This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
PLXDC1 / Plexin domain-containing protein 1 · IHC design guide
Design Immunohistochemistry for PLXDC1
Plan PLXDC1 chromogenic IHC on paraffin sections using its observed cytoplasmic and membranous tissue pattern (HPA tissue IHC). Start the catalog antibody at 2.5 μg/mL (datasheet), and assess glandular and endothelial staining by cell type (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PLXDC1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic with membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic, sometimes membranous, in several cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+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 | Glandular staining can complicate endothelial scoring (HPA tissue IHC) | |
| Regulation | Tumour endothelium enriched versus normal (UniProt) | |
| Isoform / epitope | 4 isoforms; verify the epitope’s isoform and membrane side (UniProt) |
Section 1
Recommended PLXDC1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published gastric cancer protocol (PMC9488617).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A08584-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-PLXDC1, 2.5 μg/mL (datasheet A08584-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PLXDC1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with a membranous pattern in several tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); the published pH 7.8 method is an alternative (PMC9488617).
Section 2
What Is the Expected PLXDC1 Staining Pattern?
PLXDC1 is a transmembrane protein with extracellular and cytoplasmic regions; UniProt places it predominantly at vascular endothelial tight junctions and, to a lesser extent, the luminal surface (UniProt Q8IUK5 topology and subcellular location). On tissue sections, HPA also reports general cytoplasmic staining and membranous staining in several tissues, including high staining in specified glandular and hematopoietic cells (HPA tissue IHC). HPA rates its tissue IHC profile Approved, with medium consistency against RNA data (HPA tissue IHC).
| Vascular cell borders stain, with weaker luminal staining. | This matches UniProt's predominant tight-junction and lesser luminal localization in vascular endothelial cells (UniProt Q8IUK5 subcellular location). Interpret borders in their tissue context: HPA also describes general cytoplasmic and some membranous staining outside vessels (HPA tissue IHC). |
| Glandular cells show cytoplasmic staining, sometimes with a membrane pattern. | That can fit the HPA profile: glandular cells are High in appendix, epididymis, gallbladder and stomach (HPA tissue IHC). Do not require every positive cell to show a crisp endothelial-junction pattern; compare its compartment with the HPA profile (HPA tissue IHC). |
| A strong, predominantly nuclear pattern appears in the cells being scored. | A nuclear pattern does not match the listed membrane, junction, cytoplasm or secreted locations (UniProt Q8IUK5 subcellular location; HPA tissue IHC). Check the no-primary control and staining distribution before calling it PLXDC1 (general IHC practice). |
| Unexpected cells stain while the expected cells in the same section do not. | Check the exact cell population against HPA, which reports both positive and undetected populations within named tissues (HPA tissue IHC). If the pattern remains discordant, investigate antibody cross-reactivity or endogenous detection activity with controls (general IHC practice). |
| A known HPA-positive cell population has no discernible signal. | High HPA staining identifies a useful comparator, such as appendix glandular cells or bone-marrow hematopoietic cells (HPA tissue IHC). Absence in one run calls for checking controls and assay performance before interpreting the specimen as negative (general IHC practice). |
💡Expected PLXDC1 appearanceCall a positive result when staining follows plausible endothelial junction or luminal, cytoplasmic, or tissue-specific membranous patterns in the relevant cells, with High HPA comparator populations visibly stained; a dominant nuclear pattern or uniform staining of unrelated cells is suspect (UniProt Q8IUK5 subcellular location; HPA tissue IHC; general IHC practice).
