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SSRP1 / FACT complex subunit SSRP1 · IHC design guide
Design Immunohistochemistry for SSRP1
Plan chromogenic IHC on paraffin sections around the expected nuclear staining pattern (HPA tissue IHC). Use high-staining parathyroid glandular cells as a positive reference, and interpret results with the reported low agreement between staining and RNA data in mind (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SSRP1 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 nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in most tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02606-1) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin tissue fixation consistent across samples and controls. (selected-SKU IHC image A02606-1) | |
| Caveat | Low agreement between staining and RNA data (HPA tissue IHC) | |
| Regulation | Recruited to actively transcribed loci (UniProt) | |
| Isoform / epitope | No isoforms annotated; chain spans aa 2–709 (UniProt) |
Section 1
Recommended SSRP1 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet: A02606-1). The four published IHC protocols below provide tissue specific conditions (PMC5780019; PMC5422336; PMC6909804; PMC3248156).
| Sample | Paraffin-embedded human appendix adenocarcinoma tissue; fixative not specified (datasheet A02606-1) |
| Fixation | Image fixative and duration unreported (datasheet A02606-1); 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 A02606-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02606-1) |
| Primary antibody | Rabbit anti-SSRP1, 2-5 μg/ml (datasheet A02606-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02606-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02606-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SSRP1-positive staining in purkinje cells of cerebellum (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet: A02606-1); citrate pH 6.0 is a published alternative for other antibodies (PMC5780019; PMC3248156).
Section 2
What Is the Expected SSRP1 Staining Pattern?
SSRP1 is predominantly nuclear, with nucleolar localization also reported (UniProt Q08945; HPA: nucleoplasm and nucleoli). In paraffin sections, expect nuclear staining across many tissues, including strong staining in Purkinje cells, parathyroid glandular cells, placental trophoblastic cells and cells in seminiferous ducts (HPA: tissue IHC). Interpret intensity cautiously: HPA rates the tissue IHC pattern Approved but reports low consistency between antibody staining and RNA expression data (HPA: reliability). SSRP1 has no transmembrane segment (UniProt Q08945 topology).
| Nuclear chromogen in Purkinje cells or placental trophoblastic cells. | This fits two high-staining cell populations in the HPA tissue IHC record (HPA: High in Purkinje cells; High in trophoblastic cells). Score the identified cells and their nuclear signal, since an adjacent cell population need not share the same intensity (HPA: cell-specific tissue IHC observations). |
| Predominantly cytoplasmic or membrane staining, with little nuclear signal. | This conflicts with the reported nuclear tissue pattern and SSRP1's nuclear and chromatin localization (HPA: nuclear expression in most tissues; UniProt Q08945 localization). Review morphology, controls and detection background before interpreting the compartment as SSRP1 (standard IHC practice). |
| Strong signal in a cell population listed as not detected, such as adipocytes. | HPA reports adipocytes as not detected (HPA: adipose tissue). Check cell identity and staining controls; cross-reactivity or endogenous detection activity are possible explanations, not established diagnoses (standard IHC practice). An HPA negative observation alone does not prove every specimen will be negative. |
| Broad haze or chromogen over both nuclei and surrounding tissue. | Diffuse staining obscures the nuclear pattern expected for SSRP1 (HPA: nuclear tissue expression; UniProt Q08945 localization). Review background in the negative control and detection reagents before scoring cells (standard IHC practice). |
| No nuclear signal in an adequately represented known-positive tissue. | Absence is unexpected in, for example, parathyroid glandular cells, which HPA scores High (HPA: parathyroid gland). Confirm that the expected cells are present, then review the IHC-validated antibody, retrieval and detection controls (standard IHC practice). |
💡Expected SSRP1 appearanceCall a result positive when the identified cells show predominantly nuclear chromogen, especially a strong signal in an HPA High population; diffuse cytoplasmic or membrane-only staining is suspect (HPA: nuclear expression and cell-specific levels; UniProt Q08945 localization).
