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RPS7 / Small ribosomal subunit protein eS7 · IHC design guide
Design Immunohistochemistry for RPS7
RPS7 shows general cytoplasmic staining in tissue IHC (HPA tissue IHC). This guide uses the catalog antibody’s illustrated human breast staining at 1:100 as a starting point for paraffin-section planning (datasheet: A04736).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPS7 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 | General cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in glandular and epithelial cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show only medium consistency (HPA tissue IHC) | |
| Regulation | No stimulus-linked regulation annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–194 chain (UniProt) |
Section 1
Recommended RPS7 IHC & IF Protocols
The catalog antibody protocol is paired with published RPS7 IHC methods for colorectal cancer tissue microarrays and paraffin sections (PMC4868722; PMC9584673).
| Sample | Paraffin-embedded rat brain tissue; fixative not specified (datasheet A04736) |
| Fixation | Image fixative and duration unreported (datasheet A04736); 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-RPS7, 1:50-1:200 (datasheet A04736) |
| 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 | RPS7-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. 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: nuclear antigen).
Section 2
What Is the Expected RPS7 Staining Pattern?
RPS7 is a small ribosomal subunit protein found in the cytoplasm and nucleolus, with centrosomal localization also reported; it has no transmembrane segment (UniProt P62081). In paraffin tissue sections, expect predominantly cytoplasmic staining across many cell types (HPA: general cytoplasmic expression; low tissue specificity). HPA rates its tissue IHC evidence Approved, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC).
| Cytoplasmic staining is strong in appendix glandular cells and bone marrow hematopoietic cells (HPA: High in both). | This fits the reported IHC pattern and gives useful positive tissue checks (HPA: general cytoplasmic expression; High in both). Compare cell types within each section, since HPA reports tissue staining with medium consistency against RNA expression (HPA: tissue IHC). |
| Signal is confined to cell borders or another sharply membrane-like pattern. | Treat a predominantly membrane-bound result as suspect: RPS7 has no transmembrane segment, while HPA describes general cytoplasmic IHC staining (UniProt P62081 topology; HPA: tissue IHC). Nuclear or nucleolar staining alone is less decisive because UniProt also places RPS7 in the nucleolus (UniProt P62081). |
| Adipocytes or smooth muscle cells stain strongly and reproducibly. | These are discordant with the supplied HPA examples, where both cell types are Not detected (HPA: adipocytes and smooth muscle cells). Check for antibody cross-reactivity or endogenous chromogenic activity before calling them RPS7-positive (general IHC practice). Do not treat either example as a universal negative control (HPA: low tissue specificity; medium consistency). |
| Brown signal covers tissue, extracellular spaces, or the negative control without clear cellular boundaries. | A widespread deposit is background rather than a readable RPS7 localization pattern (general IHC practice; HPA: general cytoplasmic expression). Review blocking, washes, detection reagent, and chromogen development alongside a control that omits primary antibody (general IHC practice). |
| No staining appears in appendix glandular cells or bone marrow hematopoietic cells (HPA: High in both). | An absent signal in these reported high-staining cells makes the run difficult to interpret (HPA: High in both). First check section integrity, positive control, antibody and detection steps, and counterstain visibility (general IHC practice). HPA's Approved rating does not establish fixation sensitivity or guarantee every specimen will stain (HPA: tissue IHC). |
💡Expected RPS7 appearanceCall a result consistent with RPS7 when cytoplasmic staining is clear in reported high-staining cell types, such as appendix glandular cells, while widespread noncellular deposit or strong staining in reported Not detected cells prompts investigation (HPA: general cytoplasmic expression; High in appendix glandular cells; Not detected in adipocytes and smooth muscle cells; general IHC practice).
