This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
RSRC1 / Serine/Arginine-related protein 53 · IHC design guide
Design Immunohistochemistry for RSRC1
Use nuclear and cytoplasmic staining as the RSRC1 tissue IHC benchmark (HPA tissue IHC). This guide highlights glandular cells with high staining (HPA tissue IHC) and the catalog antibody’s 5 μg/mL IHC starting concentration (datasheet: A12655).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RSRC1 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 and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Nuclear and cytoplasmic staining; high in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Duodenum+4 more · see all | |
| Negative control | Adipose 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 and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Widely expressed; regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; verify antibody epitope coverage (UniProt) |
Section 1
Recommended RSRC1 IHC & IF Protocols
This section pairs the catalog antibody’s IHC-P protocol with one published RSRC1 staining method (PMC6625388).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A12655); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-RSRC1, 5 μg/mL (datasheet A12655) |
| 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 | RSRC1-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: Nuclear and cytoplasmic expression in all tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval rule; UniProt: nucleus).
Section 2
What Is the Expected RSRC1 Staining Pattern?
RSRC1 is nuclear, including nuclear speckles, and can also be cytoplasmic (UniProt Q96IZ7; HPA tissue IHC; HPA ICC-IF). Expect staining across many cell types, with strong examples in glandular cells of duodenum, fallopian tube and gallbladder, and in testicular spermatogonia (HPA: High). HPA rates tissue IHC as Supported, with medium consistency between staining and RNA data (HPA tissue IHC). RSRC1 has no transmembrane segment (UniProt Q96IZ7 topology).
| Nuclear and cytoplasmic chromogen in glandular cells of duodenum, fallopian tube or gallbladder. | This fits the reported distribution and strong cell examples (HPA tissue IHC: nuclear and cytoplasmic; HPA: High in these glandular cells). Judge localisation within identifiable cells, because broad tissue expression does not mean every cell must stain equally (HPA: low tissue specificity; general IHC interpretation). |
| A crisp nuclear-speckle pattern, with or without detectable cytoplasmic staining. | Nuclear speckles are the approved ICC-IF location (HPA subcellular ICC-IF). UniProt also places RSRC1 in the nucleus and cytoplasm and describes shuttling (UniProt Q96IZ7). The speckle finding helps assess localisation, but an IHC section need not resolve discrete speckles as clearly as ICC-IF (general microscopy practice). |
| Predominantly cell-surface or extracellular staining, without a credible nuclear component. | That compartment is discordant with the reported nuclear and cytoplasmic pattern (HPA tissue IHC; UniProt Q96IZ7) and the absence of a transmembrane segment (UniProt Q96IZ7 topology). Treat it as a possible artefact; inspect morphology and detection controls before assigning RSRC1 localisation (general IHC practice). |
| Strong signal confined to adipocytes while expected positive cells remain blank. | Adipocytes are listed as Not detected, whereas several glandular populations and spermatogonia are High (HPA tissue IHC). This reversal raises cross-reactivity or endogenous detection activity as possibilities, not a diagnosis; compare an appropriate negative control and a known-positive section (general IHC practice). |
| Diffuse colour obscures cell boundaries, or no signal appears in a known-positive section. | Diffuse background cannot establish the reported intracellular localisation (HPA tissue IHC; general IHC interpretation). A blank known-positive section leaves the run uninterpretable: check the staining controls and workflow before inferring absent RSRC1, since HPA documents High staining in specified cells (HPA tissue IHC; general IHC practice). |
💡Expected RSRC1 appearanceCall a section positive when identifiable glandular cells in an HPA High tissue show strong nuclear and/or cytoplasmic staining (HPA tissue IHC: High; nuclear and cytoplasmic), while a membrane-only deposit or strong adipocyte-only signal warrants scrutiny rather than an RSRC1-positive call (UniProt Q96IZ7 topology; HPA: adipocytes Not detected; general IHC interpretation).
