RRBP1 / Ribosome-binding protein 1 · IHC design guide

Design Immunohistochemistry for RRBP1

Plan RRBP1 chromogenic IHC in paraffin sections using the cytoplasmic tissue pattern as a reference (HPA tissue IHC). Start with the IHC-validated antibody A07074-1 at 0.5–1 µg/mL (datasheet A07074-1), and compare samples prepared with consistent fixation.

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for RRBP1 (IHC for RRBP1): expected localisation Cytoplasmic tissue staining (HPA tissue IHC); ER membrane (UniProt), antibody A07074-1, validated IHC image, and IHC protocol steps
Printable RRBP1 IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC); ER membrane (UniProt), antibody A07074-1, controls and protocol steps. Open the full RRBP1 IHC guide →

RRBP1 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); ER membrane (UniProt)
Staining pattern Cytoplasmic across tissues; high in gut endocrine cells (HPA tissue IHC)
Antigen retrieval Citrate pH 6 HIER, heat-mediated (datasheet A07074-1)
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 paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific)
Caveat Staining intensity varies by cell type (HPA tissue IHC)
Regulation No expression regulator annotated (UniProt)
Isoform / epitope 2 isoforms; epitope differences unspecified (UniProt)
Section 1

Recommended RRBP1 IHC & IF Protocols

The catalog antibody has a datasheet IHC-P protocol (datasheet: A07074-1). Three published RRBP1 IHC protocols provide tissue-specific examples (PMC6646231; PMC6347782; PMC3439379).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human intestinal cancer tissue; fixative not specified (datasheet A07074-1)
FixationImage fixative and duration unreported (datasheet A07074-1); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: Citrate pH 6, 20 min (datasheet A07074-1)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A07074-1)
Primary antibodyRabbit anti-RRBP1, 0.5-1μg/ml (datasheet A07074-1)
Primary incubationOvernight at 4 °C (datasheet A07074-1)
DetectionStreptavidin-biotin complex (SABC), DAB chromogen (datasheet A07074-1)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultRRBP1-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in all tissues. No signal in the no-primary control.
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 for the catalog antibody (datasheet: A07074-1); published protocols also used citrate or EDTA (PMC6347782; PMC6646231).
Section 2

What Is the Expected RRBP1 Staining Pattern?

RRBP1 is an endoplasmic reticulum membrane protein with a short lumenal segment and a large cytoplasmic region (UniProt Q9P2E9 topology). Expect cytoplasmic staining across tissues, with high staining in selected cell populations such as bronchial respiratory epithelium and cerebellar Purkinje cells (HPA tissue IHC: Enhanced; high in those cells). HPA reports high consistency between antibody staining and RNA expression (HPA tissue IHC: reliability description).

