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SRRT / Serrate RNA effector molecule homolog · IHC design guide
Design Immunohistochemistry for SRRT
Plan SRRT staining in paraffin sections with the catalog antibody at 2–5 μg/ml (datasheet A05444-1). This guide helps assess the expected nuclear pattern across tissues (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SRRT 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 | Ubiquitous nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in glandular, neuronal and hematopoietic cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05444-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Soft tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Ubiquitously expressed (UniProt) | |
| Isoform / epitope | 5 isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended SRRT IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet: A05444-1). The published IHC protocols below describe SRRT staining in prostate and head and neck cancer tissues (PMC10216025; PMC7905686; PMC13195263).
| Sample | Paraffin-embedded human lung adenocarcinoma tissue; fixative not specified (datasheet A05444-1) |
| Fixation | Image fixative and duration unreported (datasheet A05444-1); 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 A05444-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05444-1) |
| Primary antibody | Rabbit anti-SRRT, 2-5 μg/ml (datasheet A05444-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05444-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05444-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SRRT-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet: A05444-1). Citrate retrieval is reported with other SRRT antibodies (PMC7905686; PMC13195263).
Section 2
What Is the Expected SRRT Staining Pattern?
SRRT is predominantly nuclear, with nucleoplasmic localisation and possible cytoplasmic shuttling (UniProt Q9BXP5; HPA: enhanced nucleoplasm in ICC-IF). In paraffin-section IHC, expect nuclear staining across many cell types, including HPA-reported high-staining glandular, hematopoietic and neuronal cells (HPA: ubiquitous nuclear expression; listed tissue IHC). The tissue profile is Supported, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability). SRRT has no transmembrane segment (UniProt Q9BXP5 topology).
| Distinct nuclear staining in glandular cells of colon, breast or adrenal gland, or in bone-marrow hematopoietic cells. | This fits the reported high-staining cell populations and ubiquitous nuclear profile (HPA: tissue IHC). Judge the relevant cells against neighbouring tissue and the counterstain; staining need not be identical in every nucleus (general IHC practice). |
| Cytoplasmic signal dominates while nuclei are consistently pale in an otherwise positive tissue. | Treat this as a discordant compartment pattern requiring controls, rather than an expected IHC result: SRRT is predominantly nuclear, although it can shuttle to the cytoplasm (UniProt Q9BXP5). HPA reports ubiquitous nuclear tissue staining (HPA: tissue IHC). |
| Strong staining appears chiefly in peripheral nerve within soft tissue, while expected positive cells are unstained. | Investigate cross-reactivity or endogenous detection activity (general IHC practice). Peripheral nerve was reported as not detected in the sampled soft-tissue IHC, but that observation is not a universal negative-control guarantee (HPA: soft-tissue IHC). |
| Brown chromogen spreads across nuclei, cytoplasm and extracellular areas without clear cell boundaries. | This is diffuse background rather than the reported nuclear pattern (HPA: tissue IHC). Check reagent-only controls, blocking, washes and detection conditions before assigning SRRT positivity (general IHC practice). |
| No nuclear signal is seen in a high-staining reference cell population. | The run may have failed, or the chosen antibody conditions may be unsuitable (general IHC practice). Confirm the reference population and section quality before interpreting other cells as negative (HPA: high glandular, hematopoietic and neuronal IHC staining). |
💡Expected SRRT appearanceCall a result positive when staining is predominantly nuclear in an HPA-reported high-staining cell population; diffuse extracellular colour or cytoplasm-dominant staining without matching nuclei should trigger a control review (HPA: tissue IHC; UniProt Q9BXP5 localisation; general IHC practice).
