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SRSF7 / Serine/arginine-rich splicing factor 7 · IHC design guide
Design Immunohistochemistry for SRSF7
This guide maps SRSF7 nuclear staining across most tissues (HPA tissue IHC). It pairs cell-level examples with the 1:100–1:300 IHC dilution range (datasheet A30768) for paraffin sections.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SRSF7 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 | Nuclear staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining; high in alveolar type I cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A30768) | |
| Positive control | Lung+4 more · see all | |
| Negative control | Adipose tissue+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes have no detectable staining (HPA tissue IHC) | |
| Regulation | Staining intensity varies by cell type (HPA tissue IHC) | |
| Isoform / epitope | Four isoforms; verify antibody epitope coverage (UniProt) |
Section 1
Recommended SRSF7 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A30768) is followed by published human tissue protocols (PMC5844074; PMC12249523).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A30768) |
| Fixation | Image fixative and duration unreported (datasheet A30768); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A30768); 20 min, 95–100 °C (standard) |
| 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-SRSF7, 1:100 - 1:300 (datasheet A30768) |
| 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 | SRSF7-positive staining in alveolar cells type I of lung (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 9.0 (datasheet A30768); neither published excerpt specifies retrieval conditions (PMC5844074 methods; PMC12249523 methods).
Section 2
What Is the Expected SRSF7 Staining Pattern?
SRSF7 is expected mainly in nuclei across most tissues (HPA tissue IHC: nuclear expression in most tissues). Strong examples are lung alveolar type I cells, skin granular-layer cells, and round or early spermatids (HPA tissue IHC: High). UniProt also lists cytoplasm, so a faint cytoplasmic component is possible (UniProt Q16629: nucleus; cytoplasm). The HPA tissue assessment is Supported, with medium consistency between staining and RNA data (HPA tissue IHC: reliability).
| Distinct nuclear staining in alveolar type I cells, granular-layer cells, or round or early spermatids. | This matches the reported compartment and High cell-level staining (HPA tissue IHC: nuclear profile; High in these cells). Judge intensity against neighboring cells and the negative control on the same slide (general IHC practice). A High designation describes HPA observations; it does not guarantee equally strong staining with every antibody or run. |
| Predominantly cytoplasmic staining with little nuclear signal in a tissue expected to stain. | The compartment balance conflicts with the HPA nuclear IHC profile and enhanced nucleoplasmic ICC-IF location (HPA tissue IHC; HPA subcellular). Cytoplasmic localization is also listed (UniProt Q16629: subcellular location), so cytoplasm alone does not prove an artefact. Review controls and antibody specificity before interpreting it as SRSF7. |
| Strong signal in adipocytes, chondrocytes, or splenic red-pulp cells. | These cells are reported as Not detected in the sampled tissues (HPA tissue IHC: adipose tissue, soft tissue, spleen). Unexpected staining could reflect cross-reactivity or endogenous chromogenic activity (general IHC practice). Compare nuclear localization, neighboring expected-positive cells, and a control omitting the primary antibody before assigning a cause. |
| Diffuse color over nuclei, cytoplasm, and tissue spaces, with no clear cell boundaries. | A widespread deposit without a nuclear pattern is difficult to score as specific SRSF7 (HPA tissue IHC: nuclear profile). Excess detection background or inadequate blocking are possible explanations (general IHC practice). A primary-omission control helps distinguish detection-related signal from staining that depends on the primary antibody. |
| No nuclear signal in an otherwise interpretable lung, skin, or testis section. | First confirm that the section contains the reported positive cell type: alveolar type I cells, granular-layer cells, or round or early spermatids (HPA tissue IHC: High). Missing those cells changes the comparison. If present, a failed stain is possible; the HPA Supported rating and medium RNA-staining consistency do not make every specimen an assured positive (HPA tissue IHC: reliability). |
💡Expected SRSF7 appearanceCall a result positive when staining is chiefly nuclear in the reported cell type, strongest in HPA High examples; broad background or strong signal in HPA Not detected cells warrants control-based review (HPA tissue IHC: profile and levels; general IHC practice).
