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SRPK1 / SRSF protein kinase 1 · IHC design guide
Design Immunohistochemistry for SRPK1
This guide maps expected cytoplasmic SRPK1 staining in paraffin sections (HPA tissue IHC). Use appendix glandular cells as a high-staining reference (HPA tissue IHC) and the IHC-validated antibody at 5 μg/ml (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SRPK1 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 | Predominantly cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Broad cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01937-1) | |
| Caveat | Stress or Hsp90 inhibition can shift SRPK1 into nuclei (UniProt) | |
| Regulation | Isoform 2 predominates in testis (UniProt) | |
| Isoform / epitope | 3 isoforms; verify epitope coverage across variants (UniProt) |
Section 1
Recommended SRPK1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by three published SRPK1 IHC protocols (PMC4351909; PMC3842442; PMC9609466).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A01937-1); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-SRPK1, 5 μg/ml (datasheet A01937-1) |
| 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 | SRPK1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval recommendation); assess cytoplasmic and nuclear staining (UniProt Q96SB4 localization).
Section 2
What Is the Expected SRPK1 Staining Pattern?
SRPK1 staining in paraffin sections should be widespread and predominantly cytoplasmic, including strong staining in glandular, respiratory epithelial and neuronal cells (HPA: ubiquitous cytoplasmic expression; High in the listed cells). Nuclear signal can occur because SRPK1 shuttles between cytoplasm and nucleus (UniProt Q96SB4: subcellular location). SRPK1 has no transmembrane segment (UniProt Q96SB4: topology). HPA rates the tissue IHC profile Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC reliability).
| Cytoplasmic staining is strong in colon or breast glandular cells, or bronchial respiratory epithelial cells. | This matches reported high staining in those cell types (HPA: High in colon and breast glandular cells; High in bronchial respiratory epithelial cells). Score the cells and compartment together; intensity alone does not establish specificity (general IHC practice). |
| Nuclear staining accompanies cytoplasmic staining in otherwise plausible cells. | Nuclear signal is biologically plausible because SRPK1 shuttles and is reported in the nucleoplasm and nuclear speckles (UniProt Q96SB4: subcellular location). HPA tissue IHC describes a predominantly cytoplasmic pattern, so interpret a nuclear-only result cautiously (HPA: tissue IHC profile). |
| Staining is confined to an unexpected compartment, such as extracellular material or gland lumina. | Recheck its specificity and section morphology (general IHC practice). UniProt lists intracellular locations and no transmembrane segment or signal peptide; HPA tissue IHC reports cytoplasmic expression (UniProt Q96SB4: location and topology; HPA: tissue IHC profile). |
| The strongest signal is in a cell population reported as low, while nearby reported high cells are unstained. | Possible explanations include cross-reactivity, endogenous chromogenic detection activity, or a technical failure; the pattern alone cannot distinguish them (general IHC practice). Compare cell identities with HPA levels before calling it SRPK1: cholangiocytes and cardiomyocytes are Low, while several glandular cell populations are High (HPA: tissue IHC). |
| Haze covers cells and stroma, or a known high-staining tissue has no interpretable signal. | Diffuse haze prevents a compartment call; a blank positive control prevents a reliable negative call (general IHC practice). The supplied HPA profile lists high-staining cell populations but no negative tissue control (HPA: tissue IHC positive and negative lists). |
💡Expected SRPK1 appearanceCall a plausible positive when intracellular staining is predominantly cytoplasmic in an HPA high-staining cell population, with nuclear signal possible; a tissue-wide haze or extracellular deposit is suspect (HPA: tissue IHC profile and High cells; UniProt Q96SB4: nuclear shuttling; general IHC practice).
