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SRRM1 / Serine/arginine repetitive matrix protein 1 · IHC design guide
Design Immunohistochemistry for SRRM1
Plan chromogenic SRRM1 IHC-P around its ubiquitous nuclear tissue pattern (HPA tissue IHC). This guide covers fixation consistency, antibody titration within 1:50–1:200 (datasheet A07296), and comparison with low-staining cardiomyocytes and fibroblasts (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SRRM1 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 in tissue (HPA tissue IHC); nuclear matrix and speckles (UniProt) | |
| Staining pattern | Ubiquitous nuclear staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A07296) | |
| 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 | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Cardiomyocytes and fibroblasts show low staining (HPA tissue IHC) | |
| Regulation | Broad tissue expression (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; epitope differences are unspecified (UniProt) |
Section 1
Recommended SRRM1 IHC & IF Protocols
The catalog antibody protocol is followed by published SRRM1 IHC methods for prostate and pancreatic cancer sections (PMC7000340; PMC13185238).
| Sample | Paraffin-embedded human esophageal cancer tissue; fixative not specified (datasheet A07296) |
| Fixation | Image fixative and duration unreported (datasheet A07296); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A07296); 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-SRRM1, 1:50-1:200 (datasheet A07296) |
| 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 | SRRM1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6.0 (datasheet A07296); the pancreatic study reports retrieval without specifying its conditions (PMC13185238).
Section 2
What Is the Expected SRRM1 Staining Pattern?
SRRM1 is a nuclear matrix and speckle protein with no transmembrane segment (UniProt Q8IYB3: subcellular location and topology). In paraffin section IHC, expect nuclear staining across many cell types, including glandular, respiratory epithelial and neuronal cells reported as high by HPA (HPA tissue IHC: ubiquitous nuclear expression; selected cells High). HPA rates the tissue staining Supported, with medium consistency between antibody staining and RNA expression (HPA tissue IHC: reliability).
| Nuclear staining in appendix or colon glandular cells, bronchial respiratory epithelium, or cortical neurons (HPA tissue IHC: High in each listed cell type). | This matches the reported broad nuclear pattern (HPA tissue IHC: ubiquitous nuclear expression). Compare nuclei within the same section before judging intensity across tissues; HPA's High calls describe observed staining, while its Supported rating includes a medium consistency caveat (HPA tissue IHC: reliability). |
| Strong staining confined to cytoplasm, membranes or extracellular material, with little nuclear signal (UniProt Q8IYB3: nuclear location; no transmembrane segment). | The compartment disagrees with the expected SRRM1 location (UniProt Q8IYB3: nucleus matrix and speckle). Treat it as a possible staining artefact and inspect morphology and controls before assigning it to SRRM1 (general IHC practice). |
| Prominent staining in a restricted cell population while the expected nuclei nearby remain unstained (HPA tissue IHC: ubiquitous nuclear expression). | Consider antibody cross-reactivity or endogenous detection activity, especially if the signal is outside nuclei (general IHC practice; UniProt Q8IYB3: nuclear location). Cardiomyocytes and fibroblasts are reported Low, not negative, so staining in either cell type alone does not establish a false positive (HPA tissue IHC: Low in cardiomyocytes and fibroblasts). |
| A broad, hazy chromogenic deposit obscures nuclear borders throughout the section (general IHC practice). | The background prevents a reliable compartment call (general IHC practice). Check the no-primary control, blocking and wash steps, and whether detection chemistry produces background; assess SRRM1 only after nuclei can be distinguished from surrounding tissue (general IHC practice). |
| No detectable nuclear signal in a section containing appendix or colon glandular cells reported High by HPA (HPA tissue IHC: High in these cells). | First check whether the IHC run and positive control worked, then review the antibody's validated application and detection steps (general IHC practice). One blank section does not prove SRRM1 absence; the reference pattern has Supported reliability with medium staining–RNA consistency (HPA tissue IHC: reliability). |
💡Expected SRRM1 appearanceCall a positive result when nuclei show discernible chromogenic staining in expected cells, with readily detectable signal in an HPA High cell population; isolated strong cytoplasmic or extracellular staining without matching nuclei is suspect (HPA tissue IHC: ubiquitous nuclear expression and selected cells High; UniProt Q8IYB3: nuclear location; general IHC practice).
