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PLP2 / Proteolipid protein 2 · IHC design guide
Design Immunohistochemistry for PLP2
Plan PLP2 paraffin-section IHC using the reported cytoplasmic tissue pattern (HPA tissue IHC). The catalog antibody has an IHC dilution range of 2–5 μg/ml (datasheet A06255-1); interpret staining in light of the low consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PLP2 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 | General cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in glandular cells and hepatocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06255-1) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Ovary |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06255-1) | |
| Caveat | Antibody staining and RNA show low consistency (HPA tissue IHC) | |
| Regulation | A4 mRNA peaks in lower colonic crypts (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage and orientation are unspecified (UniProt) |
Section 1
Recommended PLP2 IHC & IF Protocols
The catalog antibody protocol uses heat-mediated EDTA retrieval (datasheet A06255-1). The published PLP2 IHC protocols below provide tissue staining conditions (PMC7077595; PMC10642424; PMC6274732; PMC8113544).
| Sample | Paraffin-embedded human adenocarcinoma of the right colon tissue; fixative not specified (datasheet A06255-1) |
| Fixation | Image fixative and duration unreported (datasheet A06255-1); 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 A06255-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06255-1) |
| Primary antibody | Rabbit anti-PLP2, 2-5 μg/ml (datasheet A06255-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06255-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06255-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PLP2-positive staining in purkinje cells of cerebellum (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A06255-1). When reproducing a published protocol, use its stated retrieval conditions (PMC7077595; PMC6274732; PMC8113544).
Section 2
What Is the Expected PLP2 Staining Pattern?
PLP2 is a four-pass membrane protein (UniProt Q04941 topology). In paraffin-section IHC, expect staining in cell populations reported by HPA, including duodenal glandular cells and kidney tubule cells, both scored High (HPA tissue IHC). HPA describes the tissue pattern as generally cytoplasmic and rates the IHC antibody Approved, while noting low agreement with RNA data and pending external verification (HPA tissue IHC).
| Distinct staining in duodenal glandular cells or kidney tubule cells, with a cytoplasmic or membrane-associated appearance. | This fits HPA High staining in those cells and its general cytoplasmic IHC profile (HPA tissue IHC). Membrane association is consistent with PLP2's four transmembrane segments (UniProt Q04941 topology). Assess signal in the named cells rather than treating every cell in a positive tissue as an expected positive. |
| Predominantly nuclear staining, with little staining in the expected cells. | A predominantly nuclear pattern conflicts with the membrane annotation and reported cytoplasmic tissue pattern (UniProt Q04941 subcellular location; HPA tissue IHC). Treat it as a possible artefact and review detection and controls (general IHC practice); morphology alone cannot identify its cause. |
| Strong staining outside the reported positive cell population, such as in ovarian stroma cells. | HPA reports ovarian stroma cells as Not detected (HPA tissue IHC). Unexpected signal raises possible cross-reactivity or endogenous detection activity (general IHC practice). Check cell identity and appropriate controls before assigning either cause; HPA's low antibody–RNA agreement limits certainty (HPA tissue IHC). |
| A uniform haze or stain across cells and surrounding section, obscuring cell boundaries. | This is difficult to score as cell-specific PLP2 staining (general IHC practice). Review background in a no-primary control, blocking, washing and detection conditions (general IHC practice). HPA's general cytoplasmic profile does not make uniform slide-wide colour a convincing positive pattern (HPA tissue IHC). |
| No visible signal in the named positive cells of an otherwise interpretable section. | Absence in HPA High cells, such as duodenal glandular cells, warrants a run-control check (HPA tissue IHC; general IHC practice). Confirm that the cells are present, then review antibody, retrieval and detection steps (general IHC practice). A failed run does not establish that the specimen lacks PLP2. |
💡Expected PLP2 appearanceCall a result positive when the named HPA High cells show clear cytoplasmic or membrane-associated staining; widespread nuclear colour, uniform haze or staining confined to unexpected cells is suspect (HPA tissue IHC; UniProt Q04941 topology; general IHC practice).