| Cell population and tissue | Expected intensity depends on the cells examined: HPA lists Purkinje cells as High, skin keratinocytes as Medium and stomach glandular cells as Low (HPA: tissue IHC). Compare like cell populations when judging a section; a weak stomach result need not match a Purkinje-cell result. |
| Subnuclear localization | Nuclear staining is the main IHC expectation (HPA: tissue IHC). Nucleolar localization is also reported by UniProt and supported in HPA ICC-IF; HPA lists centrosome localization as uncertain in ICC-IF (UniProt Q08945 localization; HPA: subcellular). Do not require a centrosomal IHC signal. |
| Antibody evidence | HPA lists HPA002697 as IHC Approved and HPA002696 as ICC Supported (HPA: antibody validation). The tissue profile is also labeled Approved with low staining-versus-RNA consistency (HPA: reliability); use cell identity and controls when interpreting an unexpected result. |
| Antigen retrieval and fixation | No target-specific retrieval condition or fixation sensitivity is supplied by UniProt or HPA. Select and assess retrieval through the IHC-validated antibody's instructions and section controls (standard IHC practice); neither HPA staining levels nor UniProt modifications establish a fixation effect. |
| Topology and processing | UniProt reports no transmembrane segment or signal peptide and lists a FACT-complex chain spanning residues 2–709 (UniProt Q08945 topology and processing). These annotations support a nuclear interpretation; they do not identify the antibody epitope or establish a shedding effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| A high-expected control shows no nuclear stain. | The expected cells may be absent, or the IHC detection workflow may have failed; the image alone cannot distinguish these possibilities (HPA: High in parathyroid glandular cells; standard IHC practice). | Verify cell identity and section quality, then check the IHC-validated antibody's instructions, retrieval run and detection controls (standard IHC practice). |
| Only cytoplasm or cell outlines are stained. | The compartment differs from the reported nuclear pattern (HPA: nuclear expression in most tissues; UniProt Q08945 localization). Nonspecific binding or detection background is possible (standard IHC practice). | Compare a known-positive nuclear control with a negative detection control; reassess cell boundaries and staining specificity before scoring (standard IHC practice). |
| An HPA not-detected population stains strongly. | Cross-reactivity or endogenous detection activity is possible; HPA lists adipocytes and cardiomyocytes as not detected (HPA: tissue IHC; standard IHC practice). | Confirm the cell population and inspect negative controls. Treat the result as unresolved until nuclear localization and antibody specificity are supported (standard IHC practice). |
| Background obscures nuclear boundaries. | Diffuse chromogen can prevent compartment scoring regardless of the expected SSRP1 pattern (standard IHC practice; HPA: nuclear tissue expression). | Check negative controls and detection background; reassess blocking, antibody concentration and chromogen development using the IHC-validated antibody's instructions (standard IHC practice). |
| A weak nucleus in stomach seems inconsistent with a strong positive control. | HPA scores stomach glandular cells Low and Purkinje cells High (HPA: tissue IHC). Those cell populations have different observed staining levels. | Score each identified population against its own HPA expectation and the run controls; avoid applying one tissue's intensity threshold to another (HPA: tissue IHC; standard IHC practice). |
| Q: What should IF/ICC show? | HPA supports nucleoplasm and nucleoli in ICC-IF and labels centrosome localization uncertain (HPA: subcellular). UniProt also reports nucleus and nucleolus (UniProt Q08945 localization). | A: Look for predominantly nucleoplasmic fluorescence, with possible nucleolar signal; do not require centrosome staining (HPA: subcellular). Consult the separate IF/ICC guide for its workflow. |
Experiment evidence
IHC & IF images showing SSRP1 staining
A02606-1 · IHCIHC analysis of SSRP1 using anti-SSRP1 antibody (A02606-1). SSRP1 was detected in a paraffin-embedded section of human appendix adenocarcino
A02606-1 · IHCIHC analysis of SSRP1 using anti-SSRP1 antibody (A02606-1). SSRP1 was detected in a paraffin-embedded section of human breast cancer tissue.
