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PLS1 / Plastin-1 · IHC design guide
Design Immunohistochemistry for PLS1
Plan chromogenic PLS1 IHC on paraffin sections using colon glandular cells as a high-staining reference (HPA tissue IHC). This guide covers fixation, retrieval, a 2–5 μg/ml antibody range (datasheet A07675-1), and scoring of observed membranous and cytoplasmic staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PLS1 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 | Observed membranous and cytoplasmic staining, mainly in GI tract (HPA tissue IHC) | |
| Staining pattern | GI glandular cells show membranous and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07675-1) | |
| 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 fixation consistent across paraffin sections and controls. (selected-SKU IHC image A07675-1) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Intestine-enriched expression (HPA tissue IHC) | |
| Isoform / epitope | No isoforms; one cytoplasmic 1–629 chain, with no extracellular region (UniProt) |
Section 1
Recommended PLS1 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet: A07675-1); the published colorectal cancer tissue-core protocol below provides an additional example (PMC7419044).
| Sample | Paraffin-embedded human colon tissue; fixative not specified (datasheet A07675-1) |
| Fixation | Image fixative and duration unreported (datasheet A07675-1); 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 A07675-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07675-1) |
| Primary antibody | Rabbit anti-PLS1, 2-5 μg/ml (datasheet A07675-1) |
| Primary incubation | Overnight at 4 °C (datasheet A07675-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07675-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PLS1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Membranous and cytoplasmic expression was mainly observed in gastrointestinal tract. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A07675-1); the published protocol reports antigen retrieval without its conditions (PMC7419044 methods).
Section 2
What Is the Expected PLS1 Staining Pattern?
PLS1 is cytoplasmic and associates with cell projections; it has no transmembrane segment (UniProt Q14651). On paraffin sections, expect mainly membranous and cytoplasmic staining in gastrointestinal glandular cells (HPA tissue IHC). HPA rates the tissue pattern Enhanced, while noting only medium consistency between antibody staining and RNA expression (HPA tissue IHC). Interpret an apical rim as a staining pattern, without treating it as proof that PLS1 spans the membrane (UniProt Q14651 topology).
| Strong glandular-cell staining in small intestine, duodenum, colon, rectum, or appendix, with cytoplasmic and membranous emphasis. | This fits the reported gastrointestinal pattern and High glandular-cell staining in each listed tissue (HPA tissue IHC). Assess signal against adjacent unstained structures on the same slide; uniform chromogen over every structure is less convincing as a cell-specific result (general IHC practice). |
| Predominantly nuclear staining, with little cytoplasmic or membranous signal. | This conflicts with the reported tissue pattern (HPA tissue IHC) and cytoplasmic, cell-projection localization (UniProt Q14651). Review the no-primary control, counterstain, and detection background before assigning nuclear localization; a nuclear-only result is a warning sign, not evidence of a new compartment (general IHC practice). |
| Strong signal in a cell population reported as undetected, such as adipocytes in adipose tissue. | HPA reports PLS1 as Not detected in those adipocytes (HPA tissue IHC). Consider nonspecific antibody binding or endogenous detection activity; compare a no-primary control and an independently processed known-positive section before interpreting the unexpected cells (general IHC practice). |
| Diffuse chromogen across glandular cells, stroma, and empty spaces, without clear cellular boundaries. | That distribution does not resemble HPA's mainly gastrointestinal glandular-cell pattern (HPA tissue IHC). Broad background can reflect nonspecific detection, insufficient blocking, or overly concentrated antibody; check controls and optimize the general IHC workflow before scoring PLS1 (general IHC practice). |
| No convincing signal in small-intestinal or colonic glandular cells. | Both are High in HPA tissue IHC, so a blank section calls for a technical check before a biological negative call (HPA tissue IHC). Confirm tissue identity and section integrity, then review the antibody dilution, retrieval conditions, and detection controls (general IHC practice). |
💡Expected PLS1 appearanceCall a section positive when glandular cells show clear, often High cytoplasmic and membranous staining in the listed gastrointestinal tissues (HPA tissue IHC); isolated nuclear signal or uniform tissue-wide chromogen is suspect (UniProt Q14651 localization; general IHC practice).
