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PCCB / Propionyl-CoA carboxylase beta chain, mitochondrial · IHC design guide
Design Immunohistochemistry for PCCB
Plan PCCB chromogenic IHC on paraffin sections using the granular cytoplasmic pattern and high hepatocyte staining as references (HPA tissue IHC). Start the catalog antibody at 2–5 μg/ml (datasheet A04326-1), then assess staining against tissue controls.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PCCB 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 observed; matrix expected (HPA tissue IHC; UniProt) | |
| Staining pattern | Granular hepatocyte cytoplasm; variable across tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04326-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Bone marrow+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 | Liver peroxidase may increase chromogenic background (HPA tissue IHC; standard IHC practice) | |
| Regulation | Tissue-enhanced RNA in liver (HPA RNA specificity) | |
| Isoform / epitope | 2 isoforms; mature chain starts at residue 29; epitope map unknown (UniProt) |
Section 1
Recommended PCCB IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet A04326-1); the published osteosarcoma protocol below uses sodium citrate retrieval (PMC9751795).
| Sample | Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A04326-1) |
| Fixation | Image fixative and duration unreported (datasheet A04326-1); 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 A04326-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04326-1) |
| Primary antibody | Rabbit anti-PCCB, 2-5 μg/ml (datasheet A04326-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04326-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04326-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PCCB-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression at variable levels in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A04326-1). Sodium citrate at 95°C is the published alternative (PMC9751795).
Section 2
What Is the Expected PCCB Staining Pattern?
PCCB is a mitochondrial matrix protein with no transmembrane segment (UniProt P05166: subcellular location and topology). In paraffin IHC, expect granular cytoplasmic staining rather than a resolved view of the matrix (HPA: tissue IHC profile; general IHC interpretation). Hepatocytes and several glandular cell populations stain strongly, while expression varies across tissues (HPA: High in hepatocytes and listed glandular cells; HPA: Enhanced reliability, based on antibody staining and RNA consistency).
| Granular cytoplasmic stain in hepatocytes or listed glandular cells. | This matches the observed PCCB tissue profile (HPA: High in liver hepatocytes and adrenal, appendix, colon, duodenum, gallbladder, and parathyroid glandular cells). Granules are compatible with mitochondrial localisation, although chromogenic IHC alone does not resolve the matrix (UniProt P05166: mitochondrion matrix; general IHC interpretation). |
| Predominantly nuclear, surface-membrane, or extracellular stain. | These compartments conflict with the expected mitochondrial matrix location and granular cytoplasmic IHC profile (UniProt P05166: subcellular location and topology; HPA: tissue IHC profile). Treat an isolated compartment shift as suspect; review morphology and detection controls before assigning it to PCCB (general IHC practice). |
| Strong stain in a cell population reported as not detected. | For example, strong hematopoietic-cell staining in bone marrow differs from the HPA observation (HPA: Not detected in bone marrow hematopoietic cells). Cross-reactivity or endogenous detection activity is possible (general IHC practice). Check a no-primary control and compare the same cell population with a positive tissue; HPA's result does not prove biological absence. |
| Broad, diffuse colour obscures cell boundaries and granules. | A diffuse field cannot be scored confidently against the reported granular cytoplasmic profile (HPA: tissue IHC profile). Background may arise from detection reagents or insufficient washing (general IHC practice). Compare a no-primary control and examine whether the colour tracks tissue structures rather than the expected cell populations. |
| No convincing stain in a known-positive tissue. | Absent hepatocyte staining conflicts with the HPA High observation for liver (HPA: High in hepatocytes). First check whether the section and chromogenic detection worked, then review the antibody's IHC-P conditions (general IHC practice). A failed run cannot establish that PCCB is absent from the specimen. |
💡Expected PCCB appearanceCall a result positive when hepatocytes or listed glandular cells show convincing granular cytoplasmic signal, often strong in HPA High populations; predominantly nuclear, membranous, or uniform background colour is suspect (HPA: tissue IHC profile and High populations; UniProt P05166: mitochondrion matrix; general IHC interpretation).