| Compartment and topology | RPS7 is cytoplasmic and nucleolar, with centrosomal localization reported and no transmembrane segment (UniProt P62081). HPA's tissue IHC profile is generally cytoplasmic (HPA: tissue IHC). Assess the dominant cellular pattern rather than rejecting every nuclear focus (UniProt P62081; HPA: tissue IHC). |
| Cell type and tissue context | HPA reports High staining in several epithelial, hematopoietic, neural, and glial cell groups; prostate glandular cells and skeletal muscle myocytes are Low, while adipocytes and smooth muscle cells are Not detected in the supplied examples (HPA: tissue IHC). Choose comparisons by cell type, not tissue name alone (general IHC practice). |
| Strength of validation | The listed antibody HPA056586 is Approved for IHC, and the tissue profile has medium consistency with RNA expression (HPA: antibody validation; tissue IHC). An IHC Enhanced designation is not supplied (HPA: antibody validation). Interpret an unexpected compartment or cell type with controls rather than assuming the pattern is confirmed independently (general IHC practice). |
| Epitope and fixation sensitivity | The supplied record gives no antibody epitope or target-specific fixation effect (UniProt P62081; HPA: supplied records). Antigen retrieval can be checked against the catalog antibody's IHC instructions as a general paraffin IHC workflow step; do not infer an RPS7-specific retrieval requirement from HPA staining levels (general IHC practice; HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known high-staining cells show no chromogenic signal (HPA: High in appendix glandular cells and bone marrow hematopoietic cells). | The run may have a detection, reagent, retrieval, or section problem (general IHC practice); the supplied sources identify no RPS7-specific fixation failure mode (UniProt P62081; HPA: supplied records). | Inspect a positive control and the catalog antibody's IHC instructions; verify reagent sequence, antigen retrieval, and chromogen activity (general IHC practice). |
| The entire section is brown, including spaces outside cells. | Nonspecific reagent binding or excess chromogen development can obscure cellular localization (general IHC practice). | Check the primary-omission control, blocking and washes, and development time; score RPS7 only where cells show interpretable staining (general IHC practice; HPA: general cytoplasmic expression). |
| Signal appears in adipocytes or smooth muscle cells despite the supplied HPA Not detected examples (HPA: tissue IHC). | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA's medium RNA-staining consistency also limits a categorical biological conclusion (HPA: tissue IHC). | Compare primary-omission and detection controls, then assess the cell boundaries and cytoplasmic pattern before assigning positivity (general IHC practice; HPA: general cytoplasmic expression). |
| Staining is restricted to membranes or has no recognizable cytoplasmic component. | That distribution conflicts with HPA's general cytoplasmic IHC profile and RPS7's lack of a transmembrane segment (HPA: tissue IHC; UniProt P62081 topology). | Recheck the negative control and tissue morphology; repeat with the IHC-validated antibody's documented conditions if the localization remains discordant (general IHC practice; HPA: antibody IHC Approved). |
| A section shows mixed cytoplasmic and nuclear or nucleolar signal. | RPS7 has reported cytoplasmic and nucleolar locations, so a nuclear component is biologically plausible (UniProt P62081); HPA summarizes tissue IHC as generally cytoplasmic (HPA: tissue IHC). | Score cytoplasmic and nuclear components separately, verify cellular morphology, and use controls before treating an unusually dominant nuclear pattern as specific (general IHC practice). |
| What should an IF/ICC comparison show? | HPA reports endoplasmic reticulum localization as Approved and cytosol localization as Supported in ICC-IF; these are modality-specific observations (HPA: subcellular ICC-IF). | For an IF/ICC comparison, look for cellular signal compatible with those compartments; HPA lists images from A-431, U-251MG, and U2OS cells (HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing RPS7 staining
A04736 · IHCImmunohistochemistry of paraffin-embedded rat brain using RPS7 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 mM
A04736 · IHCImmunohistochemistry of paraffin-embedded human breast using RPS7 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10
M04736 · IFImmunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical epithelial adenocarcinoma cell
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000171863-RPS7/tissue/cerebral+cortex#imid_16308435
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000171863-RPS7/tissue/colon#imid_16308361
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000171863-RPS7/tissue/liver#imid_16308362
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000171863-RPS7/tissue/kidney#imid_16308363
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA056586; green = RPS7, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA056586; green = RPS7, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPS7 IHC & IF
🧪Run appendix first: glandular cells should stain (HPA: High in appendix glandular cells). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); no internal-negative cell population is identified for the appendix slide, so cells showing only counterstain should not be called biologically RPS7-negative without validation.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPS7 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (approved), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and clonality-matched isotype controls, plus an RPS7 knockout sample or validated peptide-block control (standard IHC practice). Block endogenous peroxidase and inspect inflammatory cells for background staining in appendix sections (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04736 paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). The caption uses microwave retrieval in 10 mM PBS, pH 7.2, at 1:100 for rat brain; it does not establish that retrieval condition for appendix (selected-SKU tissue-IHC caption). Whether frozen sections or IF are easier is unreported; appendix inflammatory cells can contribute endogenous-peroxidase background (standard IHC practice).