| Cell population and tissue | Use cell-level comparators: HPA calls appendix glandular cells and bone-marrow hematopoietic cells High, but adipocytes and cardiomyocytes Not detected (HPA tissue IHC). Whole-section positivity cannot substitute for identifying which cells stain (general IHC practice). |
| Endothelial context | UniProt reports PLXDC1 in endothelial cells from several primary cancers and not detectable in normal-tissue endothelial cells (UniProt Q8IUK5 tissue specificity). A negative normal vessel therefore cannot by itself invalidate a run; assess a suitable positive comparator (general IHC practice). |
| Topology and compartment | Residues 19–426 are extracellular, 427–447 span the membrane, and 448–500 are cytoplasmic (UniProt Q8IUK5 topology). The record gives no antibody epitope, so it cannot predict which region the catalog antibody recognizes (UniProt Q8IUK5 topology; HPA antibody record). |
| Processing and protein forms | UniProt lists a signal peptide at 1–18, a 19–500 chain, four isoforms and glycosylation sites at 33, 80 and 197 (UniProt Q8IUK5). These annotations do not establish an IHC-specific staining difference or a shedding mechanism (UniProt Q8IUK5). |
| Strength of HPA evidence | The tissue IHC profile is Approved with medium antibody–RNA consistency; the listed antibody HPA012805 is IHC Approved (HPA tissue IHC; HPA antibody record). Treat an unusual pattern as a finding to check, rather than assuming antibody-independent confirmation (HPA tissue IHC; HPA antibody record). |
| IF/ICC: is a membrane signal established? | HPA summarizes PLXDC1 as membrane-localized but provides no ICC-IF images; HPA012805 has no listed ICC status (HPA subcellular; HPA antibody record). Use that as a localization clue, not a validated IF/ICC result or protocol (HPA subcellular; HPA antibody record). |
| Situation | Likely cause | Next action |
|---|---|---|
| A positive-control section has little or no staining. | A failed staining run is possible; HPA reports High staining in appendix glandular cells and bone-marrow hematopoietic cells (HPA tissue IHC). | Verify the positive comparator's cell identity, then check retrieval, primary antibody dilution, detection reagents and counterstain using routine IHC controls (general IHC practice). No PLXDC1-specific retrieval setting is supplied (HPA tissue IHC; UniProt Q8IUK5). |
| A normal vessel is negative, but a cancer vessel stains. | That contrast is compatible with UniProt's report of detection in cancer endothelial cells and nondetection in normal-tissue endothelial cells (UniProt Q8IUK5 tissue specificity). | Score the endothelial cells in each specimen separately and check a positive tissue comparator before treating the normal-vessel result as technical failure (UniProt Q8IUK5 tissue specificity; general IHC practice). |
| Most of the section has diffuse brown background. | Nonspecific antibody binding or detection background may obscure the cell-level pattern (general IHC practice). | Inspect no-primary and detection-only controls, review blocking and washes, and compare the result with HPA's cell-specific distribution (general IHC practice; HPA tissue IHC). |
| An undetected HPA cell population appears strongly positive. | A discordant pattern may reflect cross-reactivity or endogenous detection activity; HPA lists adipocytes and cardiomyocytes as Not detected (HPA tissue IHC; general IHC practice). | Confirm the cell identity, examine no-primary and detection-only controls, and repeat with appropriate blocking or an independent antibody if available (general IHC practice). |
| Staining is mainly nuclear, with no plausible membrane or cytoplasmic signal. | The compartment conflicts with UniProt's membrane, junction and cytoplasmic locations and HPA's cytoplasmic or membranous tissue profile (UniProt Q8IUK5 subcellular location; HPA tissue IHC). | Check the counterstain and no-primary control, then reassess staining conditions and antibody specificity before scoring nuclei as positive (general IHC practice). |
| IF/ICC shows a membrane-like signal, but its specificity is uncertain. | HPA's membrane summary has no accompanying ICC-IF images, and the listed antibody has no ICC validation status (HPA subcellular; HPA antibody record). | Evaluate IF/ICC on its separate guide page with appropriate controls; do not treat IHC approval as IF/ICC validation (HPA antibody record; general IF/ICC practice). |
Experiment evidence
IHC & IF images showing PLXDC1 staining
A08584-1 · IHCImmunohistochemistry of TEM7 in mouse liver tissue with TEM7 antibody at 2.5 μg/mL.
A08584-1 · IFImmunofluorescence of TEM7 in mouse liver tissue with TEM7 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000161381-PLXDC1/tissue/cerebral+cortex#imid_3358712
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000161381-PLXDC1/tissue/colon#imid_3358638
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000161381-PLXDC1/tissue/liver#imid_3358639
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000161381-PLXDC1/tissue/gallbladder#imid_3358615
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PLXDC1 IHC & IF
🧪Run appendix first: glandular cells should stain (HPA: High in appendix glandular cells). Use heart muscle as the negative tissue, where cardiomyocytes are not detected (HPA: heart muscle cardiomyocytes Not detected); on the appendix slide, adjacent cells without specific staining should remain at background, with their status assessed empirically.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PLXDC1; 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 no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus PLXDC1-knockout tissue as a biological negative (standard IHC practice). Quench endogenous peroxidase for chromogenic detection, especially when evaluating inflammatory cells in appendix (standard IHC practice).
⚠️Feasibility: No PLXDC1-specific fixation window or antigen-retrieval effect is reported in the supplied evidence; optimize retrieval empirically for paraffin sections. The A08584-1 mouse-liver IHC caption does not report a fixative (catalog caption: fixative not stated). The payloads do not establish whether frozen sections or IF/ICC are easier; in appendix, distinguish glandular staining from peroxidase-associated background in inflammatory cells (HPA: High in appendix glandular cells; standard IHC practice).