| Location and topology | PCCB resides in the mitochondrial matrix and has no transmembrane segment (UniProt P05166: subcellular location and topology). Interpret chromogenic granules within cytoplasm as compatible with that location; do not infer matrix resolution from an IHC section (HPA: granular cytoplasmic profile; general IHC interpretation). |
| Tissue and cell selection | Liver hepatocytes provide a documented High-staining comparator; bone marrow hematopoietic cells are reported Not detected (HPA: liver and bone marrow tissue IHC). Use the named cell populations when comparing slides, because HPA describes variable staining across most tissues (HPA: tissue IHC profile). |
| Antibody evidence | Two listed rabbit polyclonal antibodies, HPA036939 and HPA036940, have Enhanced IHC validation (HPA: antibody validation). Enhanced supports confidence in the reported tissue pattern; it does not validate a different antibody, staining run, or specimen automatically (HPA: validation description; general IHC interpretation). |
| Processing and isoforms | UniProt lists a 29–539 mature chain and two isoforms (UniProt P05166: processing and isoforms). The supplied evidence does not map the catalog antibody's epitope or show isoform-specific tissue staining; interpret a discrepant pattern against tissue controls rather than assigning it to processing or one isoform. |
| IF/ICC Q&A: what localisation is supported? | Mitochondrial matrix localisation is annotated for PCCB (UniProt P05166: subcellular location). HPA supplies no ICC-IF images or main ICC-IF location, so its tissue IHC pattern cannot serve as direct evidence of an IF image pattern (HPA: subcellular ICC-IF record). |
| Situation | Likely cause | Next action |
|---|---|---|
| Liver control has no hepatocyte signal. | This disagrees with a documented High population (HPA: liver hepatocytes); the staining run may have failed (general IHC practice). | Check section integrity and chromogenic detection with run controls, then review the IHC-validated antibody's supplied IHC-P conditions before interpreting study tissues (general IHC practice). |
| Expected cells stain weakly, but controls develop. | Intensity varies across tissues (HPA: variable tissue IHC profile); a weak field may also be difficult to separate from background (general IHC interpretation). | Compare the named cell population with a documented High control, such as hepatocytes, on the same run; score only signal distinguishable from the no-primary control (HPA: High in hepatocytes; general IHC practice). |
| Diffuse colour appears across positive and negative regions. | Non-specific background or endogenous detection activity can obscure the reported granular pattern (general IHC practice; HPA: granular cytoplasmic tissue profile). | Inspect a no-primary control, review blocking and washing, and assess whether discrete cytoplasmic granules remain above background (general IHC practice). |
| Staining is mainly nuclear or along cell borders. | That distribution disagrees with mitochondrial matrix localisation and the tissue IHC profile (UniProt P05166: subcellular location; HPA: granular cytoplasmic profile). | Check compartment calls against the counterstain and cell morphology; repeat with appropriate run controls before scoring it as PCCB (general IHC practice). |
| A reported negative cell population stains strongly. | The result conflicts with the specified HPA observation; cross-reactivity or detection background is possible (HPA: Not detected populations; general IHC practice). | Confirm the exact cell type, compare a no-primary control, and check a documented High population in the same run before treating the signal as specific (HPA: High and Not detected populations; general IHC practice). |
| Two antibodies give conflicting cell patterns. | Both listed antibodies have Enhanced IHC status, but validation does not establish that every run will agree (HPA: HPA036939 and HPA036940 validation; general IHC interpretation). | Compare matched positive and negative cell populations, morphology, and run controls; report the discrepancy rather than combining discordant signals into one PCCB score (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PCCB staining
A04326-1 · IHCIHC analysis of PCCB using anti-PCCB antibody (A04326-1). PCCB was detected in a paraffin-embedded section of human colon adenocarcinoma tis
A04326-1 · IHCIHC analysis of PCCB using anti-PCCB antibody (A04326-1). PCCB was detected in a paraffin-embedded section of human liver cancer tissue. Hea
A04326-1 · IFIF analysis of PCCB using anti-PCCB antibody (A04326-1). PCCB was detected in a paraffin-embedded section of human liver cancer tissue. Heat
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000114054-PCCB/tissue/cerebral+cortex#imid_8890020
HPA · Salivary glandHuman Protein Atlas IHC, Salivary gland. Source: https://www.proteinatlas.org/ENSG00000114054-PCCB/tissue/salivary+gland#imid_8889984
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000114054-PCCB/tissue/colon#imid_8889954
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000114054-PCCB/tissue/liver#imid_8889947
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PCCB IHC & IF
🧪Run colon first: its glandular cells should stain for PCCB (HPA: High in colon glandular cells). Use oral mucosa squamous epithelial cells as the negative tissue (HPA: Not detected); on the colon slide, score adjacent non-glandular cells as an internal negative only where they lack specific staining.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PCCB; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a host- and clonality-matched rabbit IgG isotype control; and a biological negative such as PCCB knockout material or peptide blocking if a suitable immunogen is available (selected-SKU tissue-IHC caption: rabbit primary antibody). For chromogenic colon sections, block endogenous peroxidase and inspect glandular luminal material for nonspecific DAB signal (selected-SKU tissue-IHC caption: HRP/DAB detection in colon adenocarcinoma).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04326-1 paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption: fixative unreported). That caption reports heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required for PCCB staining (selected-SKU tissue-IHC caption: heat-mediated EDTA retrieval). No supplied evidence establishes that frozen sections or IF are easier; if using IF, assess signal against a mitochondrial-pattern expectation and tissue autofluorescence (UniProt P05166: mitochondrial matrix; standard IF practice).
