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PLAT / Tissue-type plasminogen activator · IHC design guide
Design Immunohistochemistry for PLAT
Plan PLAT IHC on paraffin sections using placenta or testis as positive controls, where staining is high (HPA tissue IHC). Use the catalog antibody at 0.5–1 μg/ml (datasheet A02965-1) and score cellular and plasma staining separately (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PLAT 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 staining in a few tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in a few tissues; distinct plasma signal (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A02965-1) | |
| Positive control | Placenta+4 more · see all | |
| Negative control | Adrenal gland+4 more · see all |
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 | Secreted PLAT can appear in plasma away from producing cells (HPA tissue IHC) | |
| Regulation | No specific staining inducer is reported (UniProt) | |
| Isoform / epitope | 4 isoforms; cleavage into A/B chains makes epitope location relevant (UniProt) |
Section 1
Recommended PLAT IHC & IF Protocols
The catalog antibody protocol uses citrate pH 6 heat retrieval (datasheet: A02965-1). One published PLAT IHC protocol specifies a tissue sample and secondary detection (PMC9366818).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet A02965-1) |
| Fixation | Image fixative and duration unreported (datasheet A02965-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A02965-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02965-1) |
| Primary antibody | Rabbit anti-PLAT, 0.5-1μg/ml (datasheet A02965-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02965-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02965-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PLAT-positive staining in syncytiotrophoblasts - cell body of placenta (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a few tissues. Distinct positivity in plasma. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6 heat retrieval (datasheet: A02965-1); the published protocol does not state a retrieval method (PMC9366818).
Section 2
What Is the Expected PLAT Staining Pattern?
PLAT is secreted into extracellular space and has no transmembrane segment (UniProt P00750). In paraffin tissue sections, expect selected cell-body staining alongside distinct plasma positivity: HPA reports high staining in placental syncytiotrophoblasts and testicular spermatogonia, with medium staining in several other cell types (HPA tissue IHC). HPA rates its tissue IHC evidence Supported and cautions that a secreted protein’s RNA and protein locations may differ (HPA tissue IHC).
| Strong cell-body staining in placental syncytiotrophoblasts or testicular spermatogonia, with distinct plasma positivity. | This matches the reported high-staining cells and plasma pattern (HPA tissue IHC). Score the named cells separately from plasma: extracellular PLAT can be present away from its site of synthesis (UniProt P00750; HPA tissue IHC). |
| Strong nuclear-only staining, or a sharp membrane rim presented as the principal PLAT pattern. | Treat this as unexpected and investigate an artifact before scoring it as PLAT: UniProt places PLAT in extracellular space and lists no transmembrane segment (UniProt P00750). HPA’s tissue description instead reports limited cytoplasmic staining and distinct plasma positivity (HPA tissue IHC). |
| Prominent staining in cells reported as undetected, such as adrenal glandular cells or colon endothelial cells. | Possible cross-reactivity or endogenous chromogen-producing activity warrants investigation; the named cells are reported as Not detected (HPA tissue IHC). Compare a no-primary control and the same run’s positive tissue before assigning specificity (standard IHC practice). |
| A uniform haze covers cells, stroma and empty areas, obscuring the reported cell pattern. | Diffuse background cannot establish PLAT localisation. HPA describes distinct plasma positivity, which should be judged separately from a uniform slide-wide deposit (HPA tissue IHC). Check no-primary background and detection chemistry, then reassess identifiable cells (standard IHC practice). |
| No signal in placental syncytiotrophoblasts or testicular spermatogonia on the run’s positive section. | A negative study section is inconclusive until the positive section works: both cell populations have high reported staining (HPA tissue IHC). Review the antibody’s IHC validation and the run’s retrieval, dilution and detection conditions (HPA antibody validation; standard IHC practice). |
💡Expected PLAT appearanceCall a positive result when identifiable syncytiotrophoblast or spermatogonia cell bodies show high staining, with distinct plasma positivity where present; isolated strong nuclear staining or uniform slide-wide color is suspect (HPA tissue IHC; UniProt P00750; standard IHC practice).