| Compartment and cell choice | Expect nuclear staining in most tissues, with HPA High examples in lung, skin, and testis (HPA tissue IHC). UniProt also lists cytoplasm (UniProt Q16629: location); interpret its significance alongside nuclear staining and controls. |
| Strength across tissues | HPA reports Medium staining in adrenal and appendix glandular cells, bone-marrow hematopoietic cells, and bronchial respiratory epithelial cells (HPA tissue IHC). These provide context when comparing fields; the HPA assessment is Supported with medium RNA-staining consistency (HPA tissue IHC: reliability). |
| Isoforms and epitope coverage | Four SRSF7 isoforms are listed (UniProt Q16629: isoforms 1–4). Without an antibody epitope in the supplied evidence, coverage of individual isoforms cannot be assigned. Interpret differing intensity cautiously rather than attributing it to a specific isoform. |
| Processing and modifications | UniProt lists a single 1–238 chain, no signal peptide or propeptide, and several modified residues (UniProt Q16629: processing; modified residues). These facts do not predict staining intensity or an antigen-retrieval setting. Target-specific fixation sensitivity is unreported in the supplied evidence. |
| Chromogenic detection controls | Endogenous tissue activity can resemble chromogen signal, and nonspecific detection can add background (general IHC practice). Use an appropriate primary-omission control and the detection system’s recommended activity controls when an unexpected cell type stains; these checks do not themselves establish target specificity. |
| IF/ICC Q&A: What compartment should IF show? | A nucleoplasmic signal is the reference pattern (HPA subcellular: enhanced nucleoplasm). HPA lists A-431, U-251MG, and U2OS ICC-IF images; both listed antibodies have Enhanced ICC validation (HPA subcellular; HPA antibodies). IF/ICC staining conditions belong to its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive nuclei remain unstained. | The reported positive cells may be absent, or the IHC run may have failed (HPA tissue IHC: High cell types; general IHC practice). | Verify the cell type in the section, then check an independent positive control, detection reagents, and the antibody’s validated IHC-P conditions (general IHC practice). Do not infer target-specific fixation sensitivity from this result. |
| Staining is mainly cytoplasmic. | A dominant cytoplasmic pattern differs from HPA nuclear IHC and nucleoplasmic ICC-IF, although UniProt lists cytoplasm (HPA tissue IHC; HPA subcellular; UniProt Q16629). | Check whether expected-positive nuclei stain on the same run. Compare primary-omission and detection controls, then reassess localization before scoring cytoplasmic color as specific (general IHC practice). |
| Adipocytes, chondrocytes, or red-pulp cells stain strongly. | Those cell types were Not detected in the HPA tissue observations; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Inspect the signal’s compartment and run a primary-omission control. If signal persists without primary antibody, review detection-related background; if it depends on primary antibody, investigate specificity (general IHC practice). |
| A diffuse haze obscures nuclear detail. | Excess background can obscure the expected nuclear pattern (HPA tissue IHC: nuclear profile; general IHC practice). | Review blocking, washing, antibody concentration, and chromogen development against the established IHC-P procedure; compare a primary-omission control (general IHC practice). Avoid treating a change in background as evidence of a target-specific fixation effect. |
| Signal appears only in scattered cells within a presumed positive tissue. | HPA levels refer to named cell populations, and section composition can vary (HPA tissue IHC: cell-level observations). | Locate the relevant cells before judging sensitivity: alveolar type I cells in lung, granular-layer cells in skin, or round or early spermatids in testis (HPA tissue IHC: High). Score the appropriate population rather than the entire section. |
| IF/ICC appears inconsistent with the IHC slide. | HPA’s ICC-IF reference is nucleoplasmic, while its tissue IHC summary says nuclear expression in most tissues (HPA subcellular; HPA tissue IHC). | Compare localization within each method’s own controls and cell context (general IHC/IF practice). Use the separate IF/ICC guide for its conditions; this section supplies no IF protocol option. |
Experiment evidence
IHC & IF images showing SRSF7 staining
A30768 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Tris-EDTA,pH9.0 was used for antigen retrieval. 2 Antibody was diluted at
A05827 · IHCImmunohistochemistry (IHC) analyzes of SRSF7 (N103) pAb in paraffin-embedded human breast carcinoma tissue at 1:50.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115875-SRSF7/tissue/cerebral+cortex#imid_18173535
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115875-SRSF7/tissue/colon#imid_18173453
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115875-SRSF7/tissue/liver#imid_18173446
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000115875-SRSF7/tissue/kidney#imid_18173463
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043850; green = SRSF7, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043850; green = SRSF7, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SRSF7 IHC & IF
🧪Run lung first: alveolar type I cells should show SRSF7 staining (HPA: High in alveolar cells type I). Use adipose tissue as a negative comparator, focusing on adipocytes (HPA: Not detected in adipocytes); on the lung slide, cells without specific nuclear staining provide a background check, but HPA does not identify a validated internal negative cell type (HPA: High in alveolar cells type I).
Positive control tissue: Lung (Alveolar cells type I, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SRSF7 in A-431, U-251MG, U2OS, U2OS, siRNA 1 (10x), U2OS, siRNA 1 (40x), U2OS, siRNA 2 (10x), U2OS, siRNA 2 (40x), U2OS, scrambled (10x), U2OS, scrambled (40x), with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species and clonality; and a biological specificity control using target knockout material or a cognate peptide block, if available (standard IHC practice). For chromogenic lung IHC, block endogenous peroxidase and inspect pigment-associated background; for IF, assess tissue autofluorescence (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A30768 paraffin-section caption does not state its fixative (A30768 caption: fixative unreported). The caption reports Tris-EDTA retrieval at pH 9.0 for paraffin-embedded tonsil, so retrieval conditions should be checked in lung; it does not establish a requirement for that retrieval condition in lung (A30768 caption: tonsil retrieval). Paraffin IHC has a supplied protocol example, while frozen-section performance and whether IF is easier are unreported; lung pigment can complicate chromogenic interpretation (A30768 caption: paraffin-embedded tonsil; standard IHC practice).