| Tissue and cell selection | Use the named High populations to assess an IHC run; hepatocytes and breast glandular cells are Low, and adipocytes are Not detected (HPA tissue IHC). A weak result in a Low population is therefore less informative than a failed High-cell control (HPA tissue IHC; general IHC interpretation). |
| Validation scope | HPA rates tissue IHC Supported and reports medium consistency with RNA; HPA044792 is Supported for IHC and Approved for ICC (HPA tissue IHC; HPA antibodies). These ratings support pattern assessment, but do not establish fixation sensitivity, retrieval conditions or performance for another antibody (HPA validation scope). |
| Isoforms and epitope coverage | Two isoforms are listed (UniProt Q96IZ7). Their listing alone does not show which isoforms an antibody detects; consult the chosen antibody's documented epitope before attributing a staining difference to isoforms (UniProt Q96IZ7; general antibody interpretation). |
| IF/ICC Q: What localisation should an image show? | A: Nuclear speckles are the approved main location (HPA subcellular ICC-IF). This answers localisation only; the separate IF/ICC guide should supply its own staining workflow (HPA subcellular ICC-IF; application scope). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive glandular cells show no chromogen. | A failed staining run is possible; those cells are reported High (HPA tissue IHC). | Confirm tissue identity and cell preservation, then check the positive control, primary-antibody step, retrieval and detection reagents according to the assay workflow (general IHC practice). Do not assign an RSRC1-specific fixation or retrieval cause from these sources (HPA/UniProt evidence scope). |
| Signal is faint in liver hepatocytes or breast glandular cells. | Both populations are listed Low (HPA tissue IHC). | Compare a named High-cell section processed in the same run before changing assay conditions (HPA tissue IHC; general IHC practice). Score the relevant cells rather than treating a weak Low-cell result alone as a failed antibody (general IHC interpretation). |
| Brown deposit appears mainly at cell borders or outside cells. | This distribution conflicts with nuclear and cytoplasmic RSRC1 and its lack of a transmembrane segment (HPA tissue IHC; UniProt Q96IZ7 topology). | Inspect tissue morphology and a primary-omission control, then review washing and detection steps (general IHC practice). Require intracellular signal in the expected cells before calling it specific (HPA tissue IHC; general IHC interpretation). |
| Adipocytes stain strongly. | HPA lists adipocytes as Not detected; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Compare a primary-omission control and a named High-cell positive section; review detection blocking as appropriate for the chemistry used (general IHC practice). Avoid using adipocyte colour alone to score RSRC1 (HPA tissue IHC). |
| Background makes nuclear and cytoplasmic staining hard to judge. | Nonspecific binding or detection background may obscure cell-level localisation (general IHC practice). | Review blocking, primary-antibody dilution, washing and counterstain against the assay instructions; compare controls before adjusting one condition at a time (general IHC practice). No RSRC1-specific dilution or background mechanism is supplied (HPA/UniProt evidence scope). |
| The IHC result lacks visibly discrete nuclear speckles. | HPA's approved speckle assignment comes from ICC-IF, while tissue IHC is described as nuclear and cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). | Assess the IHC section against its tissue-level nuclear and cytoplasmic pattern and cell identity; use the separate IF/ICC guide when discrete speckle imaging is the goal (HPA tissue IHC; HPA subcellular ICC-IF; general microscopy practice). |
Experiment evidence
IHC & IF images showing RSRC1 staining
A12655 · IHCImmunohistochemistry of RSRC1 in rat liver tissue with RSRC1 antibody at 5 μg/mL.
A12655 · IFImmunofluorescence of RSRC1 in rat liver tissue with RSRC1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000174891-RSRC1/tissue/cerebral+cortex#imid_11949127
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000174891-RSRC1/tissue/colon#imid_11948981
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000174891-RSRC1/tissue/liver#imid_11948990
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000174891-RSRC1/tissue/kidney#imid_11948991
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044792; green = RSRC1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044792; green = RSRC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RSRC1 IHC & IF
🧪Run duodenum first: its glandular cells must stain (HPA: High in duodenal glandular cells). Run adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the duodenum slide, score any morphologically distinct cells as internal negatives only after confirming that they lack specific staining above background.