| Cell and tissue distribution | HPA reports High glandular-cell staining in appendix, colon, duodenum, gallbladder, rectum, and small intestine; Sertoli cells in testis are also High, while bronchial respiratory epithelium is Medium (HPA tissue IHC). Choose controls by the stated cell type, since a whole tissue contains multiple populations (general IHC practice). |
| Compartment and topology | UniProt places PLS1 in cytoplasm and stereocilia and reports no transmembrane segment (UniProt Q14651). HPA describes mainly membranous and cytoplasmic gastrointestinal staining (HPA tissue IHC). A membrane-adjacent pattern is compatible with these reports, but chromogenic IHC alone cannot establish membrane insertion (general IHC practice). |
| Strength of the tissue evidence | The HPA tissue profile is Enhanced, and two listed antibodies have Enhanced IHC status: HPA055744 and HPA061936 (HPA tissue IHC; HPA antibodies). HPA also reports medium consistency between antibody staining and RNA expression, so an unexpected result deserves control-based review (HPA tissue IHC). |
| IF/ICC: what localization should be expected? | HPA reports mainly plasma-membrane localization in ICC-IF, but cautions that this assessment uses antibodies targeting proteins from multiple genes (HPA subcellular). Compare that observation with UniProt's cytoplasmic and stereocilium annotations (UniProt Q14651); the ICC-IF entry is a localization clue, not an IHC protocol (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive intestinal glandular cells are blank. | A failed staining or detection step is possible when a High HPA cell population lacks signal (HPA tissue IHC; general IHC practice). | Verify tissue identity, inspect the positive-control section, and check primary-antibody dilution, retrieval, and detection reagents against the selected IHC method (general IHC practice). No PLS1-specific retrieval condition is supplied here. |
| Only nuclei stain. | The compartment differs from HPA's membranous/cytoplasmic tissue profile and UniProt's cytoplasmic/cell-projection assignment (HPA tissue IHC; UniProt Q14651). | Compare the no-primary control and counterstain; repeat with controlled detection conditions before calling the nuclear signal PLS1 (general IHC practice). |
| Adipocytes in adipose tissue stain strongly. | HPA lists these cells as Not detected, so nonspecific binding or endogenous detection activity should be considered (HPA tissue IHC; general IHC practice). | Inspect a no-primary control and the chosen detection-system control, then compare with a glandular-cell positive control on the same run (general IHC practice; HPA tissue IHC). |
| Chromogen spreads across multiple unrelated tissue compartments. | Diffuse background obscures the mainly glandular-cell pattern reported by HPA (HPA tissue IHC); excess primary antibody or incomplete background suppression are general possibilities (general IHC practice). | Check the no-primary control, optimize blocking and primary-antibody dilution, and score only signal that resolves to identifiable cells (general IHC practice). |
| A low-signal tissue looks negative. | Cholangiocytes in liver and urothelial cells in urinary bladder are reported at Low levels, so a faint or absent result there is less decisive than in a High control (HPA tissue IHC). | Run a listed High glandular-cell control and compare the specified cell populations; avoid declaring assay failure from the low-level tissue alone (HPA tissue IHC; general IHC practice). |
| An unexpected cell pattern persists after basic controls pass. | HPA's Enhanced profile still has only medium antibody-to-RNA consistency, leaving some discordance unresolved by the tissue summary alone (HPA tissue IHC). | Document the cell type and compartment, compare the pattern with HPA's tissue map, and seek independent antibody or orthogonal support before reporting a new distribution (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PLS1 staining
A07675-1 · IHCIHC analysis of PLS1 using anti-PLS1 antibody (A07675-1). PLS1 was detected in a paraffin-embedded section of human colon tissue. Heat media
A07675-1 · IHCIHC analysis of PLS1 using anti-PLS1 antibody (A07675-1). PLS1 was detected in a paraffin-embedded section of human colon cancer tissue. Hea
A07675-1 · IFIF analysis of PLS1 using anti-PLS1 antibody (A07675-1). PLS1 was detected in an immunocytochemical section of A549 cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000120756-PLS1/tissue/cerebral+cortex#imid_20902727
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000120756-PLS1/tissue/colon#imid_20902645
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000120756-PLS1/tissue/liver#imid_20902654
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000120756-PLS1/tissue/kidney#imid_20902655
HPA · ICC-IF JURKATHuman Protein Atlas immunofluorescence, JURKAT, antibody HPA000895; green = PLS1, blue = nucleus, red = microtubules.