HPA tissue IHC evidence for PCCB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PCCB IHC Tips
Troubleshoot chromogenic PCCB staining in paraffin sections using the catalog antibody’s tissue example, expected mitochondrial localisation, and cell-level controls.
What retrieval should I start with when PCCB staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A04326-1). The catalog antibody’s paraffin-section example used this retrieval before 10% goat-serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A04326-1). If staining remains weak, compare retrieval duration on adjacent sections while keeping antibody exposure and DAB development constant; include a no-primary control to check whether stronger treatment raises background (standard IHC practice). Judge improvement by granular cytoplasmic staining in an expected positive cell population, such as colon glandular cells, rather than by overall section darkness (HPA tissue IHC: colon glandular cells High; granular cytoplasmic profile).
Can I infer an optimal fixative for PCCB from the paraffin-section example?
No target-specific fixation sensitivity is established here: the catalog antibody’s paraffin-section caption does not state a fixative (datasheet A04326-1). Record the fixative and fixation duration for each specimen, then compare similarly processed sections when investigating weak or uneven PCCB staining (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the primary exposure at 2 μg/ml overnight at 4°C during that comparison, because those conditions are documented for the catalog example (datasheet A04326-1). Assess preserved morphology alongside granular cytoplasmic signal in expected cells; tissue staining patterns alone cannot establish a PCCB-specific fixation effect (HPA tissue IHC: granular cytoplasmic profile; standard IHC practice).
How should I assess PCCB staining that appears nuclear or along cell borders?
Expect a granular cytoplasmic pattern in chromogenic sections, consistent with PCCB’s mitochondrial-matrix localisation (HPA tissue IHC: granular cytoplasmic profile; UniProt P05166: mitochondrion matrix). PCCB has no annotated transmembrane segment, so a clean cell-border outline or predominantly nuclear deposit needs scrutiny as a staining pattern (UniProt P05166: topology; standard IHC practice). Inspect the same section for comparable deposits in cells and areas expected to stain weakly, and review a no-primary control before scoring the signal (HPA tissue IHC: variable levels; standard IHC practice). Colon glandular cells provide a documented high-staining reference, but their intensity should be judged within the same staining run (HPA tissue IHC: colon glandular cells High; standard IHC practice).
Could epitope placement or processing explain inconsistent PCCB immunostaining?
PCCB has 2 annotated isoforms, while the supplied antibody caption does not map its epitope or establish isoform recognition (UniProt P05166: isoforms; datasheet A04326-1). The annotated mature chain spans residues 29–539, and modified residues include sites at 71, 99, 248, and 474 (UniProt P05166: processing and modified residues). These annotations identify questions for an epitope review; they do not show that processing or modification changes staining with this antibody (UniProt P05166: processing and modified residues; datasheet A04326-1). When sections disagree, obtain the antibody’s immunogen or epitope information and compare matched processing conditions before assigning an isoform-specific explanation (standard IHC practice).
How can I use IF to investigate an ambiguous PCCB IHC pattern?
Use IF as a separate check of the chromogenic pattern, because the supplied catalog example documents paraffin-section IHC and the HPA payload lists no PCCB ICC/IF images (datasheet A04326-1; HPA subcellular). Multiplex PCCB with a validated marker for the cell population being assessed, such as a glandular-cell marker when investigating colon epithelium, and check whether the granular signal occurs in the expected cells (HPA tissue IHC: colon glandular cells High; standard IF practice). Choose fluorophores after checking the specimen’s autofluorescence in each channel (standard IF practice). For a mitochondrial-matrix epitope, optimise permeabilisation so antibody can reach inside mitochondria, and verify the resulting pattern with single-label and no-primary controls (UniProt P05166: mitochondrion matrix; standard IF practice).
What should I check when DAB covers the section or obscures PCCB granules?