| Compartment and topology (UniProt Q8IYB3: nucleus matrix and speckle; no transmembrane segment). | Score nuclear localisation first; a membrane pattern has no support in this record (UniProt Q8IYB3: subcellular location and topology). Chromogenic section IHC may show nuclei without resolving individual speckles (general IHC practice). |
| Tissue pattern and confidence (HPA tissue IHC: ubiquitous nuclear expression; reliability Supported). | HPA lists High staining in several glandular, respiratory epithelial and neuronal populations, and Low staining in cardiomyocytes and fibroblasts (HPA tissue IHC: listed cells). Interpret intensity differences with the stated medium staining–RNA consistency in mind (HPA tissue IHC: reliability description). |
| Antibody validation (HPA antibodies: HPA049941 IHC Supported; HPA058612 IHC unlisted). | Only HPA049941 has an IHC validation status in the supplied antibody list; HPA058612's ICC status does not establish its paraffin section IHC performance (HPA antibodies: application statuses). Check the IHC validation of the antibody actually used (general IHC practice). |
| Isoforms and antigen identity (UniProt Q8IYB3: isoforms 1 and 2; residues 1–904 in the listed chain). | Two isoforms are recorded, but this payload gives no antibody epitope or isoform-specific staining claim (UniProt Q8IYB3: isoforms; HPA antibodies: supplied fields). Do not infer which isoform a stained nucleus contains. |
| IF/ICC cross-check: where should puncta appear? (HPA subcellular: nuclear speckles, enhanced). | In IF/ICC, HPA reports enhanced nuclear-speckle localisation; its image list includes HEK293, HeLa, U2OS and NIH 3T3 (HPA subcellular: location and image cell lines). This supports a localisation comparison, not an IHC protocol choice (HPA subcellular: ICC-IF evidence). |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported High population is blank on the IHC slide (HPA tissue IHC: listed High populations). | Possible run failure, insufficient detection or an antibody without established IHC performance (general IHC practice; HPA antibodies: application statuses). | Review the positive control, antibody application validation, detection reagents and counterstain; optimise the IHC workflow using the antibody's own instructions (general IHC practice). Do not assign a target-specific fixation cause from this record. |
| Signal is mainly cytoplasmic or membranous (UniProt Q8IYB3: nuclear location; no transmembrane segment). | Nonspecific binding or background may dominate the intended nuclear signal (general IHC practice). The observed compartment conflicts with the supplied SRRM1 localisation (UniProt Q8IYB3: subcellular location). | Inspect a no-primary control and tissue morphology, then review blocking, washes and detection settings (general IHC practice). Require convincing nuclear staining before scoring the section as SRRM1 positive (UniProt Q8IYB3: nuclear location). |
| Brown deposit follows blood-rich or otherwise reactive areas more than nuclei (general IHC practice). | Endogenous activity can contribute to background in enzyme-based chromogenic detection (general IHC practice). | Use a no-primary control; if detection is HRP based, review the endogenous peroxidase blocking step and confirm that the remaining signal is nuclear (general IHC practice; UniProt Q8IYB3: nuclear location). |
| Weak nuclear staining is seen in cardiomyocytes or fibroblasts (HPA tissue IHC: Low in these cells). | Low reference staining can yield a faint result; the supplied HPA record does not label either population negative (HPA tissue IHC: Low; negative list empty). | Interpret intensity alongside a reported High population in the same run and keep the Low category distinct from absence (HPA tissue IHC: High and Low cell lists; general IHC practice). |
| All structures have diffuse colour, making nuclear localisation unreadable (general IHC practice). | Broad background can obscure the compartment pattern; the appearance alone does not identify a target-specific cause (general IHC practice). | Compare with the no-primary control, review antibody concentration against its instructions, and check blocking and wash steps before scoring (general IHC practice). |
