed form cannot be inferred; compare the catalog antibody's stated immunogen or epitope before interpreting a missing signal. |
| IF/ICC evidence | HPA supplies no main ICC-IF location or cell line images, and lists no ICC status for these three antibodies (HPA: subcellular record; antibody validation). The tissue IHC pattern therefore does not establish an IF staining pattern or an IF protocol. |
| Retrieval and fixation evidence | No CLPB-specific retrieval condition or fixation sensitivity is supplied (UniProt Q9H078; HPA: tissue IHC). Record the retrieval conditions used and evaluate them with run controls as general IHC practice; do not attribute a change in signal to a known CLPB fixation effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| Granular signal is weak in an expected positive tissue. | The assay may have low sensitivity, or the sampled cells may differ from the reported positive population (HPA: High in kidney tubules and seminal vesicle glands). | Verify the cell type and a successful run control, then review the catalog antibody's IHC-P instructions for retrieval and dilution; make no CLPB-specific retrieval assumption (general IHC practice). |
| Colour appears mainly in nuclei or at tissue edges. | That distribution conflicts with mitochondrial localisation and the reported granular cytoplasmic tissue pattern (UniProt Q9H078; HPA: tissue IHC). | Compare matched negative reagent controls and inspect section edges and pigment; repeat staining if the distribution tracks an artefact (general IHC practice). |
| All tissue compartments show similar diffuse colour. | Broad background may reflect nonspecific binding, inadequate blocking, or incomplete washing (general IHC practice). | Review blocking, reagent concentration, wash steps, and the negative reagent control; score only cell-associated granular cytoplasm against that background (general IHC practice; HPA: tissue IHC pattern). |
| Signal persists when primary antibody is omitted. | Detection chemistry or endogenous tissue activity can create colour independently of the primary antibody (general IHC practice). | Check the primary-omission control and the detection system's endogenous-activity controls before assigning that colour to CLPB (general IHC practice). |
| A low-staining cell population appears stronger than the expected positive population. | Cross-reactivity, background, or cell misidentification is possible; HPA reports Low hippocampal glia and High kidney tubule cells (HPA: tissue IHC). | Recheck cell identity and localisation, compare controls, and seek agreement with another IHC-validated antibody where available (HPA: antibody validation; general IHC practice). |
| An IF result disagrees with the paraffin-section IHC pattern. | HPA provides no ICC-IF image-based location or ICC validation status for the listed antibodies (HPA: subcellular record; antibody validation). | Interpret the IF experiment on its own controls and documented antibody validation; do not use the IHC result as an IF protocol or subcellular confirmation (general IF 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 and expect strong staining in tubular cells (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 with less staining on the kidney slide as a background reference rather than confirmed CLPB-negative cells (HPA: no negative tissue rows).
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 slide and a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody). Use CLPB-knockout material as a biological negative where available; for kidney sections developed with biotin-based DAB detection, control endogenous peroxidase and biotin background (selected-SKU caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU tissue-IHC caption does not state its fixative (selected-SKU caption: fixative not stated). The reported paraffin-section staining used heat retrieval in EDTA at pH 8.0, so retrieval conditions should be checked when setting up kidney IHC (selected-SKU caption: heat-mediated EDTA retrieval). Whether frozen sections or IF would be easier is unestablished from the supplied evidence (HPA subcellular: no ICC-IF image cell lines); kidney staining with biotin-based DAB detection warrants background checks for endogenous biotin and peroxidase (selected-SKU caption: SABC and DAB; 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
Use the paraffin-section CLPB image as a starting point, then assess staining against mitochondrial localisation and cell-specific controls.
How should I retrieve CLPB in paraffin sections when staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin-section CLPB IHC (datasheet A02912-2). The selected image used this retrieval before incubation with 1 μg/ml catalog antibody overnight at 4°C (caption A02912-2). If staining is weak, compare retrieval times on adjacent sections while keeping antibody concentration and detection conditions constant (standard IHC practice). Include a known positive tissue, such as kidney tubules, to distinguish a retrieval problem from low local expression (HPA: High in kidney tubules). Examine whether stronger retrieval improves granular cytoplasmic signal without increasing diffuse background or tissue damage (HPA: granular cytoplasmic profile; standard IHC practice).
Can I infer the best fixative for CLPB from the paraffin IHC image?
No target-specific fixation sensitivity is established here: the selected paraffin-section image does not state its fixative (caption A02912-2). Record the fixative and fixation duration for each specimen, because those conditions affect morphology and antigen accessibility in routine IHC (standard IHC practice). When optimizing a new specimen set, compare matched sections processed with documented fixation conditions and the same pH 8.0 EDTA retrieval (datasheet A02912-2; standard IHC practice). Judge each condition using preserved tissue architecture, granular cytoplasmic staining, and a positive control processed alongside the samples (HPA: granular cytoplasmic profile; standard IHC practice). Do not assign a CLPB-specific fixation effect without a controlled comparison.
What CLPB staining pattern is plausible in chromogenic tissue IHC?
Expect predominantly granular cytoplasmic staining, consistent with CLPB localisation to the mitochondrial intermembrane space (HPA: granular cytoplasmic profile; UniProt Q9H078 localisation). CLPB has no annotated transmembrane segment, so a crisp plasma-membrane outline would require separate validation (UniProt Q9H078 topology). In kidney, inspect tubular cells as a reference population with reported High staining; breast glandular cells have reported Medium staining (HPA tissue IHC). Because chromogenic granules alone cannot establish the mitochondrial compartment, compare their distribution with a mitochondrial marker on a matched section when localisation matters (standard IHC practice). Score cell types separately so abundant unstained neighbours do not obscure a focal signal (standard IHC practice).