HPA · ICC-IF JURKATHuman Protein Atlas immunofluorescence, JURKAT, antibody HPA000895; green = PLS1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PLS1 IHC & IF
🧪Run colon first and expect staining in its glandular cells (HPA: colon glandular cells High). Run adipose tissue as the negative tissue, focusing on adipocytes (HPA: adipocytes Not detected); on the colon slide, use adjacent nonglandular cells as a background reference without assuming they are PLS1-negative.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PLS1 in JURKAT, SiHa, U2OS, with annotated localisation: Plasma membrane (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit isotype controls alongside a validated PLS1 knockout biological negative (caption: rabbit primary antibody). Block endogenous peroxidase in colon sections and check the no-primary slide for DAB background (caption: colon tissue, HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU colon caption does not state a fixative (caption: A07675-1). The reported paraffin-section result used heat retrieval in EDTA at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C (caption: A07675-1); the supplied evidence does not establish whether frozen sections or IF/ICC are easier. In colon, endogenous peroxidase can contribute to chromogenic background, so assess it with the no-primary control (standard IHC practice).
HPA tissue IHC evidence for PLS1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Rectum | 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 → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PLS1 IHC Tips
Troubleshoot PLS1 staining in paraffin sections by checking retrieval, cell type, compartment and controls before comparing signal intensity (UniProt Q14651; HPA tissue IHC).
What should I change if PLS1 staining is weak after antigen retrieval?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin-section PLS1 staining (datasheet A07675-1). Compare a colon control processed alongside the test section, because the catalog antibody detected PLS1 in paraffin-embedded human colon (datasheet A07675-1). Keep section thickness, heating conditions and detection reagents consistent while assessing whether glandular-cell staining improves (standard IHC practice; HPA tissue IHC). If signal remains weak, optimize retrieval heating on matched sections and inspect morphology before changing the buffer (standard IHC practice). The caption reports retrieval conditions but does not report a fixative, so it cannot establish fixation-specific retrieval needs (datasheet A07675-1).
Can I infer how fixation affects PLS1 staining in paraffin sections?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state a fixative (datasheet A07675-1). Record the fixative and processing history for each specimen, then compare sections with a matched colon control under identical staining conditions (standard IHC practice; datasheet A07675-1). Keep heat-mediated EDTA retrieval at pH 8.0 as the starting condition for the catalog antibody (datasheet A07675-1). If signal differs across specimens, assess tissue preservation and retrieval consistency before assigning the difference to PLS1 abundance (standard IHC practice). Neither the reported gastrointestinal staining pattern nor PLS1 topology establishes a fixation effect (HPA tissue IHC; UniProt Q14651).
Where should convincing PLS1 staining appear in a colon section?
Look for staining in glandular cells of colon, where PLS1 is reported at a high level (HPA tissue IHC). Cytoplasmic and membrane-associated patterns are plausible because gastrointestinal IHC shows both, while UniProt places PLS1 in cytoplasm and cell projections (HPA tissue IHC; UniProt Q14651). PLS1 has no transmembrane segment, so a membrane-adjacent signal should be assessed as localisation rather than evidence of membrane insertion (UniProt Q14651). Check that staining follows intact cell outlines or cytoplasm across the gland, rather than pooling over folds or tissue edges (standard IHC practice). Use the colon section demonstrated for the catalog antibody as a practical comparison (datasheet A07675-1).
How should I assess an unexpected PLS1 staining pattern without a mapped epitope?
The supplied PLS1 record lists 0 isoforms and a single chain spanning residues 1–629 (UniProt Q14651). It identifies two EF-hand and four calponin-homology domains, but the selected antibody caption does not locate its epitope (UniProt Q14651; datasheet A07675-1). Therefore, do not assign a compartment-specific staining difference to epitope masking or isoform recognition without additional antibody evidence (standard IHC practice). Compare retrieval-matched sections and cell-type distribution with the documented colon glandular pattern before interpreting a new signal (datasheet A07675-1; HPA tissue IHC). A missing mapped epitope also prevents a domain-level explanation for variable staining (datasheet A07675-1; UniProt Q14651).
How can I adapt the PLS1 assessment to multiplex IF?
Treat IF as a separate validation task: the selected antibody caption documents chromogenic paraffin-section IHC, without supplying an IF protocol (datasheet A07675-1). Pair PLS1 with an independently validated glandular-cell marker when examining intestinal tissue, where glandular-cell signal is reported (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and check unstained tissue for autofluorescence before assigning a weak PLS1 channel (standard IF practice). Because PLS1 is cytoplasmic and has no transmembrane segment, compare mild permeabilisation with an unpermeabilised condition if intracellular access matters (UniProt Q14651; standard IF practice). The antibody epitope and IF fixation sensitivity are unspecified, so optimize both experimentally (datasheet A07675-1).