| IF/ICC shows nuclear speckles, but chromogenic IHC looks broadly nuclear (HPA subcellular: nuclear speckles; HPA tissue IHC: nuclear expression). | The two readouts can differ in visible spatial detail; the supplied records describe speckles for ICC-IF and nuclear expression for tissue IHC (HPA subcellular; HPA tissue IHC). | Judge paraffin section IHC against a nuclear pattern and appropriate controls; use the ICC-IF speckle result as a localisation cross-check, without requiring resolved puncta on the IHC slide (general IHC practice; HPA subcellular). |
Experiment evidence
IHC & IF images showing SRRM1 staining
A07296 · IHCImmunohistochemistry of paraffin-embedded human esophageal cancer using SRRM1 Rabbit pAb at dilution of 1:450 .Perform high pressure antigen
A07296 · IHCImmunohistochemistry of paraffin-embedded mouse testis using SRRM1 Rabbit pAb at dilution of 1:450 .Perform high pressure antigen retrieval
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000133226-SRRM1/tissue/cerebral+cortex#imid_13806842
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000133226-SRRM1/tissue/colon#imid_13788121
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000133226-SRRM1/tissue/liver#imid_13788114
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000133226-SRRM1/tissue/kidney#imid_13788107
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA049941; green = SRRM1, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA049941; green = SRRM1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SRRM1 IHC & IF
🧪Run appendix first and score its glandular cells for SRRM1 staining (HPA: High in appendix glandular cells). HPA detects SRRM1 in all 45 scored tissues, so there is no supported negative tissue or validated negative cell population within the appendix slide; use no-primary and isotype controls to define background, and expect specific signal to be nuclear rather than cytoplasmic (HPA: no negative tissue rows; HPA subcellular: nuclear speckles).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: None in HPA: SRRM1 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 SRRM1 in HEK293, HeLa, U2OS, NIH 3T3, with annotated localisation: Nuclear speckles (enhanced) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only control and a concentration-matched isotype control of the same host species and immunoglobulin class as the primary antibody; use SRRM1 knockout material or a validated peptide-block experiment as a biological specificity control. For chromogenic appendix IHC, quench endogenous peroxidase and check inflammatory cells for residual background; block endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the fixative in the selected A07296 paraffin-section caption is unreported (A07296 tissue-IHC caption). That caption uses high-pressure retrieval with 10 mM citrate, pH 6.0, at 1:450 in human esophageal cancer; whether retrieval is required for appendix or whether frozen sections or IF/ICC are easier is unreported (A07296 tissue-IHC caption). In appendix, distinguish glandular nuclear signal from luminal mucus and inflammatory-cell peroxidase background (HPA: High in appendix glandular cells; standard IHC practice).
HPA tissue IHC evidence for SRRM1
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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: SRRM1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced SRRM1 IHC Tips
Troubleshoot SRRM1 staining in paraffin sections by checking retrieval, nuclear localisation and controls before comparing staining across samples.
What retrieval should I try first when SRRM1 staining is weak?
Start with heat-mediated retrieval in 10 mM citrate buffer, pH 6.0, for paraffin sections (datasheet A07296; A07296 tissue-IHC caption). The selected image used high-pressure retrieval with that buffer and an antibody dilution of 1:450, providing a matched starting condition for this SKU (A07296 tissue-IHC caption). Keep heating, cooling and section thickness consistent while adjusting retrieval duration in small steps, then compare nuclear signal with tissue damage on adjacent sections (standard IHC practice). If staining remains weak, assess an alternative retrieval buffer only as a controlled fallback, alongside the citrate condition and a no-primary control (standard IHC practice).