Could CLPB processing or isoforms explain different IHC staining patterns?
CLPB has 5 annotated isoforms and chains spanning residues 37–707 and 127–707 (UniProt Q9H078 isoforms and processing). The supplied caption does not identify the catalog antibody’s epitope, so these annotations cannot establish which forms it detects (caption A02912-2). Ask for an epitope map before interpreting loss of staining as loss of all CLPB forms, and compare matched sections with an independently characterized antibody if that distinction is essential (standard IHC practice). UniProt lists 0 glycosylation sites and an example acetylated residue at 589; neither annotation demonstrates an effect on this antibody’s staining (UniProt Q9H078 modifications). Keep processing and epitope uncertainty in the report when results differ between specimens (standard IHC practice).
How would I check a CLPB IHC pattern using multiplex IF?
Treat IF as a separate assay: the supplied catalog image documents paraffin-section chromogenic IHC, with no IF/ICC result (caption A02912-2). Pair CLPB with a marker identifying the cell population under study, such as kidney tubular cells, and a mitochondrial marker if compartment assignment is the question (HPA: High in kidney tubules; UniProt Q9H078 localisation). Choose spectrally separated fluorophores and favor a far-red channel when tissue autofluorescence interferes with shorter wavelengths (standard IF practice). Because CLPB resides in the mitochondrial intermembrane space, optimize permeabilisation for antibody access across mitochondrial membranes while monitoring loss of organelle structure (UniProt Q9H078 localisation; standard IF practice). Use single-stain and secondary-only controls to evaluate bleed-through and background (standard IF practice).
How can I separate true CLPB staining from chromogenic background?
The selected image used a biotinylated secondary antibody, streptavidin–biotin detection, and DAB, so assess background under that detection scheme (caption A02912-2). Include a peroxidase block before DAB development and a no-primary control to reveal endogenous enzyme activity or detection-reagent staining (standard IHC practice). Check whether endogenous biotin contributes signal when using a biotin-based detection system, particularly if staining persists without primary antibody (standard IHC practice). The caption used 10% goat serum for blocking and 1 μg/ml primary antibody; retain documented conditions while investigating background (caption A02912-2). Accept CLPB staining only when cellular granules exceed the control signal and tissue edges or damaged areas do not dominate (HPA: granular cytoplasmic profile; standard IHC practice).
How should I quantify CLPB staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population before scoring, because HPA reports High staining in kidney tubules and Medium staining in breast glandular cells (HPA tissue IHC). For chromogenic IHC, record the percentage of cells at each intensity and calculate an H-score from 0–300 using intensity weights 0–3 (standard IHC practice). If the question concerns abundance of positive cells, report positive-cell density per mm² of viable tissue as well (standard IHC practice). Normalize comparisons to the same cell type and viable tissue area, using consistent section thickness, imaging, and threshold settings (standard IHC practice). Keep stromal, necrotic, and target-cell measurements separate so composition changes do not masquerade as altered CLPB expression (standard IHC practice).
Which findings would make a CLPB-positive IHC result questionable?
A plausible result shows granular cytoplasmic staining in relevant cells, consistent with the HPA tissue profile and CLPB’s mitochondrial intermembrane-space localisation (HPA: granular cytoplasmic profile; UniProt Q9H078 localisation). Strong nuclear or plasma-membrane-only staining warrants scrutiny because it conflicts with that expected compartment (UniProt Q9H078 localisation and topology). Compare the claimed positive cells with a documented reference population, such as kidney tubules, while allowing for tissue-dependent expression (HPA: High in kidney tubules; UniProt Q9H078 tissue specificity). Edge-restricted staining, necrotic areas, or colour appearing in a no-primary control may indicate processing or endogenous-enzyme artefact (standard IHC practice). HPA reports very low agreement between antibody staining and RNA data despite external support, so avoid treating staining alone as definitive expression evidence (HPA reliability description).
Boster reagents
Best CLPB / Mitochondrial disaggregase IHC Antibodies
Anti-CLPB A02912-2 has IHC data from human paraffin sections of mammary and rectal cancer (catalog IHC captions) and IF data from A549 cells (catalog IF caption).
Real IHC data
Anti-CLPB Antibody ®
Cat # A02912-2
A02912-2 will render with an IHC figure from a human mammary cancer paraffin section (card image alt); another catalog caption documents IHC in a human rectal cancer paraffin section (catalog IHC caption). The same SKU lists IHC, ICC and IF applications (catalog applications) and shows IF staining in A549 cells (catalog IF caption).
Which to pick: Choose A02912-2 for tissue IHC in paraffin sections: its own figure caption reports EDTA retrieval at pH 8.0 and a 1 μg/ml primary antibody concentration in human mammary cancer tissue (A02912-2 IHC caption); the fixative is unreported (A02912-2 IHC caption). For IF/ICC, A02912-2 lists a 4 μg/ml IF concentration and shows staining in A549 cells (catalog dilution; catalog IF caption). For cross-species planning, A02912-2 lists Human, Monkey, Mouse and Rat reactivity (catalog reactivity), while its supplied IHC captions show human sections only (catalog IHC captions); clonality is unreported (catalog clone field).
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); mitochondrial intermembrane space (UniProt). 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 conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining–RNA discordance; external verification pending (HPA tissue IHC); Isoform / epitope: 5 isoforms; processed chains start at residues 37 or 127 ; epitope unknown (UniProt); Regulation: Widely expressed; low 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.