| Membrane topology and IHC appearance | PLP2 has four transmembrane segments and a MARVEL domain spanning residues 19–137 (UniProt Q04941). HPA nevertheless summarizes tissue IHC as generally cytoplasmic (HPA tissue IHC). Interpret apparent cytoplasmic colour alongside the expected cell population; the supplied sources do not specify a finer organelle pattern. |
| Choice of comparison cells | HPA scores Purkinje cells, duodenal and gallbladder glandular cells, kidney tubule cells, hepatocytes, small-intestinal glandular cells and Leydig cells High (HPA tissue IHC). Ovarian stroma cells are Not detected (HPA tissue IHC). These are cell-specific comparisons, not whole-organ pass or fail labels. |
| Intestinal distribution | UniProt reports enrichment in colonic mucosa and the strongest A4 message in the lower half of the crypt along the crypt–villus axis (UniProt Q04941 tissue specificity). This describes message distribution; it does not establish an IHC intensity gradient. HPA separately reports High glandular-cell staining in duodenum and small intestine (HPA tissue IHC). |
| Antibody evidence | The listed rabbit polyclonal antibody, HPA042415, is Approved for IHC, with no ICC status listed (HPA antibodies). HPA describes low consistency between antibody staining and RNA expression and says external verification is pending (HPA tissue IHC). Use its staining profile as a reference that needs controls, not as definitive specificity proof. |
| IF/ICC Q: What localisation should be expected? | A: HPA's subcellular summary says Membrane, but it lists no main location or cell-line ICC-IF images (HPA subcellular). The listed antibody has no ICC status (HPA antibodies). The supplied evidence therefore supports a broad membrane expectation, without an image-backed cell-line pattern or an IF/ICC protocol recommendation. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known HPA High cells appear unstained. | The run, tissue sampling or identification of the expected cells may need checking (general IHC practice). | Locate the named cells and compare a known-positive section processed in the same run; review the antibody, retrieval and detection steps (HPA tissue IHC; general IHC practice). Do not infer a PLP2-specific retrieval requirement from these sources. |
| Colour fills nuclei more strongly than cytoplasm. | The pattern conflicts with HPA's general cytoplasmic IHC description and UniProt's membrane annotation (HPA tissue IHC; UniProt Q04941). | Check the no-primary control and inspect morphology and counterstain before scoring the result (general IHC practice). If nuclear colour persists, record it as discordant rather than calling it expected PLP2 localisation. |
| Diffuse colour obscures expected cells. | Nonspecific background or detection-related colour may be contributing (general IHC practice). | Compare a no-primary control, then review blocking, washing and detection conditions (general IHC practice). Score PLP2 only where cell-specific staining can be separated from background; HPA's cytoplasmic profile alone cannot resolve an obscured slide (HPA tissue IHC). |
| Ovarian stroma cells stain strongly. | That cell population is Not detected in HPA tissue IHC; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Verify the cell population and compare no-primary and detection controls (general IHC practice). Treat the discrepancy as a specificity question, especially given HPA's reported low antibody–RNA agreement (HPA tissue IHC). |
| The chosen comparison tissue gives only weak staining. | The comparison cells may belong to an HPA Low group, such as parathyroid glandular or bronchial respiratory epithelial cells (HPA tissue IHC). | Compare the same run with named HPA High cells before judging sensitivity (HPA tissue IHC; general IHC practice). Record the cell type and its expected HPA level, since a weak Low-category comparator cannot serve the same purpose as a High-category one. |
| Intestinal staining does not follow a clear crypt gradient. | UniProt's lower-crypt gradient concerns A4 message, while HPA reports glandular-cell protein staining (UniProt Q04941 tissue specificity; HPA tissue IHC). | Score the observed glandular-cell staining and document its position without requiring an RNA-like intensity gradient (HPA tissue IHC; general IHC practice). Check controls if the named HPA High cells lack signal altogether. |
Experiment evidence
IHC & IF images showing PLP2 staining
A06255-1 · IHCIHC analysis of PLP2 using anti-PLP2 antibody (A06255-1). PLP2 was detected in a paraffin-embedded section of human adenocarcinoma of the ri
A06255-1 · IHCIHC analysis of PLP2 using anti-PLP2 antibody (A06255-1). PLP2 was detected in a paraffin-embedded section of human cervical cancer tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000102007-PLP2/tissue/cerebral+cortex#imid_11189069
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000102007-PLP2/tissue/colon#imid_11187501
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000102007-PLP2/tissue/liver#imid_11187494
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000102007-PLP2/tissue/kidney#imid_11187503
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PLP2 IHC & IF
🧪Run duodenum first and expect staining in glandular cells (HPA: High in duodenal glandular cells). Use ovary as the negative tissue, focusing on ovarian stroma cells (HPA: Not detected in ovarian stroma cells); on the duodenal slide, assess adjacent nonglandular areas as background comparators rather than assuming they lack PLP2.