How do I distinguish diffuse DAB background from PLS1 signal?
First check whether brown signal resolves to glandular cells and their cytoplasmic or membrane-associated regions in gastrointestinal tissue (HPA tissue IHC). The selected colon example used 10% goat-serum blocking and DAB chromogen, providing a documented starting workflow for this antibody (datasheet A07675-1). Include a no-primary control to assess secondary-reagent and chromogen background, and use a peroxidase block when evaluating endogenous enzyme activity (standard IHC practice). Compare background across tissue edges, folds and intact glands before increasing antibody concentration or development time (standard IHC practice). If background persists, adjust blocking, washing or detection conditions one variable at a time (standard IHC practice).
How should I score PLS1 IHC across colon specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define glandular cells as the primary scoring compartment because colon glandular cells show high PLS1 staining (HPA tissue IHC). For comparable sections, record the percentage of positive glandular cells and an H-score based on their staining intensity (standard IHC practice). Normalize positive-cell counts to the total evaluable glandular-cell count, or report positive area relative to evaluable glandular area (standard IHC practice). Apply the same thresholds, retrieval and DAB development conditions to every specimen, with a shared colon control to monitor staining variation (standard IHC practice; datasheet A07675-1). Exclude folds, damaged edges and poorly preserved regions from the predefined analysis area (standard IHC practice).
When should I question an apparent PLS1-positive area?
Give greatest weight to reproducible staining in intact gastrointestinal glandular cells, where the reported PLS1 pattern is cytoplasmic and membranous (HPA tissue IHC). A signal confined to nuclei or unrelated cell populations warrants further checking against PLS1's cytoplasmic and cell-projection localisation (UniProt Q14651; HPA tissue IHC). Inspect abrupt edge staining, folds and necrotic regions before treating their DAB deposits as expression (standard IHC practice). Run a no-primary control and assess peroxidase blocking when brown deposits appear outside expected cells (standard IHC practice). Interpret weak or discordant staining cautiously because HPA reports only medium consistency between antibody staining and RNA expression (HPA tissue IHC).
Boster reagents
Best PLS1 / Plastin-1 IHC Antibodies
A07675-1 has IHC images of paraffin-embedded human colon, colon cancer and stomach cancer, plus an IF image of A549 cells (catalog image captions).
Real IHC data
Anti-PLS1 Antibody ®
Cat # A07675-1
A07675-1 is listed for IHC and IF/ICC, with images of paraffin-embedded human colon, colon cancer and stomach cancer sections (catalog applications; IHC captions). Its IF image shows A549 cells, and the catalog lists Human, Mouse and Rat reactivity (catalog IF caption; catalog reactivity).
Which to pick: For tissue IHC, choose A07675-1: its paraffin-section captions document EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (catalog IHC captions). For IF/ICC, the same SKU is listed for both applications, with an A549 IF image using 5 μg/ml primary antibody (catalog applications; catalog IF caption). For cross-species planning, A07675-1 lists Human, Mouse and Rat reactivity, but the supplied IHC images show human tissue and the IF image shows A549 cells; clonality is unreported (catalog reactivity; catalog image captions; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14651 (PLSI_HUMAN, Plastin-1).
- Human Protein Atlas. PLS1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PLS1 subcellular location (ICC-IF): Mainly localized to the plasma membrane. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. PLS1 antibody validation summary (3 antibodies).
- Plastin 1 drives metastasis of colorectal cancer through the IQGAP1/Rac1/ERK pathway. Cancer science 2020 — PMC7419044.
- PubMed PMID:8139549 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PLS1 (Plastin-1). Expected localisation Observed membranous and cytoplasmic staining, mainly in GI tract (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07675-1). Validated IHC antibody: A07675-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon tissue; fixative not specified (datasheet A07675-1); 2. Fixation: Image fixative and duration unreported (datasheet A07675-1); 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 A07675-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07675-1); 8. Primary antibody: Rabbit anti-PLS1, 2-5 μg/ml (datasheet A07675-1); 9. Primary incubation: Overnight at 4 °C (datasheet A07675-1). Watch for: Fixation: Keep fixation consistent across paraffin sections and controls. (selected-SKU IHC image A07675-1); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No isoforms; one cytoplasmic 1–629 chain, with no extracellular region (UniProt); Regulation: Intestine-enriched expression (HPA tissue IHC). Evidence: UniProt Q14651, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. Caution: Based on…, Datasheet A07675-1, PMC7419044. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PLS1/immunohistochemistry-ihc-guide.