| Tissue and cell identity | HPA reports low tissue specificity and ubiquitous cytoplasmic expression, with High staining in selected glandular, epithelial and neuronal cells (HPA: tissue IHC). Compare like cell types; a whole-section average can hide their different staining levels (general IHC practice). |
| Intracellular distribution | Cytoplasmic and nuclear staining are plausible because SRPK1 shuttles between them (UniProt Q96SB4: subcellular location). A predominantly cytoplasmic tissue pattern is the HPA IHC expectation; nuclear intensity need not match it in every cell (HPA: tissue IHC profile; UniProt Q96SB4: nuclear shuttling). |
| Isoforms and tissue context | UniProt reports isoform 2 predominantly in testis and at lower levels in several other tissues; isoform 1 is reported only in testis (UniProt Q96SB4: tissue specificity). Those expression notes do not identify which isoform an IHC antibody detects; its epitope is unspecified here (UniProt Q96SB4: isoforms; supplied antibody data). |
| IHC evidence and interpretation limit | HPA016431 is Approved for IHC; the tissue profile has medium agreement between staining and RNA expression (HPA: antibody validation and tissue IHC reliability). These labels support using the reported pattern as a reference, while discordant staining still merits control review (general IHC practice). |
| IF/ICC Q: What pattern is expected? | A: Mainly nucleoplasm and cytosol, with additional plasma-membrane localization reported (HPA: subcellular ICC-IF, supported main locations and approved additional location). This ICC-IF observation informs compartment interpretation; it does not establish a paraffin-section IHC protocol (HPA: subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in colon glandular cells or bronchial respiratory epithelial cells. | These are reported High populations, so an empty result may reflect a failed IHC run; it does not establish absence of SRPK1 (HPA: tissue IHC; general IHC practice). | Check tissue preservation and a positive control, then review the catalog antibody's IHC-P retrieval, dilution and detection conditions (general IHC practice). No SRPK1-specific retrieval requirement is supplied. |
| Most of the section is uniformly brown, obscuring cell boundaries. | Excess chromogen, overly strong antibody signal, or endogenous detection activity can produce diffuse background (general IHC practice). | Compare a no-primary control, assess endogenous activity blocking, and adjust antibody or detection conditions using the validated IHC workflow (general IHC practice). |
| Only nuclei stain in tissue that otherwise resembles a reported High population. | Nuclear SRPK1 is possible, but HPA describes tissue staining as ubiquitously cytoplasmic (UniProt Q96SB4: nuclear shuttling; HPA: tissue IHC profile). | Check cytoplasmic visibility, counterstain and controls; report the compartment explicitly and avoid equating nuclear-only staining with the usual tissue pattern (general IHC practice; HPA: tissue IHC profile). |
| Low-reported cells stain more strongly than adjacent High-reported cells. | Cross-reactivity or endogenous detection activity is possible; cell identity or assay conditions could also explain the mismatch (HPA: tissue IHC levels; general IHC practice). | Verify the cell types and compare no-primary and positive controls before scoring SRPK1 expression (general IHC practice). HPA lists Low cholangiocytes and High glandular cells in several tissues (HPA: tissue IHC). |
| Extracellular or luminal deposits dominate the apparent signal. | That distribution conflicts with the reported intracellular locations and cytoplasmic tissue profile (UniProt Q96SB4: subcellular location; HPA: tissue IHC profile). | Inspect section morphology and controls for trapped chromogen or nonspecific deposits; score cellular staining separately (general IHC practice). |
| Staining differs between sections despite the same cell type being assessed. | Variation in section handling, antigen retrieval or detection can affect general IHC comparability; no target-specific SRPK1 fixation sensitivity is supplied (general IHC practice; supplied UniProt and HPA evidence). | Compare matched controls and recorded processing conditions, then interpret intensity only across comparable runs (general IHC practice). |
Experiment evidence
IHC & IF images showing SRPK1 staining
A01937-1 · IHCImmunohistochemistry of SRPK1 in human testis tissue with SRPK1 antibody at 5 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000096063-SRPK1/tissue/cerebral+cortex#imid_4153920
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000096063-SRPK1/tissue/colon#imid_4153862
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000096063-SRPK1/tissue/liver#imid_4153847
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000096063-SRPK1/tissue/kidney#imid_4153848
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA056486; green = SRPK1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA056486; green = SRPK1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SRPK1 IHC & IF
🧪Run appendix first and look for SRPK1 staining in glandular cells (HPA: High in appendix glandular cells). HPA detects SRPK1 in all 45 scored tissues, so there is no documented negative tissue; use no-primary and isotype controls, and expect empty glandular lumina and extracellular spaces on the positive slide to lack specific stain (HPA: no negative rows; standard IHC practice).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: None in HPA: SRPK1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SRPK1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and immunoglobulin class; for a polyclonal primary, use nonimmune IgG from the same host (standard IHC practice). Where available, compare with SRPK1 knockout material or perform a validated immunizing-peptide block, and quench endogenous peroxidase before chromogenic detection in appendix sections (standard IHC practice).
⚠️Feasibility: No supplied source reports an SRPK1-specific fixation window or fixation effect; the A01937-1 testis IHC caption gives 5 µg/ml but leaves the fixative unreported (selected IHC caption). Retrieval dependence is unreported, so optimize antigen retrieval on paraffin sections; the supplied evidence does not establish whether frozen sections or IF/ICC are easier, although ICC-IF images exist for A-431, U-251MG and U2OS (HPA subcellular). In appendix, score intracellular glandular-cell signal rather than chromogen deposited in glandular lumina (HPA: High in appendix glandular cells; standard IHC practice).