HPA tissue IHC evidence for RPS7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 → |
| Bone marrow | Hematopoietic 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 → |
Section 3
Advanced RPS7 IHC Tips
Troubleshoot RPS7 chromogenic IHC by checking retrieval, compartment pattern, controls and cell level scoring before interpreting changes in staining.
How should I retrieve RPS7 in paraffin sections when staining is weak?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting: nuclear antigen). Let sections cool in retrieval buffer, then compare staining and tissue integrity with an untreated section (standard IHC practice). If staining remains weak, test microwave retrieval in 10 mM PBS at pH 7.2, the condition reported for paraffin-embedded rat brain with antibody A04736 (A04736 tissue-IHC caption). Hold antibody dilution and detection conditions constant during that comparison so a change in cytoplasmic or nucleolar signal can be attributed to retrieval (standard IHC practice; UniProt P62081 localisation).
Could fixation explain weak or uneven RPS7 staining?
Target-specific fixation sensitivity is unknown because no RPS7 fixation comparison is supplied (provided RPS7 evidence). The A04736 image identifies paraffin-embedded rat brain but does not state its fixative, so it cannot establish a fixation condition (A04736 tissue-IHC caption). Record fixative, fixation duration and processing history for each specimen, and compare sections processed together when investigating uneven staining (standard IHC practice). Use a consistent control section across runs, then assess whether retrieval or detection changes restore signal without increasing background (standard IHC practice). Do not assign a fixation effect from RPS7 localisation or acetylation alone (UniProt P62081 localisation and modified residues).
Which RPS7 staining compartments should I accept in tissue sections?
Expect predominantly cytoplasmic tissue staining, while assessing discrete nucleolar signal separately (HPA tissue IHC: general cytoplasmic expression; UniProt P62081 localisation). RPS7 participates in the cytoplasmic small ribosomal subunit, and subunit assembly occurs in the nucleus (UniProt P62081 function and localisation). HPA cell imaging also reports endoplasmic reticulum and cytosol localisation, but that cell imaging pattern does not establish a required compartment pattern in every tissue section (HPA subcellular). Score cytoplasmic and nucleolar staining independently, with matched morphology and a negative reagent control, especially when diffuse nuclear colour could reflect background (standard IHC practice). Treat isolated cell-surface staining cautiously because RPS7 has no annotated transmembrane segment (UniProt P62081 topology).
How should I troubleshoot an unexpected RPS7 epitope pattern?
RPS7 has one annotated 194-residue chain and no annotated isoforms or transmembrane segment (UniProt P62081 processing, isoforms and topology). Its listed modifications include N-terminal acetylation and lysine acetylation, including an alternate site at residue 74; the supplied evidence does not map the antibody epitope (UniProt P62081 modified residues; provided antibody evidence). If staining changes after retrieval, compare adjacent sections under controlled retrieval conditions before attributing the difference to an epitope or modification (standard IHC practice). Record the antibody identifier and detection conditions, and seek epitope documentation before claiming that a compartment-specific signal represents a distinct RPS7 form (standard IHC practice; UniProt P62081 isoforms).
How can IF help assess an ambiguous RPS7 IHC pattern?
Use IF as a complementary localisation check, while keeping the chromogenic IHC result as the tissue readout (standard IHC/IF practice). Multiplex RPS7 with a marker for the cell population being assessed, and examine whether signal shares those cells and the expected cytosolic or nucleolar compartment (HPA tissue IHC: general cytoplasmic expression; UniProt P62081 localisation). Choose spectrally separated fluorophores and a channel with low tissue autofluorescence, checking an unstained section before interpreting overlap (standard IF practice). RPS7 has no transmembrane segment, so optimise permeabilisation for intracellular epitopes after fixation; the IHC caption supplies no IF permeabilisation condition for A04736 (UniProt P62081 topology; A04736 tissue-IHC caption).
What should I check when RPS7 chromogenic staining is widespread?
Widespread cytoplasmic staining can be plausible because HPA reports general cytoplasmic RPS7 expression, so judge background against cell morphology and controls (HPA tissue IHC; standard IHC practice). Include a no-primary control and inspect edge staining, folds and necrotic areas before increasing antibody concentration (standard IHC practice). For peroxidase detection, use an endogenous peroxidase block and check whether residual colour appears in the no-primary control (standard chromogenic IHC practice). Titrate around the reported 1:100 dilution only for the A04736 paraffin rat-brain example, while keeping retrieval and development time fixed during the comparison (A04736 tissue-IHC caption; standard IHC practice).