HPA tissue IHC evidence for SRSF7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Lung | Alveolar cells type I | High | Protein (IHC) | HPA → |
| Skin | Cells in granular layer | High | Protein (IHC) | HPA → |
| Testis | Round or early spermatids | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced SRSF7 IHC Tips
Troubleshoot SRSF7 staining by checking retrieval, nuclear signal, background and scoring against the available tissue evidence.
Which retrieval conditions should I try first for SRSF7 in paraffin sections?
Start with heat-mediated retrieval in Tris-EDTA, pH 9.0 (datasheet A30768). The paraffin-section tonsil image used that buffer with primary antibody at 1:200 overnight at 4°C, but does not establish that those conditions work equally well in every tissue (caption A30768). If nuclear staining is weak, vary heating duration and cooling under otherwise identical conditions, then assess tissue integrity and signal together (standard IHC practice). Compare with a no-primary control and a consistently processed positive reference, because nonspecific chromogen deposits can make increased retrieval appear successful (standard IHC practice; HPA: nuclear expression in most tissues).
How can I assess whether fixation is limiting SRSF7 staining?
Target-specific fixation sensitivity for SRSF7 is unknown from the supplied evidence, and the selected paraffin-section caption does not report a fixative (caption A30768). Record the fixative, fixation duration, section age and processing history for each specimen before comparing staining (standard IHC practice). If nuclear signal differs between batches, stain comparable sections together using the same retrieval, 1:200 primary dilution and detection conditions where applicable (caption A30768; standard IHC practice). Check morphology and a processed positive reference before attributing a weak result to fixation; fixation effects cannot be inferred from the tissue staining profile or SRSF7’s modifications (standard IHC practice; HPA: tissue IHC; UniProt Q16629).
Where should SRSF7 staining appear, and how should I handle cytoplasmic signal?
Expect predominantly nuclear staining in paraffin sections: the tissue profile reports nuclear expression in most tissues, and subcellular imaging places SRSF7 in the nucleoplasm (HPA: tissue IHC; HPA: nucleoplasm). UniProt also lists cytoplasm, so cytoplasmic signal deserves assessment rather than automatic rejection (UniProt Q16629: subcellular location). Compare nuclear and cytoplasmic staining within intact cells, and check whether the cytoplasmic pattern persists when primary antibody is omitted (standard IHC practice). Score the compartments separately, especially in mixed tissue, and avoid treating diffuse cytoplasmic chromogen as confirmation of SRSF7 without a clean nuclear reference (standard IHC practice; HPA: nuclear expression in most tissues).
Could isoforms or modifications explain different SRSF7 staining patterns?
SRSF7 has 4 listed isoforms, an RNA recognition motif at residues 11–84, and reported phosphorylation at residues 32, 163, 165 and 167 (UniProt Q16629). Those features make epitope accessibility a reasonable question, but the supplied material does not map this antibody’s epitope or establish isoform selectivity (UniProt Q16629; caption A30768). Check the antibody’s documented immunogen and validation before interpreting differences between specimens as isoform changes (standard IHC practice). Keep retrieval and primary incubation constant across the comparison; orthogonal evidence is needed to assign a particular isoform or modification to a chromogenic staining pattern (standard IHC practice).
How should I assess SRSF7 by IF alongside a cell-type marker?
Use the separate IF/ICC guide to develop that assay; the 1:200 dilution and pH 9.0 retrieval come from a paraffin-section IHC caption (caption A30768). For multiplexing, pair SRSF7 with a validated marker for the cell type under study and check that their fluorescence channels remain distinct (standard IF practice). In tissue with autofluorescence, compare unstained controls across channels and consider a far-red fluorophore for the weaker signal (standard IF practice). Because SRSF7 is nucleoplasmic and has no transmembrane segment, choose permeabilisation that allows antibody access through the cell and nuclear boundaries, then verify preserved nuclear morphology (HPA: nucleoplasm; UniProt Q16629: topology; standard IF practice).
What should I change when chromogenic background obscures SRSF7 nuclei?