HPA tissue IHC evidence for SRPK1
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 → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: SRPK1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced SRPK1 IHC Tips
Troubleshoot SRPK1 chromogenic IHC in paraffin sections by checking retrieval, compartment-specific staining, controls and cell-level scoring.
Which retrieval conditions should I try first for weak SRPK1 staining?
Start with Tris-EDTA pH 9.0 heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). Allow sections to cool in the retrieval buffer, then compare staining with a matched section processed in the same run (standard IHC practice). The catalog antibody’s human testis image used 5 µg/ml, but its caption does not report retrieval conditions, so that image cannot establish this buffer’s performance (selected-SKU tissue-IHC caption). If staining remains weak, compare a milder retrieval condition on adjacent sections while keeping antibody concentration, detection and exposure to chromogen constant (standard IHC practice).
How should I assess whether fixation is masking SRPK1?
Target-specific fixation sensitivity is unknown because the selected-SKU tissue-IHC caption does not state a fixative (selected-SKU tissue-IHC caption). Record the specimen’s fixative and fixation time, then compare sections from similarly handled blocks before attributing weak staining to fixation (standard IHC practice). Use the page’s Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min as the starting condition for each comparison (page retrieval setting). If comparable blocks differ, review fixation history alongside section age, retrieval and detection controls; neither SRPK1 localisation nor its phosphorylation sites establish a fixation effect (UniProt Q96SB4; standard IHC practice).
Should SRPK1 staining be cytoplasmic, nuclear, or both?
Assess cytoplasmic and nuclear signal separately: tissue IHC shows ubiquitous cytoplasmic expression, while subcellular data support cytosol and nucleoplasm localisation (HPA tissue IHC; HPA subcellular). SRPK1 shuttles between cytoplasm and nucleus, and stress or altered Hsp90 activity can induce nuclear translocation (UniProt Q96SB4 localisation). In chromogenic sections, use a nuclear counterstain to decide whether brown signal lies within nuclei, around them, or across both compartments (standard IHC practice). Compare like cell populations within the same section, because a changed nuclear-to-cytoplasmic pattern need not represent a change in total SRPK1 abundance (UniProt Q96SB4 localisation; standard IHC interpretation).
Can this stain distinguish SRPK1 isoforms or phosphorylation states?
Do not assign an isoform from staining alone: SRPK1 has 3 annotated isoforms, and the supplied tissue image does not identify the antibody epitope (UniProt Q96SB4 isoforms; selected-SKU tissue-IHC caption). Isoform 2 predominates in testis, whereas isoform 1 is reported there at lower levels (UniProt Q96SB4 tissue specificity). SRPK1 also has annotated phosphoserines, including positions 309, 311 and 333, but the caption does not establish phospho-specific recognition (UniProt Q96SB4 modified residues; selected-SKU tissue-IHC caption). To make an isoform or modification claim, first establish epitope coverage and validate selectivity with appropriate reference material (standard IHC validation practice).
How can IF help resolve ambiguous SRPK1 compartments in tissue?
Use this secondary IF experiment to compare SRPK1 with a marker of the cell population being evaluated, such as glandular cells in colon sections (HPA tissue IHC: high in colon glandular cells). Choose a fluorophore channel that separates the antibody signal from the specimen’s measured autofluorescence, and include single-stain controls for multiplexing (standard IF practice). Because the supported SRPK1 locations include cytosol and nucleoplasm, test gentle permeabilisation that permits access to intracellular epitopes after fixation (HPA subcellular; standard IF practice). Compare nuclear and cytoplasmic signal within marker-positive cells, while treating the catalog antibody’s tissue image as IHC evidence rather than IF validation (selected-SKU tissue-IHC caption).
What should I check when brown signal obscures SRPK1 staining?
Inspect a section without primary antibody to gauge detection-system background before changing the SRPK1 antibody concentration (standard chromogenic IHC practice). Block endogenous peroxidase for an HRP-based workflow, and check whether the DAB reaction was developed long enough to obscure cellular boundaries (standard chromogenic IHC practice). Compare the test section with the selected-SKU human testis image, whose stated antibody concentration is 5 µg/ml; its caption does not specify fixation or retrieval (selected-SKU tissue-IHC caption). Titrate antibody and development time on adjacent sections, and retain a consistent counterstain so diffuse brown deposits can be distinguished from cytoplasmic or nuclear signal (standard IHC practice; HPA subcellular).