Compare a no-primary section with the stained section to identify deposits arising from the detection system or tissue rather than primary-antibody binding (standard IHC practice). In a peroxidase/DAB workflow, include an appropriate endogenous-peroxidase block and control DAB development time; these are general chromogenic safeguards, not PCCB-specific validation (standard IHC practice). The catalog example used 10% goat serum, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary for 30 minutes at 37°C (datasheet A04326-1). If background persists, inspect whether edge staining or diffuse deposits appear in cells expected to be negative before increasing the apparent PCCB score (HPA tissue IHC: negative cell populations; standard IHC practice).
How should I quantify variable PCCB staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population before scoring, because PCCB staining varies across tissues and cell types (HPA tissue IHC: variable granular cytoplasmic expression). For a defined population, record the percentage of positive cells and an intensity-weighted H-score; if counting positive cells per mm², specify the tissue area and exclusion rules (standard IHC quantification practice). Normalise counts to evaluable tissue area, and compare H-scores only among sections processed and imaged under matched conditions (standard IHC quantification practice). Score granular cytoplasmic signal separately from diffuse background, using a documented high-staining population such as liver hepatocytes as a run reference where available (HPA tissue IHC: hepatocytes High; granular cytoplasmic profile).
When does a PCCB-positive area warrant further validation?
A convincing chromogenic result places granular signal in cytoplasm and in the expected cell population, consistent with mitochondrial-matrix PCCB and the reported tissue profile (UniProt P05166: mitochondrion matrix; HPA tissue IHC: granular cytoplasmic profile). Compare a suspected positive area with documented high-staining cells, such as colon glandular cells, and documented undetected cells, such as oral-mucosa squamous epithelium, without treating either reference as an absolute control for every specimen (HPA tissue IHC: colon High; oral mucosa Not detected; standard IHC practice). Exclude section-edge deposits and necrotic areas from scoring (standard IHC practice). Investigate nuclear-only, membrane-outline, or no-primary DAB signal as possible artefact, including endogenous-peroxidase activity, before attributing it to PCCB (UniProt P05166: mitochondrion matrix and topology; standard IHC practice).
Boster reagents
Best PCCB / Propionyl-CoA carboxylase beta chain, mitochondrial IHC Antibodies
A04326-1 has IHC images from paraffin-embedded human colon adenocarcinoma and liver cancer sections and an IF image from paraffin-embedded human liver cancer tissue (catalog image captions).
Real IHC data
Anti-PCCB Antibody ®
Cat # A04326-1
A04326-1 has documented IHC staining in paraffin-embedded human colon adenocarcinoma and liver cancer sections (catalog IHC captions). The same SKU has documented IF staining in a paraffin-embedded human liver cancer section and stated human, mouse, and rat reactivity (catalog IF caption; catalog reactivity).
Which to pick: For tissue IHC, choose A04326-1: its own captions document paraffin-section staining with EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (catalog IHC captions). For IF, the same SKU has a paraffin-section image at 5 μg/ml, while ICC validation and clonality are unreported (catalog IF caption; catalog applications and product details). For cross-species planning, it lists human, mouse, and rat reactivity, but the supplied IHC and IF tissue images show human samples only (catalog reactivity; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P05166 (PCCB_HUMAN, Propionyl-CoA carboxylase beta chain, mitochondrial).
- Human Protein Atlas. PCCB tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PCCB subcellular location (ICC-IF): Highest expression in HEL: 169.8 nTPM.
- Human Protein Atlas. PCCB antibody validation summary (2 antibodies).
- A mitochondria-related signature for predicting immune microenvironment and therapeutic response in osteosarcoma. Frontiers in oncology 2022 — PMC9751795.
- PubMed PMID:8188292 — UniProt-cited evidence.
- PubMed PMID:8225321 — UniProt-cited evidence.
- PubMed PMID:9683601 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PCCB (Propionyl-CoA carboxylase beta chain, mitochondrial). Expected localisation Granular cytoplasm observed; matrix expected (HPA tissue IHC; UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04326-1). Validated IHC antibody: A04326-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Bone marrow. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A04326-1); 2. Fixation: Image fixative and duration unreported (datasheet A04326-1); 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 A04326-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04326-1); 8. Primary antibody: Rabbit anti-PCCB, 2-5 μg/ml (datasheet A04326-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04326-1). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Liver peroxidase may increase chromogenic background (HPA tissue IHC; standard IHC practice); Isoform / epitope: 2 isoforms; mature chain starts at residue 29 ; epitope map unknown (UniProt); Regulation: Tissue-enhanced RNA in liver (HPA RNA specificity). Evidence: UniProt P05166, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in HEL: 169.8 nTPM, Datasheet A04326-1, PMC9751795. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PCCB/immunohistochemistry-ihc-guide.