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RPL28 / Large ribosomal subunit protein eL28 · IHC design guide
Design Immunohistochemistry for RPL28
Plan RPL28 chromogenic IHC on paraffin sections using the reported cytoplasmic tissue pattern (HPA tissue IHC). This guide covers fixation, cell-type controls and the A10323Y77 IHC dilution range of 1:50–1:200 (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPL28 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 | Cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | General cytoplasmic staining across multiple cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+3 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10323Y77) | |
| Caveat | Adipocytes lack detectable staining (HPA tissue IHC) | |
| Regulation | No specific regulator annotated (UniProt) | |
| Isoform / epitope | 5 isoforms; cytoplasmic target, epitope coverage unknown (UniProt) |
Section 1
Recommended RPL28 IHC & IF Protocols
The catalog antibody IHC-P protocol is accompanied by a published RPL28 protocol using paraffin-embedded tissue sections (PMC13553147 methods).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A10323Y77) |
| Fixation | Image fixative and duration unreported (datasheet A10323Y77); 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 citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane 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-RPL28, 1:50-1:200 (datasheet A10323Y77) |
| 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 | RPL28-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 citrate pH 6.0 heat retrieval at 95–98 °C for 20 min (page antigen retrieval); optimize with tissue controls.
Section 2
What Is the Expected RPL28 Staining Pattern?
RPL28 is a cytoplasmic component of the large ribosomal subunit and has no transmembrane segment (UniProt P46779). In paraffin section IHC, expect general cytoplasmic staining, including high staining in appendix and colon glandular cells, bone marrow hematopoietic cells, and cerebellar Purkinje cells (HPA: tissue IHC). HPA rates the tissue pattern Approved, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC).
| Clear cytoplasmic staining in appendix or colon glandular cells, with tissue structures remaining easy to distinguish. | This matches the reported general cytoplasmic pattern and High staining in those cell populations (HPA: tissue IHC). Evaluate staining in identified cells; a positive area alone does not establish which cells carry the signal (general IHC practice). |
| Strong nuclear only staining, or a sharply membrane confined outline, with little cytoplasmic signal. | That distribution conflicts with the reported cytoplasmic IHC pattern (HPA: tissue IHC) and cytoplasmic, nontransmembrane protein annotation (UniProt P46779). Treat it as a possible artefact and check controls before assigning it to RPL28 (general IHC practice). |
| Prominent staining in adipocytes, ovarian follicle cells, smooth muscle cells, or chondrocytes. | HPA lists these specific cell populations as Not detected, so staining there warrants scrutiny; the designation is cell specific, not a claim that the whole tissue must be blank (HPA: tissue IHC). Cross reactivity or endogenous detection activity are possibilities to investigate with controls (general IHC practice). |
| Diffuse color covers cells and surrounding tissue, obscuring cytoplasmic boundaries. | A uniform deposit is difficult to reconcile with an interpretable cellular pattern (general IHC practice); HPA describes RPL28 staining as generally cytoplasmic (HPA: tissue IHC). Compare a no primary control and review blocking, washes, and chromogen development (general IHC practice). |
| No detectable signal in the expected cells of an appendix or colon section. | HPA reports High staining in glandular cells of both tissues, making a blank result a reason to examine assay performance (HPA: tissue IHC). Confirm that the expected cells are present, then review the antibody and detection workflow alongside a positive control (general IHC practice). |
💡Expected RPL28 appearanceA convincing positive is distinguishable cytoplasmic staining in HPA High cell populations, such as appendix or colon glandular cells (HPA: tissue IHC), whereas isolated nuclear or membrane confined color warrants artefact checks (UniProt P46779; general IHC practice).