| Secretion and compartment | PLAT is secreted and lacks a transmembrane segment (UniProt P00750). HPA reports cytoplasmic staining in a few tissues and distinct plasma positivity; do not require every positive deposit to remain inside its producing cell (HPA tissue IHC). |
| Antibody validation | HPA rates tissue IHC reliability Supported and lists two antibodies with Supported IHC status, HPA003412 and CAB009335 (HPA tissue IHC; HPA antibodies). That status supports comparison with the reported pattern but does not establish that every stained cell is specific. |
| Processing and antibody epitope | PLAT has a cleaved signal sequence, propeptides, and mature chain A and chain B regions (UniProt P00750). If an antibody’s epitope is known, check its location before interpreting absent staining; no epitope or resulting IHC sensitivity is supplied here. |
| IF/ICC: should actin-filament signal be expected? | HPA reports an approved actin-filament location in ICC-IF images from A-431, U-251MG and U2OS (HPA subcellular). This is a separate assay observation; it does not replace the secreted location or establish an actin-filament pattern in paraffin tissue IHC (UniProt P00750; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive control has no cell-body signal. | The expected high-staining cells may have been missed, or the IHC run may have failed (HPA tissue IHC; standard IHC practice). | Confirm the section contains syncytiotrophoblasts or spermatogonia, then inspect the antibody’s validated IHC conditions and detection controls (HPA tissue IHC; standard IHC practice). |
| The whole section is brown, including expected negative areas. | Nonspecific background or endogenous detection activity may obscure the reported limited cytoplasmic and distinct plasma pattern (HPA tissue IHC; standard IHC practice). | Run a no-primary control; review blocking, washes and chromogen development before changing the PLAT interpretation (standard IHC practice). |
| Strong staining appears in adrenal glandular cells or colon endothelial cells. | These cells are reported as Not detected, so cross-reactivity or endogenous activity is possible (HPA tissue IHC; standard IHC practice). | Compare with a no-primary control and a known-positive section in the same run; report the unexpected cell type separately (HPA tissue IHC; standard IHC practice). |
| Plasma stains but expected positive cell bodies do not. | PLAT secretion permits extracellular signal apart from cellular signal, while HPA reports both plasma positivity and high staining in selected cells (UniProt P00750; HPA tissue IHC). | Score plasma and cells separately; verify tissue identity and the IHC run before calling the cell result negative (HPA tissue IHC; standard IHC practice). |
| Only strong nuclear or continuous membrane-rim staining is seen. | That distribution conflicts with PLAT’s secreted, non-transmembrane annotation and HPA’s limited cytoplasmic tissue pattern (UniProt P00750; HPA tissue IHC). | Check the no-primary control and compare with the named positive cells; withhold a PLAT-specific call until the unexpected pattern is resolved (standard IHC practice; HPA tissue IHC). |
| A negative call depends on one antigen-retrieval setting. | The supplied UniProt and HPA records give no PLAT-specific retrieval condition or fixation-sensitivity finding. | Check the catalog antibody’s IHC-P instructions and confirm that a reported high-staining cell population works in the same run before interpreting the study section (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing PLAT staining
A02965-1 · IHCIHC analysis of PLAT using anti-PLAT antibody (A02965-1). PLAT was detected in paraffin-embedded section of human placenta tissue. Heat medi
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000104368-PLAT/tissue/cerebral+cortex#imid_1406252
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000104368-PLAT/tissue/duodenum#imid_1406169
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000104368-PLAT/tissue/colon#imid_1406170
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000104368-PLAT/tissue/liver#imid_1406187
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003412; green = PLAT, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003412; green = PLAT, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PLAT IHC & IF
🧪Run placenta first: syncytiotrophoblast cell bodies show High PLAT staining (HPA: High in placenta syncytiotrophoblasts). Use adrenal gland glandular cells as a negative comparison (HPA: Not detected in adrenal gland glandular cells); on the placenta slide, use unstained adjacent cells as an internal background reference without assuming they are PLAT-negative.
Positive control tissue: Placenta (Syncytiotrophoblasts - cell body, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PLAT in A-431, U-251MG, U2OS, with annotated localisation: Actin filaments (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit isotype controls (selected-SKU caption: rabbit primary antibody; standard IHC practice), plus PLAT-knockout material as a biological specificity control (standard IHC practice). Check endogenous peroxidase and biotin background in placenta before interpreting DAB signal (selected-SKU caption: biotin-based DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption). That caption uses citrate retrieval at pH 6 for 20 minutes, but does not establish that retrieval is required (selected-SKU caption). No supplied comparison establishes whether frozen sections or IF are easier; secreted PLAT may produce extracellular staining that needs careful interpretation, and placental endogenous biotin or peroxidase can complicate chromogenic scoring (UniProt P00750: secreted, extracellular space; selected-SKU caption: biotin-based DAB detection; standard IHC practice).