How should I score SRPK1 across specimens with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since tissue IHC describes widespread cytoplasmic expression and subcellular data also support nucleoplasmic signal (HPA tissue IHC; HPA subcellular). For intensity, calculate an H-score from the percentages of target cells at each staining grade; for prevalence, report the percentage of positive target cells (standard IHC scoring practice). Normalise each measure to the number of evaluable cells in the specified population, or report positive-cell density per mm² of evaluable tissue when cell counting is impractical (standard IHC scoring practice). Keep retrieval, detection, counterstain and scoring thresholds consistent across specimens, and record nuclear and cytoplasmic results separately (standard IHC practice).
How do I separate convincing SRPK1 staining from artefact?
Convincing staining should resolve within intact cells and be assessed against the supported cytoplasmic pattern and possible nucleoplasmic localisation (HPA tissue IHC; HPA subcellular). Check whether the signal belongs to the intended cell population; high staining is reported in colon glandular cells, while low staining is reported in liver cholangiocytes (HPA tissue IHC). Treat staining confined to section edges, necrotic areas or primary-antibody-omitted controls as suspect, and investigate endogenous enzyme activity in an HRP–DAB workflow (standard chromogenic IHC practice). A nuclear shift alone can be biologically plausible for shuttling SRPK1, but require reproducible cell-level localisation and appropriate controls before interpreting it as translocation (UniProt Q96SB4 localisation; standard IHC practice).
Boster reagents
Best SRPK1 / SRSF protein kinase 1 IHC Antibodies
A01937-1 has real IHC data in human testis (image caption: human testis). It lists human, mouse and rat reactivity (catalog: reactivity), with IHC validation reported in human samples (datasheet: validation).
Real IHC data
Anti-SRSF protein kinase 1 SRPK1 Antibody
Cat # A01937-1
A01937-1 is listed for IHC-P (catalog: applications) and shows human testis staining at 5 μg/ml (image caption: human testis, 5 μg/ml). It lists human, mouse and rat reactivity (catalog: reactivity), while the reported IHC validation is in human samples (datasheet: validation).
Which to pick: Choose A01937-1 for paraffin-section IHC (catalog: IHC-P); its image shows human testis staining at 5 μg/ml (image caption: human testis, 5 μg/ml), and the fixative is unreported (image caption: no fixative specified). No SKU here lists IF/ICC or has an IF image (catalog: applications and image alts). For mouse or rat tissue, A01937-1 lists reactivity with both species (catalog: reactivity), but its reported IHC validation is in human samples (datasheet: validation).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96SB4 (SRPK1_HUMAN, SRSF protein kinase 1).
- Human Protein Atlas. SRPK1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SRPK1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and cytosol. In addition localized to the plasma membrane..
- Human Protein Atlas. SRPK1 antibody validation summary (2 antibodies).
- Serine-arginine protein kinase 1 (SRPK1) inhibition as a potential novel targeted therapeutic strategy in prostate cancer. Oncogene 2015 — PMC4351909.
- SRPK1 Dissimilarly Impacts on the Growth, Metastasis, Chemosensitivity and Angiogenesis of Glioma in Normoxic and Hypoxic Conditions. Journal of Cancer 2013 — PMC3842442.
- Inhibition of SRPK1, a key splicing regulator, exhibits antitumor and chemotherapeutic-sensitizing effects on extranodal NK/T-cell lymphoma cells. BMC cancer 2022 — PMC9609466.
- SRPK1 Promotes Glioma Proliferation, Migration, and Invasion through Activation of Wnt/β-Catenin and JAK-2/STAT-3 Signaling Pathways. Biomedicines 2024 — PMC10886746.
- PubMed PMID:8208298 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:11509566 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SRPK1 (SRSF protein kinase 1). Expected localisation Predominantly cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A01937-1 at 5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. 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 A01937-1); 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 Tris-EDTA buffer, pH 9.0 , 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-SRPK1, 5 μg/ml (datasheet A01937-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01937-1); Caveat: Stress or Hsp90 inhibition can shift SRPK1 into nuclei (UniProt); Isoform / epitope: 3 isoforms; verify epitope coverage across variants (UniProt); Regulation: Isoform 2 predominates in testis (UniProt). Evidence: UniProt Q96SB4, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and cytosol. In addition…, Datasheet A01937-1, PMC4351909, PMC3842442. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SRPK1/immunohistochemistry-ihc-guide.