HPA tissue IHC evidence for PLXDC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cilia axoneme) | High | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PLXDC1 IHC Tips
Use compartment-aware controls and matched section processing to troubleshoot PLXDC1 staining in chromogenic paraffin-section IHC (UniProt Q8IUK5 topology and localisation; standard IHC practice).
How should I retrieve PLXDC1 in paraffin sections when junctional staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 minutes (this page’s antigen-retrieval method). Cool sections in buffer, then compare staining with a matched section processed without heat retrieval to assess whether retrieval improves the signal (standard IHC practice). Score endothelial junctions separately from diffuse cytoplasm because PLXDC1 is reported predominantly at vascular endothelial tight junctions, with lesser luminal staining (UniProt Q8IUK5 localisation). If signal remains weak, test a different retrieval buffer or pH on adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Document any increase in tissue damage or background alongside the gain in specific staining (standard IHC practice).
Could fixation explain weak or uneven PLXDC1 staining?
PLXDC1-specific sensitivity to fixation is unknown from the supplied evidence; neither the mouse liver caption nor the tissue staining summary establishes a fixation condition (catalog antibody A08584-1 caption; HPA tissue IHC). Record the actual fixative, fixation duration, section thickness and processing history for each paraffin block before comparing staining (standard IHC practice). Compare adjacent sections with identical citrate pH 6.0 retrieval at 95–98 °C for 20 minutes and the same detection settings (this page’s antigen-retrieval method; standard IHC practice). Include a section known to stain under the same run conditions to distinguish a block-specific problem from a run-wide failure (standard IHC practice). Do not interpret differences between blocks as a PLXDC1-specific fixation effect without a controlled fixation comparison (standard IHC practice).
Which staining compartments should count as plausible PLXDC1 signal?
Inspect vascular endothelial borders for junction-associated staining and the luminal surface for weaker staining, the reported predominant and lesser locations respectively (UniProt Q8IUK5 localisation). PLXDC1 has an extracellular region at residues 19–426, a transmembrane segment at 427–447 and a cytoplasmic region at 448–500 (UniProt Q8IUK5 topology). Record membranous and cytoplasmic staining separately because tissue IHC also reports general cytoplasmic expression with a membranous pattern in several tissues (HPA tissue IHC). Do not assign every stained cell to endothelium: glandular cells in appendix and gallbladder are reported as High (HPA tissue IHC). Resolve uncertain cell identity by morphology and a serial-section cell-type marker before scoring vascular signal (standard IHC practice).
How can epitope position affect interpretation of PLXDC1 staining?
First obtain the antibody’s mapped immunogen or epitope before claiming that staining represents a particular PLXDC1 region or isoform (standard IHC practice). The supplied record lists 4 isoforms but gives no isoform-specific epitope map, so this antibody cannot distinguish them on that evidence alone (UniProt Q8IUK5 isoforms; catalog antibody A08584-1 caption). PLXDC1 contains an extracellular region at residues 19–426 and a cytoplasmic region at 448–500, separated by a transmembrane segment (UniProt Q8IUK5 topology). Glycosylation is annotated at residues 33, 80 and 197, but its effect on this antibody’s IHC signal is unestablished (UniProt Q8IUK5 glycosylation; catalog antibody A08584-1 caption). If results disagree across antibodies, compare documented epitope locations and staining compartments on adjacent sections before inferring an isoform difference (standard IHC practice).
How should I investigate PLXDC1 localisation with a separate IF experiment?
For a separate IF/ICC experiment, multiplex PLXDC1 with a validated endothelial marker to test whether junctional signal follows the expected vascular cell population (UniProt Q8IUK5 localisation; standard IF practice). Choose spectrally separated fluorophores after checking unstained tissue autofluorescence; favor a channel with low autofluorescence for the weaker target signal (standard IF practice). Determine whether the antibody binds the extracellular residues 19–426 or cytoplasmic residues 448–500 before choosing permeabilisation: cytoplasmic access generally requires it, whereas excessive permeabilisation can complicate surface-pattern interpretation (UniProt Q8IUK5 topology; standard IF practice). Titrate permeabilisation on matched samples and include single-label and secondary-only controls when assessing colocalisation (standard IF practice). The supplied HPA subcellular record lists no ICC/IF images, so it offers no direct image check for this experiment (HPA subcellular).
What should I check when PLXDC1 staining appears diffuse or widespread?
Run a no-primary control and inspect unstained areas before assigning diffuse brown signal to PLXDC1; endogenous peroxidase and nonspecific detection can produce chromogenic background (standard IHC practice). Apply a peroxidase block and protein block according to the detection workflow, and keep DAB development time matched across sections (standard IHC practice). Compare each section with a tissue or cell compartment expected to have little staining, such as adipocytes reported as Not detected in adipose tissue (HPA tissue IHC). The catalog image reports mouse liver staining at 2.5 µg/mL, but its caption does not state fixation or establish that concentration for other samples (catalog antibody A08584-1 caption). If background persists, titrate antibody concentration and review washing and tissue-edge staining before changing interpretation thresholds (standard IHC practice).