Positive control tissue: Cerebellum (Purkinje cells, HPA High)
Negative control tissue: Ovary (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PLP2; derive a cell-line control from the positive tissue's cell type (Purkinje cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody). Use PLP2 knockout tissue or peptide competition, where available, as a biological specificity control; block endogenous peroxidase before HRP–DAB detection and inspect the duodenal lumen for nonspecific precipitate (selected-SKU caption: HRP–DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU tissue-IHC caption does not state the fixative (selected-SKU caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used for a paraffin-embedded colon adenocarcinoma section, but retrieval dependence in duodenum is unreported (selected-SKU caption). There is no supplied frozen-section or IF/ICC result establishing an easier route (HPA subcellular: no ICC-IF image cell lines).
HPA tissue IHC evidence for PLP2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PLP2 IHC Tips
Use the paraffin-section example as a starting point for PLP2 IHC, then verify staining against tissue architecture and appropriate controls.
Which antigen retrieval conditions should I try first for PLP2 paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A06255-1). The selected PLP2 tissue image used that condition, followed by overnight primary-antibody incubation at 4°C (caption A06255-1). Keep heating and cooling conditions consistent across comparison sections, and include a section processed without primary antibody to assess detection background (standard IHC practice). If staining is weak, vary retrieval duration on adjacent sections before testing another buffer, since excessive retrieval can damage tissue morphology and make membrane-associated staining difficult to judge (standard IHC practice; UniProt Q04941 topology).
How should I assess whether fixation is affecting PLP2 staining?
The selected paraffin-section caption does not state a fixative, so PLP2-specific sensitivity to fixation is unknown (caption A06255-1). Record the fixative and fixation duration for each specimen, then compare sections processed with the same retrieval, antibody concentration, and detection conditions (standard IHC practice). Check whether weak signal accompanies poor preservation of tissue architecture or varies with fixation history; neither observation alone establishes a PLP2-specific fixation effect (standard IHC practice). Use a consistently processed positive tissue section in each run, and interpret differences cautiously because fixation can alter antigen accessibility in paraffin IHC (standard IHC practice).
Should PLP2 appear at the membrane or throughout the cytoplasm?
PLP2 is annotated as a membrane protein with 4 transmembrane segments, spanning residues 25–45, 48–68, 85–105, and 112–132 (UniProt Q04941 topology). HPA describes its tissue IHC profile as general cytoplasmic expression, creating a localisation discrepancy that deserves validation (HPA tissue IHC). Assess whether DAB signal follows plausible cell boundaries or intracellular membrane-rich regions at high magnification, while comparing adjacent negative-control sections (UniProt Q04941 topology; standard IHC practice). Diffuse staining across unrelated compartments should prompt a review of retrieval, antibody concentration, and background before it is assigned to PLP2 (standard IHC practice).
Can an unknown antibody epitope limit interpretation of PLP2 IHC?
PLP2 has 2 annotated isoforms, but the supplied product caption does not identify the antibody epitope or establish isoform recognition (UniProt Q04941 isoforms; caption A06255-1). Its 4 membrane-spanning segments and glycosylation sites at residues 18 and 108 make epitope accessibility a relevant question, without predicting this antibody’s behaviour (UniProt Q04941 topology and glycosylation). Check the antibody’s documented immunogen and isoform coverage before assigning differences in staining to splice variants (standard IHC practice). If those details remain unavailable, report the result as antibody-detected PLP2 staining and validate any isoform-specific interpretation independently (standard IHC practice).
How can I extend the PLP2 IHC findings to multiplex immunofluorescence?
For a separate IF experiment, pair PLP2 with a validated marker of the expected glandular cell population; HPA reports high PLP2 staining in duodenal glandular cells (HPA tissue IHC). Choose fluorophores in channels with low measured tissue autofluorescence, and inspect single-stain controls before interpreting overlap (standard IF practice). PLP2 crosses the membrane 4 times, but the supplied evidence does not place this antibody’s epitope on a particular side of the membrane (UniProt Q04941 topology; caption A06255-1). Establish epitope orientation before selecting permeabilisation conditions, then compare permeabilised and nonpermeabilised controls without treating the paraffin IHC result as IF validation (standard IF practice).
What should I change if PLP2 DAB staining is widespread?