HPA tissue IHC evidence for PLAT
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Secreted protein, tissue location of RNA and protein is expected to differ. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Placenta | Syncytiotrophoblasts - cell body | High | Protein (IHC) | HPA → |
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PLAT IHC Tips
Troubleshoot PLAT staining in paraffin sections by checking retrieval, tissue controls and compartment specific signal before comparing intensity across samples.
What retrieval conditions should I start with if PLAT staining is weak?
Start with heat mediated retrieval in citrate buffer at pH 6 (datasheet A02965-1). The catalog antibody stained a paraffin embedded human placenta section after 20 minutes of this retrieval, followed by incubation at 1 μg/ml overnight at 4°C (datasheet A02965-1). If staining remains weak, check that the section stayed covered with buffer during heating and compare retrieval times on adjacent sections while holding detection conditions constant (standard IHC practice). Include placenta as a positive control and inspect syncytiotrophoblast cell bodies, where high PLAT staining has been reported (HPA: High in placenta syncytiotrophoblasts).
Could fixation explain weak or uneven PLAT staining?
The catalog image shows staining in a paraffin embedded placenta section, but its caption does not state the fixative (datasheet A02965-1). PLAT specific sensitivity to any fixative or fixation duration is therefore unknown from the supplied evidence (datasheet A02965-1: fixative not stated). Record each sample’s fixative and fixation time, then compare adjacent sections processed with the same retrieval and detection conditions (standard IHC practice). If signal differs across batches, check tissue preservation and use a shared positive control before attributing the difference to PLAT expression (standard IHC practice).
Should I expect PLAT staining inside cells or in extracellular material?
Assess both cellular and extracellular staining: PLAT is secreted into the extracellular space and has no transmembrane segment (UniProt P00750: subcellular location and topology). Cytoplasmic staining in a few tissues and distinct plasma positivity are reported, so an extracellular signal alone does not identify the producing cell (HPA: tissue IHC profile and reliability description). In placenta, compare staining with syncytiotrophoblast cell bodies, which show high positivity in the tissue profile (HPA: High in placenta syncytiotrophoblasts). Document cellular and extracellular compartments separately, using section morphology and a negative control to judge whether diffuse chromogen reflects specific deposition or background (standard IHC practice).
Can this stain distinguish PLAT isoforms or processed chains?
Do not assign a stained structure to one PLAT isoform without antibody epitope and isoform coverage data; the record lists 4 isoforms (UniProt P00750: isoforms). PLAT also undergoes signal peptide and propeptide removal, and its mature protein contains disulfide linked A and B chains (UniProt P00750: processing and subunit). Its fibronectin type I, EGF like, kringle and peptidase domains offer distinct potential epitope regions, while glycosylation is recorded at 4 sites (UniProt P00750: domains and glycosylation). Treat the chromogenic result as antibody reactivity in the section until the reagent’s epitope and cross isoform recognition are established (standard IHC interpretation).
How should I investigate PLAT localisation by multiplex IF?
On the separate IF/ICC workflow, pair PLAT with an independently validated marker for the expected cell type, such as syncytiotrophoblasts when studying placenta (HPA: High in placenta syncytiotrophoblasts; standard IF practice). Choose spectrally separated fluorophores and check an unstained section for tissue autofluorescence, favoring a longer wavelength channel when background is high (standard IF practice). PLAT is secreted and lacks a transmembrane segment, but the catalog antibody’s epitope location is unspecified (UniProt P00750: location and topology; datasheet A02965-1: epitope not stated). Test permeabilised and nonpermeabilised preparations to distinguish accessible extracellular staining from intracellular signal, and use single label controls to check channel bleed through (standard IF practice).
How can I reduce diffuse or vessel associated chromogenic background?
First compare stained sections with a no primary control and inspect the distribution of chromogen at the same exposure and microscope settings (standard IHC practice). PLAT is secreted, binds fibrin, and can occur in plasma, so vessel or extracellular staining needs interpretation rather than automatic rejection (UniProt P00750: location, fibrin binding and tissue specificity; HPA: plasma positivity). The catalog placenta example used 10% goat serum blocking, 1 μg/ml primary antibody and biotin based DAB detection (datasheet A02965-1). If background persists, check endogenous peroxidase and biotin contributions, then adjust blocking, washing or antibody concentration while retaining the positive control (standard IHC practice).