A02606-1 · IFIF analysis of SSRP1 using anti-SSRP1 antibody (A02606-1) and anti-Tubulin Alpha antibody (M03989-3). SSRP1 was detected in immunocytochemic
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000149136-SSRP1/tissue/cerebral+cortex#imid_992504
HPA · StomachHuman Protein Atlas IHC, Stomach. Source: https://www.proteinatlas.org/ENSG00000149136-SSRP1/tissue/stomach+2#imid_992372
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000149136-SSRP1/tissue/colon#imid_992430
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000149136-SSRP1/tissue/liver#imid_992431
HPA · ICC-IF HAP1Human Protein Atlas immunofluorescence, HAP1, antibody HPA002696; green = SSRP1, blue = nucleus, red = microtubules.
HPA · ICC-IF HAP1Human Protein Atlas immunofluorescence, HAP1, antibody HPA002696; green = SSRP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SSRP1 IHC & IF
🧪Run parathyroid gland first: glandular cells should show nuclear SSRP1 staining (HPA: High in parathyroid glandular cells; UniProt Q08945: nuclear localization). Use heart muscle as the negative tissue and score cardiomyocytes (HPA: Not detected in cardiomyocytes); on the positive slide, anucleate erythrocytes, if present, should show no nuclear signal (standard IHC practice).
Positive control tissue: Cerebellum (Purkinje cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SSRP1 in HAP1, Rh30, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control, alongside an SSRP1 knockout control if available (selected-SKU caption: rabbit primary; standard IHC practice). Check the parathyroid section for endogenous peroxidase signal before interpreting DAB staining (standard chromogenic IHC practice).
⚠️Feasibility: The selected-SKU paraffin-section caption reports heat-mediated retrieval in EDTA at pH 8.0 and primary antibody at 2 μg/ml overnight at 4°C, but its fixative is unreported (selected-SKU caption). A target-specific fixation window, retrieval dependency, and any advantage of frozen sections or IF are unreported in the supplied evidence; use the documented IHC conditions as a starting point (selected-SKU caption). Assess any parathyroid background against the no-primary control before scoring nuclear staining (standard chromogenic IHC practice).
HPA tissue IHC evidence for SSRP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Skin | Keratinocytes | Medium | Protein (IHC) | HPA → |
| Stomach | Glandular cells | Low | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SSRP1 IHC Tips
Troubleshoot SSRP1 staining by checking retrieval, nuclear localization, assay controls, and cell-specific scoring before interpreting chromogenic signal.
What should I change when nuclear SSRP1 staining is weak after antigen retrieval?
Start with heat-mediated EDTA pH 8.0 retrieval for paraffin sections stained with A02606-1 (datasheet A02606-1). The selected tissue-IHC example used that retrieval before an overnight primary incubation at 4°C, but did not report the retrieval time or temperature (caption A02606-1). If nuclei remain weak, compare retrieval durations on adjacent sections while holding antibody concentration, detection, and development constant (standard IHC practice). Include a tissue region expected to show nuclear staining, such as appendix lymphoid tissue, alongside a no-primary control (HPA tissue IHC: medium in appendix lymphoid tissue; standard IHC practice). Excess retrieval can damage morphology or raise nonspecific staining, so judge signal together with nuclear detail (standard IHC practice).
Could fixation explain variable SSRP1 staining between paraffin blocks?