The selected paraffin-section example used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and HRP with DAB detection (caption A06255-1). Begin by checking a no-primary control and applying a peroxidase block to assess endogenous enzyme signal; these are general chromogenic IHC controls (standard IHC practice). If background persists, titrate the primary antibody around the documented concentration and verify that washes remove unbound reagent without lifting tissue (caption A06255-1; standard IHC practice). Judge any improvement against preserved cell detail, since PLP2’s membrane annotation alone cannot establish that diffuse DAB staining is specific (UniProt Q04941 subcellular annotation; standard IHC practice).
How should I quantify PLP2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then record the percentage of positive cells and intensity on a 0–3 scale (standard IHC practice). An H-score sums each intensity category multiplied by its percentage, yielding 0–300; use the same threshold and imaging conditions across sections (standard IHC practice). Normalise positive-cell counts to the number of evaluable cells, or report positive-cell density per mm² of viable tissue when cell counting is impractical (standard IHC practice). Keep crypt position in the region annotation because PLP2 message was reported as most abundant in the lower half of colonic crypts (UniProt Q04941 tissue specificity).
What findings would support a specific PLP2 IHC signal?
A credible result combines reproducible cellular staining with low signal in a no-primary control and a plausible membrane-associated pattern (UniProt Q04941 subcellular annotation; standard IHC practice). Examine the stained cell type carefully: HPA reports high staining in several populations, including duodenal glandular cells, but flags low consistency between antibody staining and RNA expression (HPA tissue IHC). Exclude section-edge accentuation, necrotic areas, and endogenous peroxidase signal before calling DAB-positive cells true positives (standard IHC practice). Treat disagreement between cytoplasmic IHC staining and PLP2’s membrane annotation as a reason for additional validation, since neither observation alone establishes specificity (HPA tissue IHC; UniProt Q04941 subcellular annotation).
Boster reagents
Best PLP2 / Proteolipid protein 2 IHC Antibodies
A06255-1 has real IHC data from human paraffin sections (A06255-1 image captions). The catalog provides no IF/ICC figure or nonhuman reactivity (A06255-1 catalog).
Real IHC data
Anti-PLP2 Antibody ®
Cat # A06255-1
A06255-1 is listed for human IHC and shows staining in paraffin sections of right colon adenocarcinoma and cervical cancer (A06255-1 catalog and image captions). Its other IHC captions show human gastric carcinoma and liver cancer; no IF image is supplied (A06255-1 image captions and catalog).
Which to pick: Choose A06255-1 for human paraffin-section IHC: its own captions report EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml (A06255-1 image captions); the fixative is unreported (A06255-1 image captions). No listed SKU is validated for IF/ICC or nonhuman samples, so this catalog provides no supported pick for either (A06255-1 applications and reactivity). Clonality is unspecified (A06255-1 catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q04941 (PLP2_HUMAN, Proteolipid protein 2).
- Human Protein Atlas. PLP2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PLP2 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. PLP2 antibody validation summary (1 antibodies).
- Reduced expression of proteolipid protein 2 increases ER stress-induced apoptosis and autophagy in glioblastoma. Journal of cellular and molecular medicine 2020 — PMC7077595.
- PLP2 Could Be a Prognostic Biomarker and Potential Treatment Target in Glioblastoma Multiforme. Pharmacogenomics and personalized medicine 2023 — PMC10642424.
- Proteolipid Protein 2 Overexpression Indicates Aggressive Tumor Behavior and Adverse Prognosis in Human Gliomas. International journal of molecular sciences 2018 — PMC6274732.
- CircRNA_2646 functions as a ceRNA to promote progression of esophageal squamous cell carcinoma via inhibiting miR-124/PLP2 signaling pathway. Cell death discovery 2021 — PMC8113544.
- PubMed PMID:8470895 — UniProt-cited evidence.
- PubMed PMID:9344658 — UniProt-cited evidence.
- PubMed PMID:16344560 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PLP2 (Proteolipid protein 2). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06255-1). Validated IHC antibody: A06255-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Ovary. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human adenocarcinoma of the right colon tissue; fixative not specified (datasheet A06255-1); 2. Fixation: Image fixative and duration unreported (datasheet A06255-1); 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 A06255-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06255-1); 8. Primary antibody: Rabbit anti-PLP2, 2-5 μg/ml (datasheet A06255-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06255-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06255-1); Caveat: Antibody staining and RNA show low consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage and orientation are unspecified (UniProt); Regulation: A4 mRNA peaks in lower colonic crypts (UniProt). Evidence: UniProt Q04941, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A06255-1, PMC7077595, PMC10642424. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PLP2/immunohistochemistry-ihc-guide.