How should I score PLAT staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring because PLAT is secreted and HPA reports both cytoplasmic staining and plasma positivity (UniProt P00750: location; HPA: tissue IHC profile). For cell associated signal, report the percentage of positive cells and an intensity based H-score of 0–300, with scoring thresholds fixed before comparing groups (standard IHC practice). For extracellular signal, quantify positive area or optical density within a defined tissue area, normalised to area in mm² (standard image analysis practice). Keep retrieval, chromogen development, imaging and region selection consistent, and analyse placenta control sections alongside each staining batch (datasheet A02965-1: placenta staining; standard IHC practice).
Which staining patterns support a true PLAT positive result?
A credible result follows interpretable tissue structures and is reproducible in a positive control, such as placenta syncytiotrophoblast cell bodies (HPA: High in placenta syncytiotrophoblasts; standard IHC practice). Extracellular or plasma associated signal can be plausible because PLAT is secreted and binds fibrin, but it cannot by itself establish which cell produced the protein (UniProt P00750: location and fibrin binding; HPA: plasma positivity). Review unexpected nuclear staining, strong section edges and necrotic areas against the no primary control before calling them positive (standard IHC practice). Also check whether residual endogenous peroxidase or biotin could explain DAB signal in the catalog antibody’s biotin based detection workflow (datasheet A02965-1; standard IHC practice).
Boster reagents
Best PLAT / Tissue-type plasminogen activator IHC Antibodies
The catalog includes human-, mouse-, and rat-reactive anti-PLAT antibodies listed for IHC (catalog: reactivity and applications). A human placenta paraffin-section image documents IHC staining with A02965-1 (A02965-1 image caption).
Real IHC data
Anti-TPA Tissue Plasminogen Activator/PLAT Antibody ®
Cat # A02965-1
A02965-1 has an IHC image from a human placenta paraffin section and is listed for IHC, IHC-F, and ICC in human, mouse, and rat (A02965-1 image caption; catalog: applications and reactivity). M02965 is listed for IHC in human, mouse, and rat, with no IHC image supplied (catalog: M02965 applications, reactivity, and image alts).
Which to pick: For paraffin-section IHC, choose A02965-1 when an illustrated starting condition is useful: its caption reports citrate retrieval at pH 6 for 20 minutes and 1 μg/ml primary antibody; the fixative is unreported (A02965-1 image caption). For ICC, A02965-1 is the listed choice; no IF image is supplied (catalog: applications and IF image alts). For cross-species IHC, both SKUs list human, mouse, and rat reactivity; M02965 is the rabbit monoclonal option, while A02965-1 has the human placenta paraffin-section image (catalog: reactivity and clone; A02965-1 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P00750 (TPA_HUMAN, Tissue-type plasminogen activator).
- Human Protein Atlas. PLAT tissue IHC expression (reliability: Supported).
- Human Protein Atlas. PLAT subcellular location (ICC-IF): Localized to the actin filaments..
- Human Protein Atlas. PLAT antibody validation summary (2 antibodies).
- Mesenteric Lymphatic-Perilymphatic Adipose Crosstalk: Role in Alcohol-Induced Perilymphatic Adipose Tissue Inflammation. Alcoholism, clinical and experimental research 2015 — PMC5102000.
- Red blood cell distribution width is associated with increased interactions of blood cells with vascular wall. Scientific reports 2022 — PMC9366818.
- Extracellular matrix-associated gene expression in adult sensory neuron populations cultured on a laminin substrate. BMC neuroscience 2013 — PMC3610289.
- PubMed PMID:6337343 — UniProt-cited evidence.
- PubMed PMID:6089198 — UniProt-cited evidence.
- PubMed PMID:3009482 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PLAT (Tissue-type plasminogen activator). Expected localisation Cytoplasmic staining in a few tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A02965-1). Validated IHC antibody: A02965-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Placenta (Syncytiotrophoblasts - cell body); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human placenta tissue; fixative not specified (datasheet A02965-1); 2. Fixation: Image fixative and duration unreported (datasheet A02965-1); 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: Citrate pH 6 , 20 min (datasheet A02965-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02965-1); 8. Primary antibody: Rabbit anti-PLAT, 0.5-1μg/ml (datasheet A02965-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02965-1). Watch for: Fixation: Keep fixation conditions consistent across sections (standard IHC practice; not target-specific); Caveat: Secreted PLAT can appear in plasma away from producing cells (HPA tissue IHC); Isoform / epitope: 4 isoforms; cleavage into A/B chains makes epitope location relevant (UniProt); Regulation: No specific staining inducer is reported (UniProt). Evidence: UniProt P00750, HPA tissue IHC · Supported, HPA ICC-IF · actin filaments., Datasheet A02965-1, PMC9366818. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PLAT/immunohistochemistry-ihc-guide.