| Cell population and tissue | HPA reports High staining in several defined populations but Not detected in adipocytes, follicle cells, smooth muscle cells, and chondrocytes; select and score controls by cell type (HPA: tissue IHC; general IHC practice). |
| Evidence strength | The tissue profile is Approved with medium staining to RNA consistency, and HPA050459 is Approved for IHC; these labels support use of the observed pattern while leaving ambiguous staining to be checked with controls (HPA: tissue IHC; HPA: HPA050459 validation). |
| Subcellular context | UniProt places RPL28 in the cytoplasm as a large ribosomal subunit component with no transmembrane segment; HPA describes IHC as generally cytoplasmic (UniProt P46779; HPA: tissue IHC). |
| Target specific fixation and retrieval response | No target specific fixation sensitivity or antigen retrieval condition is established by the supplied UniProt and HPA records; determine workable IHC conditions with assay controls (UniProt P46779; HPA: tissue IHC; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected glandular cells are blank in appendix or colon. | These cells are reported High, so absence may reflect an assay or specimen problem; the pattern alone cannot identify which step failed (HPA: tissue IHC; general IHC practice). | Check tissue identity and cell preservation, then compare a working positive control; review retrieval, primary antibody application, and detection conditions as general IHC workflow checks (general IHC practice). |
| Color appears chiefly in nuclei or along cell membranes. | This differs from HPA's general cytoplasmic IHC pattern and UniProt's cytoplasmic, nontransmembrane annotation (HPA: tissue IHC; UniProt P46779). | Confirm the compartment with the counterstain and cell morphology, then inspect a no primary control and detection background before calling the signal specific (general IHC practice). |
| Cells reported Not detected show conspicuous staining. | HPA reports no detection in the specified adipocytes, follicle cells, smooth muscle cells, and chondrocytes; off target binding or endogenous activity could explain unexpected color (HPA: tissue IHC; general IHC practice). | Score the named cell type, compare an expected positive population, and use no primary and appropriate detection controls to investigate background (general IHC practice). |
| Diffuse background prevents cell level scoring. | Excess nonspecific binding or detection background can mask a cellular pattern (general IHC practice); the reported IHC distribution is generally cytoplasmic (HPA: tissue IHC). | Compare a no primary control, then review blocking, wash steps, detection reagent exposure, and chromogen development within the established IHC workflow (general IHC practice). |
| Weak staining in thyroid, parathyroid, lung alveolar, or placental trophoblastic cells is being called an assay failure. | HPA lists these cell populations as Low, so faint signal there is less informative about assay performance than the listed High populations (HPA: tissue IHC). | Judge the run against an expected High cell population and score each tissue's named cell type separately (HPA: tissue IHC; general IHC practice). |
| IF/ICC: what distribution should be compared with an image? | HPA reports supported endoplasmic reticulum and cytosol localization in ICC-IF, while its tissue IHC summary says general cytoplasmic expression (HPA: subcellular ICC-IF; HPA: tissue IHC). | Use the ER and cytosol annotation to interpret IF/ICC localization; evaluate paraffin section IHC against the tissue IHC pattern on this page (HPA: subcellular ICC-IF; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing RPL28 staining
A10323Y77 · IHCImmunohistochemistry (IHC) analyzes of RPL28 (Y77) pAb in paraffin-embedded human breast carcinoma tissue at 1:100.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000108107-RPL28/tissue/cerebral+cortex#imid_15054446
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000108107-RPL28/tissue/colon#imid_15054380
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000108107-RPL28/tissue/liver#imid_15054373
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000108107-RPL28/tissue/kidney#imid_15054374
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA050459; green = RPL28, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA050459; green = RPL28, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPL28 IHC & IF
🧪Run appendix first and score its glandular cells, where RPL28 staining is High (HPA: High in appendix glandular cells); use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes). On the appendix slide, cells outside the glandular compartment should be compared with control-level background, but the supplied HPA rows do not establish an internal-negative cell type.
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 RPL28 in HEK293, MCF-7, U2OS, with annotated localisation: Endoplasmic reticulum (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and immunoglobulin-class-matched isotype controls appropriate to the primary antibody’s clonality; use RPL28 knockout tissue as a biological negative if available. For appendix chromogenic IHC, quench endogenous peroxidase and check for endogenous biotin background if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A10323Y77 paraffin-section caption does not state the fixative (caption: paraffin-embedded human breast carcinoma tissue at 1:100; fixative unreported). Retrieval dependency is unreported; optimize antigen retrieval empirically for paraffin IHC (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier; in appendix, endogenous peroxidase background can complicate chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for RPL28
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 → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Chondrocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPL28 IHC Tips
Use cytoplasmic staining, cell identity and matched controls to troubleshoot RPL28 chromogenic IHC in paraffin sections (HPA tissue IHC; selected image caption).