Positive control tissue: Duodenum (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RSRC1 in A-431, U-251MG, U2OS, with annotated localisation: Nuclear speckles (approved) (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 RSRC1 knockout material or peptide blocking where suitable (standard IHC practice). For duodenum, quench endogenous peroxidase before chromogenic detection; block endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the selected A12655 rat-liver IHC caption also leaves its fixative unreported (A12655 tissue-IHC caption). The evidence does not establish whether frozen sections or IF are easier than paraffin IHC; ICC-IF offers a nuclear-speckle localisation reference (HPA: Nuclear speckles, approved). Check duodenal sections for endogenous peroxidase background when interpreting glandular staining (standard IHC practice).
HPA tissue IHC evidence for RSRC1
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 |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RSRC1 IHC Tips
Troubleshoot RSRC1 chromogenic IHC by checking retrieval, compartment-specific staining, cell identity, and controls before interpreting signal intensity.
What retrieval should I start with for RSRC1 in paraffin sections?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 minutes (page retrieval rule: nuclear antigen). Let sections cool in the retrieval buffer, then wash and compare staining with a section processed identically except for retrieval (standard IHC practice). Assess nuclear speckles as well as broader nuclear and cytoplasmic signal, since RSRC1 has these reported locations (HPA subcellular; UniProt Q96IZ7 localisation). If signal remains weak, compare a second retrieval condition on adjacent sections while holding antibody concentration and detection constant; record whether improved intensity also increases diffuse background (standard IHC practice).
How should I troubleshoot fixation-related loss of RSRC1 staining?
RSRC1-specific sensitivity to fixation is unknown from the supplied evidence; the rat liver image caption gives an antibody concentration but does not state a fixative (A12655 tissue-IHC caption). Document the specimen’s actual fixative, fixation interval, processing schedule, and section age before comparing cases (standard IHC practice). If a run is weak, stain a recently cut control section alongside the affected section using the same retrieval and detection steps (standard IHC practice). Interpret any improvement as evidence of a processing or storage difference, rather than proof of a particular RSRC1 fixation mechanism, and check whether expected nuclear staining is preserved (HPA tissue IHC; standard IHC practice).
Should RSRC1 staining be nuclear, cytoplasmic, or both?
Examine nuclear and cytoplasmic compartments separately: tissue IHC reports both, while cellular imaging places RSRC1 predominantly in nuclear speckles (HPA tissue IHC; HPA subcellular). RSRC1 can shuttle between nucleus and cytoplasm, so cytoplasmic signal alone is not automatically an artefact (UniProt Q96IZ7 localisation). At high magnification, confirm that puncta lie within counterstained nuclei and that cytoplasmic colour follows intact cell boundaries (standard IHC practice). If staining is exclusively membranous, reassess specificity and section artefacts, because RSRC1 has no annotated transmembrane segment and its reported locations are intracellular (UniProt Q96IZ7 topology and localisation).
Could isoform choice or epitope accessibility explain discordant RSRC1 staining?
Two RSRC1 isoforms are listed, but the supplied evidence does not map this antibody’s epitope or establish which isoforms it detects (UniProt Q96IZ7 isoforms; supplied antibody evidence). Do not interpret a negative section as isoform absence without an epitope map and an independent assay that distinguishes isoforms (standard IHC interpretation). Compare adjacent sections across retrieval conditions and inspect whether nuclear signal appears without a parallel rise in diffuse staining (standard IHC practice; HPA subcellular). RSRC1 has no annotated transmembrane segment or glycosylation sites, but those annotations do not establish epitope accessibility after paraffin processing (UniProt Q96IZ7 topology and glycosylation; standard IHC practice).
How can IF help check RSRC1 localisation alongside chromogenic IHC?
Use a separate IF or ICC optimisation for RSRC1, since the supplied A12655 image documents tissue IHC and does not establish an IF fixation or permeabilisation condition (A12655 tissue-IHC caption). Multiplex RSRC1 with a marker validated for the expected cell type in the specimen, then verify that the RSRC1 channel aligns with nuclei or nuclear speckles (HPA subcellular; standard IF practice). Choose spectrally separated fluorophores and inspect an unstained section to place the weaker channel away from prominent tissue autofluorescence (standard IF practice). Because RSRC1 is intracellular and lacks a transmembrane segment, optimise permeabilisation for nuclear access while checking that speckle detail is retained (UniProt Q96IZ7 topology; HPA subcellular; standard IF practice).