| Compartment and cell context | Score nuclear staining within identifiable cells; SRRT is predominantly nuclear (UniProt Q9BXP5), and HPA describes ubiquitous nuclear tissue expression with high staining in several listed cell populations (HPA: tissue IHC). |
| Antibody validation | HPA042858 has Supported IHC status; HPA058379 has no listed IHC status (HPA: antibody validation). ICC-IF validation for either antibody does not, by itself, establish its paraffin-section IHC performance (HPA: antibody validation; general assay distinction). |
| Negative tissue comparison | Peripheral nerve in sampled soft tissue was not detected, whereas listed glandular, hematopoietic and neuronal populations stained High (HPA: tissue IHC). Compare the specified cells, not whole-section colour; SRRT is ubiquitously expressed (UniProt Q9BXP5). |
| Isoforms and epitope coverage | Five isoforms are listed (UniProt Q9BXP5). Their existence alone does not show which forms an antibody recognises; consult the antibody's stated immunogen or epitope before attributing a staining difference to isoform expression (general IHC practice). |
| IF/ICC comparison: should the IHC pattern be identical? | Use nucleoplasmic IF as a localisation cross-check, not an IHC protocol: HPA reports enhanced nucleoplasmic ICC-IF localisation (HPA: subcellular ICC-IF). Compare compartment calls while accounting for distinct specimen preparation and detection (general IHC/IF practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells show no nuclear chromogen. | A failed staining run or unsuitable antibody conditions are possible (general IHC practice); listed high-staining cells provide a comparison (HPA: tissue IHC). | Check a concurrently processed high-staining reference, section integrity, antibody application and detection controls; then review the antibody's IHC instructions (general IHC practice). |
| Signal is mainly cytoplasmic across the section. | The dominant pattern conflicts with reported nuclear tissue staining, although cytoplasmic shuttling is described (HPA: tissue IHC; UniProt Q9BXP5). | Compare with counterstained nuclei and a positive reference; check background and reagent-only controls before accepting a cytoplasmic interpretation (general IHC practice). |
| Peripheral nerve stains strongly in sampled soft tissue. | This differs from HPA's not-detected observation; cross-reactivity or endogenous detection activity is possible (HPA: soft-tissue IHC; general IHC practice). | Verify the cell identity and run an appropriate reagent-only control; interpret the discrepancy alongside an HPA-reported positive population (general IHC practice; HPA: tissue IHC). |
| Diffuse chromogen obscures the nuclear pattern. | Excess background can arise from blocking, washing or detection conditions (general IHC practice). | Review reagent-only controls, blocking and washes, then reassess whether nuclear signal remains above local background (general IHC practice; HPA: nuclear tissue profile). |
| Only some expected cells stain, or intensity varies between regions. | HPA lists specific high-staining cell populations, while its overall tissue reliability is Supported with medium RNA agreement (HPA: tissue IHC). | Score defined cell populations and nuclear localisation separately from overall tissue colour; record regional variation without treating every pale cell as a technical failure (general IHC practice). |
| ICC-IF appears nucleoplasmic, but the paraffin-section IHC result is unclear. | The assays use different preparation and detection methods (general IHC/IF practice); HPA reports enhanced nucleoplasmic ICC-IF localisation (HPA: subcellular ICC-IF). | Use the IF result to check the expected compartment, then resolve IHC with its own positive reference and detection controls (general IHC practice). |
Experiment evidence
IHC & IF images showing SRRT staining
A05444-1 · IHCIHC analysis of ARS2/SRRT using anti-ARS2/SRRT antibody (A05444-1). ARS2/SRRT was detected in a paraffin-embedded section of human lung aden
A05444-1 · IHCIHC analysis of ARS2/SRRT using anti-ARS2/SRRT antibody (A05444-1). ARS2/SRRT was detected in a paraffin-embedded section of human appendix
A05444-1 · IFIF analysis of ARS2/SRRT using anti-ARS2/SRRT antibody (A05444-1) and anti-Tubulin Alpha antibody (M03989-3). ARS2/SRRT was detected in immu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000087087-SRRT/tissue/cerebral+cortex#imid_13159671
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000087087-SRRT/tissue/colon#imid_13159605
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000087087-SRRT/tissue/liver#imid_13159590
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000087087-SRRT/tissue/kidney#imid_13159599
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA042858; green = SRRT, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA042858; green = SRRT, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SRRT IHC & IF
🧪Run adrenal gland first and score its glandular cells for SRRT staining (HPA: High in adrenal gland glandular cells). Use peripheral nerve within soft tissue as the negative comparison (HPA: Not detected in soft tissue peripheral nerve); on the adrenal slide, anucleate erythrocytes, if present, should show no nuclear signal (standard IHC practice), while HPA identifies no SRRT-negative nucleated cell type there (HPA: adrenal gland row).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Soft tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SRRT in HEK293, MCF-7, U2OS, U2OS, siRNA 1 (10x), U2OS, siRNA 2 (10x), U2OS, scrambled (10x), NIH 3T3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a concentration-matched rabbit IgG isotype control (selected A05444-1 caption: rabbit primary); use an SRRT knockout specimen as a biological negative where available (standard IHC practice). Block endogenous peroxidase and inspect blood-rich areas for residual DAB background (selected A05444-1 caption: peroxidase detection with DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state a fixative (selected A05444-1 caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 was used, but whether SRRT detection depends on that retrieval condition is unreported (selected A05444-1 caption). Paraffin-section IHC has a documented workflow, while the supplied evidence does not establish that frozen sections or IF are easier; assess blood-associated peroxidase background in adrenal sections (selected A05444-1 caption; HPA: High in adrenal gland glandular cells; standard IHC practice).