The selected A02606-1 tissue-IHC caption describes a paraffin section but does not state its fixative, so SSRP1-specific fixation sensitivity is unknown (caption A02606-1). Record each block’s fixative, fixation duration, and processing history before comparing staining intensities across specimens (standard IHC practice). For a controlled comparison, process adjacent samples with documented fixation conditions, then stain them together using EDTA pH 8.0 retrieval and the same detection run (datasheet A02606-1; standard IHC practice). Assess nuclear signal alongside preservation of nuclei and tissue architecture; damaged morphology can make a staining difference difficult to attribute to fixation alone (standard IHC practice). Use a no-primary control to identify signal introduced by the detection workflow (standard IHC practice).
How should I assess cytoplasmic or nucleolar SSRP1 staining?
Score the dominant chromogenic signal in nuclei: SSRP1 is reported in the nucleus and on chromatin, and HPA describes nuclear expression in most tissues (UniProt Q08945 subcellular location; HPA tissue IHC). Nucleolar staining can also be plausible because both UniProt and HPA report nucleolar localization (UniProt Q08945 subcellular location; HPA subcellular). A strong cytoplasmic pattern without corresponding nuclear staining deserves review against a no-primary control and adjacent morphology (UniProt Q08945 subcellular location; standard IHC practice). SSRP1 has no annotated transmembrane segment, so a crisp membrane rim is unexpected (UniProt Q08945 topology). Compare the suspect pattern with the same slide’s intact nuclei before assigning positive cells (standard IHC practice).
Can epitope masking or isoforms explain discordant SSRP1 staining?
The supplied UniProt record lists 0 isoforms, so an isoform-specific explanation is unsupported by this evidence (UniProt Q08945 isoforms). SSRP1 has annotated phosphorylation and acetylation sites, but the payload does not identify the A02606-1 epitope or show that these modifications affect antibody binding (UniProt Q08945 modified residues; caption A02606-1). If staining differs between blocks, first compare retrieval, fixation records, and intact nuclear morphology under matched staining conditions (standard IHC practice). Repeat weak sections with the documented EDTA pH 8.0 retrieval before attributing a difference to epitope biology (datasheet A02606-1; standard IHC practice). An independent antibody with a documented, distinct epitope can help investigate persistent discordance (standard IHC practice).
How can IF help investigate an ambiguous chromogenic SSRP1 pattern?
Use IF as a separate localization check: HPA reports mainly nucleoplasmic SSRP1, with additional nucleolar localization (HPA subcellular). Multiplex an SSRP1 channel with a marker for the expected cell type, then compare both channels against nuclear counterstaining at the single-cell level (standard IF practice). Choose a fluorophore whose emission is separated from the specimen’s autofluorescence, and include an unstained section to identify that background (standard IF practice). Because SSRP1 is nuclear and has no transmembrane segment, permeabilization should permit antibody access to the nucleus rather than target a membrane-facing epitope (UniProt Q08945 subcellular location and topology; standard IF practice). Optimize fixation and permeabilization for the IF specimen itself; the paraffin IHC caption does not establish IF conditions (caption A02606-1; standard IF practice).
What should I check when DAB background obscures SSRP1 nuclei?
First distinguish diffuse DAB deposition from the expected predominantly nuclear SSRP1 pattern by examining intact cells at higher magnification (HPA tissue IHC; standard IHC practice). Run a no-primary control and check peroxidase blocking, wash steps, and DAB development on the same staining run (standard IHC practice). The selected example used 10% goat serum block, 2 μg/ml primary overnight at 4°C, and a peroxidase-linked secondary before DAB development (caption A02606-1). If controls show background, adjust one variable at a time, including primary concentration or development time, while preserving a nuclear positive reference (standard IHC practice). Avoid interpreting deposits at folds or section edges as cellular signal (standard IHC practice).
How should I quantify SSRP1 across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment as the nucleus and set positivity thresholds using a consistent control and chromogen exposure across the batch (HPA tissue IHC; standard IHC practice). For each annotated cell population, report the percentage of positive nuclei and an H-score based on intensity categories 0–3 (standard IHC practice). Normalize positive counts to the number of evaluable nuclei in that population; if spatial abundance matters, report positive nuclei per mm² of viable tissue separately (standard IHC practice). Exclude necrotic, folded, and poorly preserved regions before scoring, and document the excluded area (standard IHC practice). Keep cell populations separate because HPA reports differing SSRP1 staining levels among cell types (HPA tissue IHC).