How should I quantify PLXDC1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring: record endothelial junctional or luminal signal separately from glandular and diffuse cytoplasmic signal (UniProt Q8IUK5 localisation; HPA tissue IHC). For a chosen cell population, report the percentage of positive cells and an intensity-based H-score using 0–3 intensity categories, with the same threshold across slides (standard IHC practice). For vascular patterns, an alternative is positive-vessel density per mm², normalised to the total vessels or vascular area assessed in the same region (standard IHC practice). Exclude folds, necrotic regions and section edges by prespecified rules, and sample comparable viable areas across specimens (standard IHC practice). Report region selection, denominator, scorer agreement and control performance because HPA assigns its tissue IHC profile Approved reliability with medium staining–RNA consistency (HPA tissue IHC; standard IHC practice).
How do I distinguish credible PLXDC1 staining from artefact?
Give greatest weight to reproducible vascular junctional staining, with possible lesser luminal staining, because those are reported PLXDC1 locations in endothelial cells (UniProt Q8IUK5 localisation). Evaluate other positive cell populations by identity rather than rejecting them solely for lacking a vascular pattern: HPA reports High staining in appendix glandular cells and bone-marrow hematopoietic cells (HPA tissue IHC). Question signal confined to a section edge, necrotic area or no-primary control, and inspect endogenous peroxidase before calling DAB deposition positive (standard IHC practice). Diffuse nuclear-only staining warrants scrutiny because the supplied localisation records identify membrane, junctional and cytoplasmic locations, without a nuclear assignment (UniProt Q8IUK5 localisation; HPA tissue IHC). Interpret negative endothelium cautiously across settings because tumour-associated endothelial detection and a low cerebral-cortex endothelial IHC signal come from different observations (UniProt Q8IUK5 tissue specificity; HPA tissue IHC).
Boster reagents
Best PLXDC1 / Plexin domain-containing protein 1 IHC Antibodies
A08584-1 has mouse liver IHC and IF images (catalog image captions) and lists Human, Mouse, and Rat reactivity (catalog reactivity).
Real IHC data
Anti-TEM7 PLXDC1 Antibody
Cat # A08584-1
A08584-1 is listed for IHC-P and IF (catalog applications). Its IHC and IF images show mouse liver tissue at 2.5 μg/mL and 20 μg/mL, respectively (catalog image captions).
Which to pick: Choose A08584-1 for paraffin-section IHC: IHC-P is listed, and its own mouse liver IHC image uses 2.5 μg/mL (catalog applications; A08584-1 IHC image caption). The same SKU is listed for IF and has a mouse liver IF image at 20 μg/mL; ICC is not documented (catalog applications; A08584-1 IF image caption). It lists Human, Mouse, and Rat reactivity, but the pictured IHC and IF samples are mouse liver; the IHC caption does not report a fixative (catalog reactivity; A08584-1 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IUK5 (PLDX1_HUMAN, Plexin domain-containing protein 1).
- Human Protein Atlas. PLXDC1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PLXDC1 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. PLXDC1 antibody validation summary (1 antibodies).
- PLXDC1 Can Be a Biomarker for Poor Prognosis and Immune Evasion in Gastric Cancer. Journal of inflammation research 2022 — PMC9488617.
- Selective delivery of PLXDC1 small interfering RNA to endothelial cells for anti-angiogenesis tumor therapy using CD44-targeted chitosan nanoparticles for epithelial ovarian cancer. Drug delivery 2018 — PMC6096458.
- Plexin domain-containing 1 may be a biomarker of poor prognosis in hepatocellular carcinoma patients, may mediate immune evasion. World journal of gastrointestinal oncology 2024 — PMC11099457.
- PubMed PMID:10947988 — UniProt-cited evidence.
- PubMed PMID:11559528 — UniProt-cited evidence.
- PubMed PMID:15574754 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PLXDC1 (Plexin domain-containing protein 1). Expected localisation Cytoplasmic with membranous tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A08584-1 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A08584-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-PLXDC1, 2.5 μg/mL (datasheet A08584-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Glandular staining can complicate endothelial scoring (HPA tissue IHC); Isoform / epitope: 4 isoforms; verify the epitope’s isoform and membrane side (UniProt); Regulation: Tumour endothelium enriched versus normal (UniProt). Evidence: UniProt Q8IUK5, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A08584-1, PMC9488617. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PLXDC1/immunohistochemistry-ihc-guide.