Could fixation explain inconsistent SRRM1 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative, and the supplied evidence does not establish how fixation changes SRRM1 staining (A07296 tissue-IHC caption). Record each block's documented fixation and processing history before comparing signal, since those variables can affect antigen accessibility in IHC (standard IHC practice). Apply the same 10 mM citrate, pH 6.0 retrieval and detection conditions to comparison sections so processing differences are easier to assess (datasheet A07296; standard IHC practice). If intensity differs, check section integrity and nuclear counterstaining before attributing the difference to SRRM1 abundance (standard IHC practice).
Where should credible SRRM1 chromogenic staining appear?
Expect predominantly nuclear staining: SRRM1 is assigned to the nuclear matrix and speckles, while tissue IHC reports ubiquitous nuclear expression (UniProt Q8IYB3; HPA tissue IHC). HPA subcellular imaging further places SRRM1 in nuclear speckles, although punctate detail may be harder to resolve with chromogenic tissue IHC (HPA subcellular; standard IHC practice). Compare brown signal with the nuclear counterstain at high magnification, scoring cells only when the signal can be assigned to nuclei (standard IHC practice). Predominantly membranous or diffuse extracellular colour conflicts with the documented localisation and warrants inspection of background and detection controls (UniProt Q8IYB3; standard IHC practice).
How can epitope choice complicate interpretation of SRRM1 staining?
SRRM1 has 2 annotated isoforms, a PWI domain at residues 27–126, and numerous modified residues; the supplied material does not map this antibody's epitope (UniProt Q8IYB3; A07296 tissue-IHC caption). Therefore, the observed nuclear stain cannot establish which isoform contributes to the signal or whether a modification changes antibody binding (UniProt Q8IYB3; standard IHC interpretation). Obtain an epitope map or validation data before making either claim, and compare samples under matched retrieval conditions (standard IHC practice). If staining differs unexpectedly, use an independently validated antibody to a distinct epitope as an orthogonal check, while retaining a no-primary control for detection background (standard IHC practice).
How should I design an IF follow-up to the SRRM1 tissue IHC result?
Treat IF as a separately optimised assay: the supplied A07296 image documents chromogenic staining of paraffin sections, and HPA reports nuclear-speckle localisation from subcellular imaging (A07296 tissue-IHC caption; HPA subcellular). Multiplex SRRM1 with a marker for the cell type being evaluated and a nuclear stain, selecting antibodies and detection channels that can be distinguished reliably (standard IF practice). Choose a fluorophore whose emission can be separated from the specimen's measured autofluorescence, and include single-label and no-primary controls (standard IF practice). SRRM1 has no transmembrane segment and is nuclear, so optimise permeabilisation for access to an intracellular nuclear epitope without assuming an antibody-specific requirement (UniProt Q8IYB3; standard IF practice).
What should I check when SRRM1 IHC produces diffuse brown background?
First compare a no-primary section with the stained section to identify colour generated by detection chemistry rather than the primary antibody (standard IHC practice). For a peroxidase and DAB workflow, verify the peroxidase block, blocking conditions, washes and development time; these are general workflow controls, not demonstrated SRRM1-specific remedies (standard IHC practice). Titrate the catalog antibody around the caption's 1:450 condition while keeping retrieval at 10 mM citrate, pH 6.0, and assess nuclear contrast rather than overall darkness (A07296 tissue-IHC caption; standard IHC practice). Diffuse extracellular or predominantly membranous colour is inconsistent with the documented nuclear localisation and should prompt review of tissue quality and controls (UniProt Q8IYB3; HPA tissue IHC).