How do I distinguish true SSRP1 positivity from staining artefact?
A credible positive pattern is predominantly nuclear in intact cells; nucleolar signal is also compatible with reported SSRP1 localization (UniProt Q08945 subcellular location; HPA subcellular). Judge cell identity before calling a region positive: HPA reports medium staining in appendix lymphoid tissue and no detected staining in adipocytes (HPA tissue IHC). Treat isolated edge staining, necrotic-area deposits, or signal present in the no-primary control as possible artefacts requiring review (standard IHC practice). Check endogenous peroxidase blocking when DAB signal appears outside plausible cellular compartments (standard IHC practice). HPA rates the tissue-IHC antibody approved while noting low consistency with RNA expression, so interpret discordant results with that limitation in mind (HPA tissue IHC).
Boster reagents
Best SSRP1 / FACT complex subunit SSRP1 IHC Antibodies
A02606-1 has human paraffin-section IHC images from four cancer tissues (IHC image captions) and HeLa-cell IF data (IF image caption).
Real IHC data
Anti-SSRP1 Antibody ®
Cat # A02606-1
A02606-1 is shown in paraffin sections of human appendix adenocarcinoma, breast cancer, colon adenocarcinoma, and penis squamous cell carcinoma (IHC image captions). A02606-1 is also shown in HeLa cells by IF and lists ICC and IF applications (IF image caption; catalog applications).
Which to pick: Choose A02606-1 for paraffin-section IHC: its images use EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (IHC image captions). For IF/ICC, A02606-1 has a HeLa-cell IF image and lists both applications (IF image caption; catalog applications). For other species, A02606-1 lists human, monkey, mouse, and rat reactivity, while its supplied IHC images show human tissue only; no clone is specified (catalog reactivity and clone field; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q08945 (SSRP1_HUMAN, FACT complex subunit SSRP1).
- Human Protein Atlas. SSRP1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SSRP1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli and centrosome..
- Human Protein Atlas. SSRP1 antibody validation summary (2 antibodies).
- SSRP1 silencing inhibits the proliferation and malignancy of human glioma cells via the MAPK signaling pathway. Oncology reports 2017 — PMC5780019.
- Prognostic value of histone chaperone FACT subunits expression in breast cancer. Breast cancer (Dove Medical Press) 2017 — PMC5422336.
- SSRP1 influences colorectal cancer cell growth and apoptosis via the AKT pathway. International journal of medical sciences 2019 — PMC6909804.
- Expression of FACT in mammalian tissues suggests its role in maintaining of undifferentiated state of cells. Oncotarget 2011 — PMC3248156.
- PubMed PMID:1372440 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:9489704 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SSRP1 (FACT complex subunit SSRP1). Expected localisation Predominantly nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02606-1). Validated IHC antibody: A02606-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human appendix adenocarcinoma tissue; fixative not specified (datasheet A02606-1); 2. Fixation: Image fixative and duration unreported (datasheet A02606-1); 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 A02606-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02606-1); 8. Primary antibody: Rabbit anti-SSRP1, 2-5 μg/ml (datasheet A02606-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02606-1). Watch for: Fixation: Keep paraffin tissue fixation consistent across samples and controls. (selected-SKU IHC image A02606-1); Caveat: Low agreement between staining and RNA data (HPA tissue IHC); Isoform / epitope: No isoforms annotated; chain spans aa 2–709 (UniProt); Regulation: Recruited to actively transcribed loci (UniProt). Evidence: UniProt Q08945, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A02606-1, PMC5780019, PMC5422336. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SSRP1/immunohistochemistry-ihc-guide.