What controls help distinguish RSRC1 signal from chromogenic background?
Run a no-primary control through the same secondary reagent and chromogen steps to reveal detection-system background (standard IHC practice). Include a peroxidase block before peroxidase-based detection and compare staining before and after it if endogenous enzyme activity is suspected (standard chromogenic IHC practice). Titrate the primary antibody around a documented starting point; the supplied rat liver image reports 5 µg/mL, but that caption does not establish an optimum for other specimens (A12655 tissue-IHC caption). Judge improvements by retained intracellular staining and cleaner negative regions, including adipocytes reported as undetected in the tissue profile (HPA tissue IHC; UniProt Q96IZ7 localisation).
How should I quantify RSRC1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define cell types and nuclear versus cytoplasmic compartments before scoring, because RSRC1 is reported in both compartments in tissue and in nuclear speckles by cellular imaging (HPA tissue IHC; HPA subcellular). For each compartment, record the percentage of positive cells and an intensity-weighted H-score, using consistent thresholds across sections (standard IHC quantification). Normalise counts to the number of evaluable cells of the same type, or report positive-cell density per mm² of viable tissue when cell counts are impractical (standard IHC quantification). Exclude folds, tissue edges, and necrotic regions by a prespecified rule, and compare cases stained in the same run (standard IHC practice).
When is RSRC1 staining convincing rather than an artefact?
A convincing result shows reproducible intracellular staining in intact cells, with nuclear signal compatible with the reported nuclear and speckle locations and possible cytoplasmic signal (HPA tissue IHC; HPA subcellular; UniProt Q96IZ7 localisation). Use a reported high-staining cell population, such as duodenal glandular cells, as a reference while recognising that tissue IHC reliability is rated Supported with medium RNA–protein consistency (HPA tissue IHC). Treat exclusively membranous colour, staining confined to section edges or necrosis, and signal persisting in a no-primary control as reasons to investigate artefact (UniProt Q96IZ7 topology; standard IHC practice). Recheck peroxidase blocking and cell identity before assigning biological meaning to unexpected staining (standard chromogenic IHC practice).
Boster reagents
Best RSRC1 / Serine/Arginine-related protein 53 IHC Antibodies
A12655 has rat liver IHC and IF images (catalog image captions). Human, mouse and rat reactivity is listed; the pictured tissue is rat liver (catalog reactivity; image captions).
Real IHC data
Anti-RSRC1 Antibody
Cat # A12655
A12655 will render with its rat liver IHC image at 5 μg/mL (IHC image caption). Its IF image shows rat liver at 20 μg/mL; human, mouse and rat reactivity is listed (IF image caption; catalog reactivity).
Which to pick: For paraffin-section IHC, choose A12655: IHC-P is listed and its own IHC image shows rat liver at 5 μg/mL; the fixative is unreported (catalog applications; IHC image caption). For IF, A12655 has a rat liver image at 20 μg/mL; ICC validation is unreported (IF image caption; catalog applications). For work across species, A12655 lists human, mouse and rat reactivity, while the supplied IHC and IF images document rat liver only; its host is rabbit, and clonality is unreported (catalog reactivity; image captions; catalog host and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96IZ7 (RSRC1_HUMAN, Serine/Arginine-related protein 53).
- Human Protein Atlas. RSRC1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. RSRC1 subcellular location (ICC-IF): Localized to the nuclear speckles..
- Human Protein Atlas. RSRC1 antibody validation summary (1 antibodies).
- RSRC1 suppresses gastric cancer cell proliferation and migration by regulating PTEN expression. Molecular medicine reports 2019 — PMC6625388.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:15798186 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RSRC1 (Serine/Arginine-related protein 53). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A12655 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A12655); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-RSRC1, 5 μg/mL (datasheet A12655); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; verify antibody epitope coverage (UniProt); Regulation: Widely expressed; regulation unreported (UniProt). Evidence: UniProt Q96IZ7, HPA tissue IHC · Supported, HPA ICC-IF · nuclear speckles., Datasheet A12655, PMC6625388. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RSRC1/immunohistochemistry-ihc-guide.