HPA tissue IHC evidence for SRRT
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Basal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | Peripheral nerve | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SRRT IHC Tips
Use nuclear staining as the primary readout for SRRT in paraffin section IHC, with controls that distinguish signal from tissue artefact.
Which retrieval condition should I start with for SRRT in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A05444-1). The selected antibody was used on a paraffin section after this retrieval, followed by 10% goat serum blocking and 2 µg/mL primary antibody overnight at 4°C (datasheet A05444-1). If nuclear staining is weak, first check heating consistency and section adherence; then compare a second retrieval condition on matched sections as an optimisation experiment (standard IHC practice). Judge improvement by nuclear signal in intact cells and by background on a no primary control, rather than total DAB intensity alone (HPA tissue IHC; standard IHC practice).
How should I handle uncertain fixation effects on SRRT staining?
The selected tissue caption identifies a paraffin section but does not state its fixative, so target specific fixation sensitivity is unknown (datasheet A05444-1). Record the fixative, fixation duration, section age and retrieval conditions for each specimen, and compare matched sections when optimising staining (standard IHC practice). Keep the documented EDTA pH 8.0 retrieval and 2 µg/mL primary concentration constant during the fixation comparison (datasheet A05444-1). Score intact nuclear staining alongside tissue morphology and no primary background, since apparent loss of signal in damaged areas cannot establish a specific fixation effect on SRRT (HPA tissue IHC; standard IHC practice).
What staining pattern should count as plausible SRRT localisation?
Prioritise staining in nuclei, especially the nucleoplasm, because tissue IHC describes ubiquitous nuclear expression and cell imaging places SRRT in the nucleoplasm (HPA tissue IHC; HPA subcellular). SRRT is predominantly nuclear but can shuttle to the cytoplasm, so a reproducible cytoplasmic component warrants review rather than automatic exclusion (UniProt Q9BXP5 localisation). Compare cells within the same section and check whether the nuclear compartment remains clearly distinguishable from DAB outside it (standard IHC practice). Strong staining confined to extracellular material or cell borders is inconsistent with the reported localisation and should prompt review of morphology and control sections (UniProt Q9BXP5 localisation; standard IHC practice).
Can this antibody distinguish SRRT isoforms or modification states in tissue?
SRRT has 5 annotated isoforms and multiple modified residues, including phosphorylation sites, but the supplied tissue caption does not map the antibody epitope (UniProt Q9BXP5 isoforms and modified residues; datasheet A05444-1). Treat chromogenic signal as SRRT associated staining without assigning an isoform or phosphorylation state (standard IHC interpretation). If that distinction matters, obtain epitope documentation and compare it with isoform sequences before interpreting staining differences (standard antibody validation practice). Keep retrieval and detection conditions matched across specimens, since a change in DAB intensity alone cannot identify an isoform or modification (standard IHC practice).
How would I check SRRT localisation by multiplex IF?
Use the separate IF/ICC workflow to compare SRRT with a validated marker for the expected cell type and a nuclear counterstain; nucleoplasmic enrichment is the anticipated pattern (HPA subcellular; standard IF practice). In glandular tissue, a glandular cell marker can help assign the signal to the relevant cells (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before interpreting overlap, with a longer wavelength channel if shorter wavelength background is high (standard IF practice). Because SRRT is nucleoplasmic and has no transmembrane segment, optimise permeabilisation for access to intracellular and nuclear epitopes while checking that morphology remains intact (UniProt Q9BXP5 localisation and topology; standard IF practice).
How can I separate SRRT signal from chromogenic background?
Examine a no primary control beside the stained section to identify DAB deposition from the detection workflow, and include a peroxidase blocking step appropriate to the tissue (standard chromogenic IHC practice). The selected caption used a peroxidase conjugated secondary antibody and DAB, so diffuse brown signal should be assessed against that detection chemistry (datasheet A05444-1). Inspect section edges, folds and damaged regions separately from intact nuclei because uneven reagent exposure can exaggerate staining there (standard IHC practice). If background persists, reassess blocking, washing and primary concentration against the documented 10% goat serum block and 2 µg/mL primary condition (datasheet A05444-1; standard IHC practice).