Which retrieval should I start with for weak RPL28 staining in paraffin sections?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule: cytoplasmic antigen). Let sections cool in buffer, then compare the same antibody dilution and chromogen development across runs so retrieval is the main variable (standard IHC practice). If staining remains weak, test a stronger retrieval condition on a serial section while checking whether tissue morphology and background deteriorate (standard IHC practice). Judge improvement by cytoplasmic signal in glandular cells of colon or appendix, which show high staining in the reference profile (HPA tissue IHC).
Could fixation explain loss of RPL28 signal in my paraffin sections?
The selected image documents paraffin-embedded human breast carcinoma at 1:100, but does not report its fixative (selected image caption: A10323Y77). No target-specific fixation sensitivity is supplied, so a weak result cannot be assigned to a particular fixative or fixation duration (evidence supplied for RPL28). Record the actual fixative and processing history, then compare serial sections from matched material while holding retrieval, antibody dilution and detection constant (standard IHC practice). Include a known staining-positive section in each run; colon glandular cells are a documented high-staining reference for assessing whether the run worked (HPA tissue IHC).
What staining pattern should count as plausible RPL28 signal?
Expect predominantly cytoplasmic chromogenic staining because RPL28 is annotated in the cytoplasm and tissue IHC shows general cytoplasmic expression (UniProt P46779; HPA tissue IHC). Supported IF locations include cytosol and endoplasmic reticulum, which can make staining look diffuse or finely granular at light-microscope resolution (HPA subcellular). RPL28 has no annotated transmembrane segment, so a sharply membrane-restricted outline needs independent scrutiny (UniProt P46779 topology). Compare suspicious staining with cell boundaries and a counterstain on the same section, then inspect a no-primary control for detection artefact (standard IHC practice). Do not score isolated nuclear deposits as convincing signal without further validation (UniProt P46779 localisation; standard IHC practice).
How should I troubleshoot an unexpected pattern if the antibody epitope is unknown?
RPL28 has 5 annotated isoforms, but the supplied product evidence does not identify the antibody epitope or establish isoform coverage (UniProt P46779 isoforms; selected image caption). It also lists N-acetylserine at residue 2 and phosphoserine at residue 115; neither annotation shows that this antibody recognizes a modified or unmodified state (UniProt P46779). Request the immunogen or epitope specification before attributing an uneven IHC pattern to splice variation or a modification (standard antibody validation practice). In the meantime, compare serial sections under identical retrieval and detection settings, and score only reproducible cytoplasmic staining in identifiable cells (HPA tissue IHC; standard IHC practice).
How can IF help assess the cell source of RPL28 IHC staining?
Use IF as a separate localization check: HPA reports supported RPL28 signal in cytosol and endoplasmic reticulum, but the selected catalog image documents tissue IHC only (HPA subcellular; selected image caption). For breast carcinoma material, multiplex with an epithelial marker to identify the cells contributing signal in the IHC field (selected image caption; standard IF practice). Choose a far-red fluorophore if the specimen has strong shorter-wavelength autofluorescence, and inspect single-label and no-primary controls (standard IF practice). For intracellular access, test mild permeabilisation; RPL28 lacks a transmembrane segment, while the supplied evidence does not establish the antibody epitope's membrane sidedness (UniProt P46779 topology; standard IF practice).
What should I check when RPL28 staining appears widespread or muddy?
Widespread cytoplasmic signal is plausible for RPL28, so judge background against cell morphology and controls instead of assuming every stained cell is false positive (HPA tissue IHC). Run a no-primary control, block endogenous peroxidase before chromogenic detection, and adjust protein blocking or washes if nonspecific deposition persists (standard chromogenic IHC practice). Compare the catalog image's 1:100 dilution with a small dilution series on matched sections, keeping retrieval and DAB development fixed (selected image caption: A10323Y77; standard IHC practice). If signal coats edges, folds or damaged regions while intact cell cytoplasm is indistinct, exclude those regions from scoring (standard IHC practice).