How should I quantify RPS7 across comparable tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then quantify the percentage of positive cells and staining intensity with a consistent threshold (standard IHC practice). An H-score combines intensity categories with their positive-cell percentages; record cytoplasmic and nucleolar scores separately because both compartments are biologically plausible for RPS7 (standard IHC practice; UniProt P62081 localisation). For spatial comparisons, report positive-cell density per mm² of viable tissue and normalise counts to the corresponding area or total eligible cells (standard image-analysis practice). Analyse matched section thickness, retrieval, dilution and chromogen development, and exclude folds, edges and necrosis by the same prespecified rule (standard IHC practice).
When is an apparent RPS7-positive cell likely to be an artefact?
A credible positive follows intact cell morphology and a cytoplasmic pattern, with possible nucleolar signal consistent with RPS7 biology (HPA tissue IHC: general cytoplasmic expression; UniProt P62081 localisation). High staining has been reported in appendix glandular cells and bone-marrow hematopoietic cells, whereas adipocytes and smooth muscle cells were reported as not detected (HPA tissue IHC). Treat isolated membrane rims, section edges, folds and necrotic deposits as suspect until they persist in well-preserved regions and appropriate controls (UniProt P62081 topology; standard IHC practice). If a no-primary control develops colour, investigate endogenous enzyme activity or detection background before interpreting a change in RPS7 expression (standard chromogenic IHC practice).
Boster reagents
Best RPS7 / Small ribosomal subunit protein eS7 IHC Antibodies
Validated anti-RPS7 data cover paraffin-section IHC in rat brain, human breast, and mouse spinal cord (A04736 image captions), and IF in human HeLa cells (M04736 image caption).
Real IHC data
Anti-40S ribosomal protein S7 RPS7 Antibody
Cat # A04736
Real IF data
Anti-RPS7 Antibody (C-Term)
Cat # M04736
A04736 lists IHC and ICC/IF for human, mouse, and rat (A04736 applications and reactivity); its IHC images show paraffin-embedded rat brain, human breast, and mouse spinal cord (A04736 image captions). M04736 lists IF for human, mouse, and rat (M04736 applications and reactivity), with an IF image of paraformaldehyde-fixed, permeabilized HeLa cells (M04736 image caption).
Which to pick: Choose A04736 for paraffin-section tissue IHC (A04736 applications and image captions); its IHC captions do not report the fixative (A04736 image captions). Choose M04736 for cell IF (M04736 application and image caption), or A04736 when ICC is required (A04736 applications). Both list human, mouse, and rat reactivity, but only A04736 lists IHC (A04736 and M04736 applications and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P62081 (RS7_HUMAN, Small ribosomal subunit protein eS7).
- Human Protein Atlas. RPS7 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RPS7 subcellular location (ICC-IF): Localized to the endoplasmic reticulum and cytosol..
- Human Protein Atlas. RPS7 antibody validation summary (1 antibodies).
- RPS7 inhibits colorectal cancer growth via decreasing HIF-1α-mediated glycolysis. Oncotarget 2016 — PMC4868722.
- Identification of RPS7 as the Biomarker of Ferroptosis in Acute Kidney Injury. BioMed research international 2022 — PMC9584673.
- Mutation of the diamond-blackfan anemia gene Rps7 in mouse results in morphological and neuroanatomical phenotypes. PLoS genetics 2013 — PMC3561062.
- Small ribosomal protein subunit S7 suppresses ovarian tumorigenesis through regulation of the PI3K/AKT and MAPK pathways. PloS one 2013 — PMC3823983.
- PubMed PMID:8522193 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPS7 (Small ribosomal subunit protein eS7). Expected localisation General 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: A04736 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat brain tissue; fixative not specified (datasheet A04736); 2. Fixation: Image fixative and duration unreported (datasheet A04736); 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-RPS7, 1:50-1:200 (datasheet A04736); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific); Caveat: Staining and RNA show only medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–194 chain (UniProt); Regulation: No stimulus-linked regulation annotated (UniProt). Evidence: UniProt P62081, HPA tissue IHC · Approved, HPA ICC-IF · endoplasmic reticulum and cytosol., Datasheet A04736, PMC4868722, PMC9584673. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPS7/immunohistochemistry-ihc-guide.