How should I quantify SRRT staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score SRRT within morphologically intact nuclei, using the same cell population and sampling rule across sections (HPA tissue IHC; standard IHC practice). Report the percentage of positive nuclei and, when intensity is reproducible, an H-score calculated as the sum of each intensity category’s percentage multiplied by its category value (standard IHC scoring practice). Normalise positive counts to all evaluable nuclei in the region, or report positive nuclear density per mm² when cell density itself matters (standard IHC quantification practice). Exclude folds, necrosis and section edges by a prespecified rule, and retain the same DAB development and imaging settings for comparisons (standard IHC practice).
Which findings would make an apparent SRRT positive result doubtful?
A credible result shows staining in intact nuclei, consistent with SRRT’s predominantly nuclear localisation and the ubiquitous nuclear tissue profile (UniProt Q9BXP5 localisation; HPA tissue IHC). Staining restricted to extracellular spaces, necrotic areas or section edges is suspect, especially when intact nuclei nearby are unstained (UniProt Q9BXP5 localisation; standard IHC practice). Check cell identity before calling a compartment unexpected: HPA reports high staining in several glandular cell populations, hematopoietic cells and neuronal cells, while peripheral nerve in soft tissue was not detected (HPA tissue IHC). Compare the no primary control and peroxidase block to assess endogenous enzyme signal before attributing brown deposits to SRRT (standard chromogenic IHC practice).
Boster reagents
Best SRRT / Serrate RNA effector molecule homolog IHC Antibodies
The catalog antibody has paraffin-section IHC images from human tumors and an IF/ICC image from A549 cells (catalog image captions); its listed reactivity covers human, mouse and rat (catalog reactivity).
Real IHC data
Anti-ARS2/SRRT Antibody ®
Cat # A05444-1
A05444-1 has IHC images from paraffin-embedded human lung adenocarcinoma, appendix adenocarcinoma, breast cancer and endometrioid adenocarcinoma sections (A05444-1 IHC captions). The same SKU has an IF/ICC image from A549 cells (A05444-1 IF caption) and lists human, mouse and rat reactivity (catalog reactivity).
Which to pick: Choose A05444-1 for paraffin-section IHC: its own human tissue captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml (A05444-1 IHC captions); the fixative is unreported (A05444-1 IHC captions). For IF/ICC, A05444-1 has an A549 image using 5 μg/ml primary antibody (A05444-1 IF caption). For mouse or rat samples, A05444-1 lists reactivity with both species, although the supplied IHC and IF images show human samples (catalog reactivity; A05444-1 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BXP5 (SRRT_HUMAN, Serrate RNA effector molecule homolog).
- Human Protein Atlas. SRRT tissue IHC expression (reliability: Supported).
- Human Protein Atlas. SRRT subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. SRRT antibody validation summary (2 antibodies).
- Serrate RNA Effector Molecule (SRRT) Is Associated with Prostate Cancer Progression and Is a Predictor of Poor Prognosis in Lethal Prostate Cancer. Cancers 2023 — PMC10216025.
- PSMC2, ORC5 and KRTDAP are specific biomarkers for HPV-negative head and neck squamous cell carcinoma. Oncology letters 2021 — PMC7905686.
- SRRT promotes prostate cancer progression and serves as a prognostic biomarker through STAT3 pathway activation. Oncology reports 2026 — PMC13195263.
- PubMed PMID:11239002 — UniProt-cited evidence.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SRRT (Serrate RNA effector molecule homolog). Expected localisation Ubiquitous nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05444-1). Validated IHC antibody: A05444-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Soft tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung adenocarcinoma tissue; fixative not specified (datasheet A05444-1); 2. Fixation: Image fixative and duration unreported (datasheet A05444-1); 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 A05444-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05444-1); 8. Primary antibody: Rabbit anti-SRRT, 2-5 μg/ml (datasheet A05444-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05444-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 5 isoforms; check antibody epitope coverage (UniProt); Regulation: Ubiquitously expressed (UniProt). Evidence: UniProt Q9BXP5, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A05444-1, PMC10216025, PMC7905686. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SRRT/immunohistochemistry-ihc-guide.