First compare the stained section with a no-primary control to locate background from the detection system or tissue itself (standard IHC practice). The selected tonsil caption reports primary and secondary antibody dilutions of 1:200, with secondary incubation for 45 minutes at room temperature (caption A30768). Check peroxidase blocking and the duration of chromogen development, and titrate antibody concentration if diffuse staining persists (standard chromogenic IHC practice). Review edges, folds and damaged regions separately from intact nuclei; the expected nuclear tissue pattern makes uniform extranuclear deposits a poor basis for calling SRSF7 positive (HPA: nuclear expression in most tissues; standard IHC practice).
What is a defensible way to quantify SRSF7 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define regions and cell classes before scoring, then report the percentage of positive nuclei and a consistent nuclear intensity scale (standard IHC practice; HPA: nuclear expression in most tissues). If an H-score is useful, multiply the percentage at each intensity category by its category value and sum the results on a 0–300 scale (standard IHC practice). Normalise counts to the number of evaluable nuclei, or report positive-cell density per mm² with the sampled tissue area stated (standard IHC practice). Keep section processing, imaging and thresholds consistent, and record cytoplasmic signal separately so it does not inflate the nuclear result (standard IHC practice; UniProt Q16629: cytoplasmic location).
How can I distinguish convincing SRSF7 staining from tissue artefact?
Give greatest weight to staining in intact nuclei: SRSF7 is predominantly nuclear in tissue and nucleoplasmic in subcellular imaging (HPA: tissue IHC; HPA: nucleoplasm). Check the annotated cell population before interpreting a negative field; HPA reports high staining in lung alveolar type I cells and no detection in adipocytes (HPA: lung; HPA: adipose tissue). Exclude section edges, folds and necrotic areas from interpretation, and use a no-primary control to assess endogenous enzyme activity or detection background (standard IHC practice). A compartment mismatch or control staining calls for further validation, while cytoplasmic signal alone does not establish a specific SRSF7 result (UniProt Q16629: cytoplasmic location; standard IHC practice).
Boster reagents
Best SRSF7 / Serine/arginine-rich splicing factor 7 IHC Antibodies
Two catalog antibodies have IHC images from paraffin-embedded human tonsil and breast carcinoma (image captions); listed reactivity covers human and mouse for both, plus rat for A05827 (catalog).
Real IHC data
Anti-SFRS7 Antibody
Cat # A30768
Real IHC data
Anti-SRSF7 (N103) Antibody
Cat # A05827
A30768 lists IHC, IF and ICC applications for human and mouse, with an IHC image from paraffin-embedded human tonsil (catalog; image caption). A05827 lists IHC for human, mouse and rat, with an IHC image from paraffin-embedded human breast carcinoma (catalog; image caption).
Which to pick: For tissue IHC, choose A30768 for its documented paraffin-section retrieval and 1:200 staining example, or A05827 for its paraffin-section breast carcinoma example at 1:50 (respective IHC image captions). For IF/ICC, choose A30768 because those applications are listed for this rabbit polyclonal antibody; no IF image is supplied (catalog). For rat-reactive IHC, choose the rabbit polyclonal A05827, which lists rat reactivity; neither IHC caption reports the fixative (catalog; respective IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q16629 (SRSF7_HUMAN, Serine/arginine-rich splicing factor 7).
- Human Protein Atlas. SRSF7 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. SRSF7 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. SRSF7 antibody validation summary (2 antibodies).
- SRSF7 knockdown promotes apoptosis of colon and lung cancer cells. Oncology letters 2018 — PMC5844074.
- Serine/Arginine-Rich Splicing Factor 7 Knockdown Inhibits Aerobic Glycolysis and Growth in HepG2 Cells by Regulating PKM2 Expression. Current issues in molecular biology 2024 — PMC11119032.
- hnRNP A1-mediated translational regulation of the G quadruplex-containing RON receptor tyrosine kinase mRNA linked to tumor progression. Oncotarget 2016 — PMC4941351.
- Multi-Omics Integration: Predicting Progression and Optimizing Clinical Treatment of Hepatocellular Carcinoma Through Malignant-Cell-Related Genes. International journal of molecular sciences 2025 — PMC12249523.
- PubMed PMID:8013463 — UniProt-cited evidence.
- PubMed PMID:7629084 — UniProt-cited evidence.
- PubMed PMID:20857263 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SRSF7 (Serine/arginine-rich splicing factor 7). Expected localisation Nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A30768). Validated IHC antibody: A30768 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Lung (Alveolar cells type I); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A30768); 2. Fixation: Image fixative and duration unreported (datasheet A30768); 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: Tris-EDTA pH 9.0 (datasheet A30768); 20 min, 95–100 °C (standard); 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-SRSF7, 1:100 - 1:300 (datasheet A30768); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Adipocytes have no detectable staining (HPA tissue IHC); Isoform / epitope: Four isoforms; verify antibody epitope coverage (UniProt); Regulation: Staining intensity varies by cell type (HPA tissue IHC). Evidence: UniProt Q16629, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A30768, PMC5844074, PMC12249523. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SRSF7/immunohistochemistry-ihc-guide.