How should I quantify variable RPL28 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis area before scoring, and exclude folds, necrosis and section edges using the counterstain (standard IHC quantification practice). For intact cells, record the percentage positive and intensity on a fixed 0–3 scale, then calculate an H-score from intensity-weighted percentages (standard IHC quantification practice). Normalize comparisons to the number of eligible cells or assessed tissue area, and keep section thickness, retrieval, dilution and chromogen development consistent (standard IHC practice). Report cell types separately: the reference profile shows high staining in colon glandular cells but no detected staining in adipocytes, so pooled tissue averages can obscure composition (HPA tissue IHC).
How do I distinguish convincing RPL28 signal from staining artefact?
Convincing signal should be reproducible in intact cell cytoplasm, consistent with the annotated cytoplasmic location and general tissue-IHC profile (UniProt P46779; HPA tissue IHC). Cell identity matters: high staining is reported in appendix glandular cells, whereas adipocytes are listed as not detected in the reference profile (HPA tissue IHC). Treat isolated nuclear staining, edge-heavy deposits and staining confined to necrotic regions as suspect, and inspect a no-primary control for endogenous enzyme or detection signal (UniProt P46779 localisation; standard IHC practice). Interpret a negative or weak field cautiously because the reference tissue-IHC assessment is Approved with medium consistency against RNA expression (HPA tissue IHC).
Boster reagents
Best RPL28 / Large ribosomal subunit protein eL28 IHC Antibodies
Catalog anti-RPL28 antibodies list human, mouse and rat reactivity; real IHC imagery shows paraffin-embedded human breast carcinoma tissue (catalog; A10323Y77 image caption).
Real IHC data
Anti-RPL28 (Y77) Antibody
Cat # A10323Y77
A10323Y77 is the only card that will render, with an IHC image of paraffin-embedded human breast carcinoma tissue at 1:100 (A10323Y77 image caption). Its listed applications are IP, IHC and WB, and its listed reactivity is human, mouse and rat (catalog).
Which to pick: For tissue IHC, choose A10323Y77 because its own image documents paraffin-section staining; the fixative is unreported (A10323Y77 image caption). For IF/ICC, A10323-1 lists IF at 1:50, but has no IF figure and does not list ICC (catalog). Both list human, mouse and rat reactivity and a rabbit host; neither reports a clone, and the shown tissue IHC evidence is human only (catalog; A10323Y77 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P46779 (RL28_HUMAN, Large ribosomal subunit protein eL28).
- Human Protein Atlas. RPL28 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RPL28 subcellular location (ICC-IF): Localized to the endoplasmic reticulum and cytosol..
- Human Protein Atlas. RPL28 antibody validation summary (1 antibodies).
- Knockdown of RPL28 Inhibits the Progression of Clear Cell Renal Cell Carcinoma. Cancer reports (Hoboken, N.J.) 2026 — PMC13553147.
- scRNA-seq deciphers molecular mechanisms of endocrine disruptor 4-nonylphenol impairing spermatogenesis in mice. Cell biology and toxicology 2025 — PMC12494639.
- The Ribosomal Protein L28 Gene Induces Sorafenib Resistance in Hepatocellular Carcinoma. Frontiers in oncology 2021 — PMC8299949.
- PubMed PMID:7772601 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15057824 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPL28 (Large ribosomal subunit protein eL28). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A10323Y77 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A10323Y77); 2. Fixation: Image fixative and duration unreported (datasheet A10323Y77); 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 citrate buffer, pH 6.0 , 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-RPL28, 1:50-1:200 (datasheet A10323Y77); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10323Y77); Caveat: Adipocytes lack detectable staining (HPA tissue IHC); Isoform / epitope: 5 isoforms; cytoplasmic target, epitope coverage unknown (UniProt); Regulation: No specific regulator annotated (UniProt). Evidence: UniProt P46779, HPA tissue IHC · Approved, HPA ICC-IF · endoplasmic reticulum and cytosol., Datasheet A10323Y77, PMC13553147. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPL28/immunohistochemistry-ihc-guide.