How should I quantify SRRM1 across differently cellular tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis regions before scoring, then count nuclei with interpretable SRRM1 signal against the total eligible nuclei in each region (standard IHC practice). Report % positive nuclei with a fixed positivity threshold, or use a nuclear H-score when intensity differences are reproducible under matched staining and imaging conditions (standard IHC practice). Normalise positive-cell counts to eligible nuclei; if reporting positive-cell density, divide by analysed area in mm² and also report cellularity (standard IHC practice). Keep cell populations separate where possible, since HPA reports ubiquitous nuclear expression but differing staining levels among listed tissue cell types (HPA tissue IHC).
How do I distinguish convincing SRRM1 positivity from staining artefacts?
Look for nuclear staining within intact cells, consistent with SRRM1's nuclear matrix and speckle localisation and HPA's ubiquitous nuclear tissue profile (UniProt Q8IYB3; HPA tissue IHC). Check that stained nuclei belong to the cell population being interpreted; HPA lists high staining in several specified cell types and low staining in cardiomyocytes and fibroblasts (HPA tissue IHC). Discount signal concentrated at section edges or necrotic areas when it does not follow intact nuclei, and compare with adjacent well-preserved regions (standard IHC practice). Brown colour retained in a no-primary control can indicate endogenous enzyme or detection background and should not be scored as SRRM1 positivity (standard IHC practice).
Boster reagents
Best SRRM1 / Serine/arginine repetitive matrix protein 1 IHC Antibodies
A07296 has IHC images from paraffin-embedded human esophageal cancer and mouse testis; no IF image is supplied (A07296 IHC image captions; catalog: no IF images).
Real IHC data
Anti-SRRM1 Antibody
Cat # A07296
A07296 is listed for human and mouse IHC (catalog: applications and reactivity). Its IHC images show staining of paraffin-embedded human esophageal cancer and mouse testis (A07296 IHC image captions).
Which to pick: For tissue IHC and human–mouse work, choose the rabbit polyclonal A07296: IHC and both species are listed, and its images show paraffin sections retrieved under high pressure in 10 mM citrate buffer, pH 6.0 (catalog: host, applications and reactivity; A07296 IHC image captions). The captions do not report a fixative (A07296 IHC image captions). For IF/ICC, A07296 has no listed IF application or IF image, so the payload does not establish an IF/ICC choice (catalog: applications and IF images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IYB3 (SRRM1_HUMAN, Serine/arginine repetitive matrix protein 1).
- Human Protein Atlas. SRRM1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. SRRM1 subcellular location (ICC-IF): Localized to the nuclear speckles..
- Human Protein Atlas. SRRM1 antibody validation summary (2 antibodies).
- Dysregulation of the splicing machinery is directly associated to aggressiveness of prostate cancer. EBioMedicine 2020 — PMC7000340.
- Combining ferroptosis inducers with gemcitabine to enhance treatment efficacy in pancreatic cancer. Cancer & metabolism 2026 — PMC13185238.
- Clinical value of circulating splicing factors in prostate cancer: SRRM1 as a novel predictive biomarker and therapeutic target. Molecular therapy. Oncology 2024 — PMC11697196.
- PubMed PMID:9531537 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SRRM1 (Serine/arginine repetitive matrix protein 1). Expected localisation Nuclear in tissue (HPA tissue IHC); nuclear matrix and speckles (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A07296). Validated IHC antibody: A07296 at 1:50-1:200 (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: Paraffin-embedded human esophageal cancer tissue; fixative not specified (datasheet A07296); 2. Fixation: Image fixative and duration unreported (datasheet A07296); 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: Citrate pH 6.0 (datasheet A07296); 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-SRRM1, 1:50-1:200 (datasheet A07296); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Cardiomyocytes and fibroblasts show low staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences are unspecified (UniProt); Regulation: Broad tissue expression (HPA tissue IHC). Evidence: UniProt Q8IYB3, HPA tissue IHC · Supported, HPA ICC-IF · nuclear speckles., Datasheet A07296, PMC7000340, PMC13185238. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SRRM1/immunohistochemistry-ihc-guide.