What am I looking at on my slide?
Cytoplasmic chromogen in bronchial respiratory epithelial cells or cerebellar Purkinje cells.This fits the reported compartment and high staining in those cells (HPA tissue IHC: cytoplasmic expression; high in respiratory epithelial and Purkinje cells). Judge the labeled cells against nearby tissue structure, since the HPA profile describes expression across tissues rather than a single positive cell type (HPA tissue IHC: cytoplasmic expression in all tissues).
Predominantly nuclear staining, with little cytoplasmic signal.Treat this as a compartment mismatch: UniProt places RRBP1 at the ER membrane, while HPA reports cytoplasmic tissue staining (UniProt Q9P2E9: subcellular location; HPA tissue IHC: cytoplasmic expression). Review the counterstain and detection controls before assigning the nuclear signal to RRBP1 (general IHC practice).
Strong color in an unexpected cell population, especially without the expected positive cells staining.The distribution may reflect cross-reactivity or endogenous detection activity; appearance alone cannot distinguish them (general IHC practice). Compare cell identity with HPA's cell-specific observations and check appropriate detection controls (HPA tissue IHC: positive cell list; general IHC practice).
A diffuse wash of color obscures cell boundaries or extends through cell-free areas.This is background rather than a convincing cellular result (general IHC practice). Check blocking, washes, chromogen development and detection controls; the HPA observation is cytoplasmic staining within cells (general IHC practice; HPA tissue IHC: cytoplasmic expression).
No detectable signal in a section containing a reported high-staining cell population.First confirm that the relevant cells are present and the stain run worked (general IHC practice). Absence in, for example, bronchial respiratory epithelium conflicts with its reported high staining, but one failed section does not establish absent RRBP1 expression (HPA tissue IHC: high in bronchial respiratory epithelial cells).
💡Expected RRBP1 appearanceCall a result positive when chromogen is cellular and predominantly cytoplasmic in the identified cell population, with strong signal in an HPA-reported high-staining population; isolated nuclear or cell-free color is suspect (HPA tissue IHC: cytoplasmic expression and high-staining cell list; UniProt Q9P2E9: ER membrane; general IHC practice).
How each factor affects the staining
Compartment and topologyRRBP1 has one transmembrane segment at residues 8–28 and a cytoplasmic region at 29–1410 (UniProt Q9P2E9 topology). This supports a cytoplasmic, ER-associated interpretation; it does not identify the epitope recognized by a particular antibody (UniProt Q9P2E9 topology).
Cell population and tissueHPA reports high staining in several defined populations, but low staining in breast adipocytes, cardiomyocytes and skeletal myocytes (HPA tissue IHC: positive and low lists). Score the relevant cell type instead of treating every cell in a section as an equally strong control (general IHC practice).
Evidence behind the reference patternHPA rates the tissue IHC profile Enhanced and lists two antibodies with Enhanced IHC validation (HPA tissue IHC: reliability; HPA antibodies: HPA009026 and HPA011924). These observations support the reference pattern; they do not validate every catalog antibody or every staining run (HPA antibodies: validation status).
IF/ICC Q: Where should RRBP1 appear?A: At the endoplasmic reticulum (HPA subcellular ICC-IF: enhanced ER location). This is a localisation cross-check for interpreting IHC; the ICC-IF record supplies no IHC protocol conditions (HPA subcellular ICC-IF: location and scope).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
The reported high-staining cells are present, but the section is blank.A staining-run failure is possible; a blank result alone cannot identify which step failed (general IHC practice).Check a concurrently processed positive section, then review primary-antibody use, detection reagents and chromogen development (general IHC practice). Use an HPA-reported high-staining cell population as the biological reference (HPA tissue IHC: positive cell list).
Most apparent signal is nuclear.The dominant compartment disagrees with ER membrane localisation and HPA's cytoplasmic tissue profile (UniProt Q9P2E9: location; HPA tissue IHC: profile).Recheck nuclear counterstain versus chromogen, then compare with a detection control and a reported positive cell population before scoring the result (general IHC practice; HPA tissue IHC: positive cell list).
Unexpected cells stain as strongly as, or more strongly than, the reported positive cells.Cross-reactivity or endogenous detection activity are possibilities; the slide pattern does not prove either cause (general IHC practice).Verify cell identity and compare a suitable detection control. Interpret intensity by cell population using the reported high and low observations (general IHC practice; HPA tissue IHC: positive and low lists).
Color is diffuse or appears outside cells.Nonspecific background or excess chromogen development can obscure cellular localisation (general IHC practice).Inspect controls, blocking and wash steps, then assess development time in a repeat run (general IHC practice). Require a recognizable cytoplasmic cellular pattern for interpretation (HPA tissue IHC: cytoplasmic profile).
Cardiomyocytes, skeletal myocytes or breast adipocytes show faint staining.Low staining is reported for these cell populations (HPA tissue IHC: low list). Faint signal alone does not show that the assay failed.Compare them with an HPA-reported high-staining population in a suitably processed section; record intensity for each identified cell type (HPA tissue IHC: positive and low lists; general IHC practice).
IHC looks broadly cytoplasmic, while an IF/ICC image shows a clearer ER pattern.HPA reports cytoplasmic tissue IHC and enhanced ER localisation by ICC-IF (HPA tissue IHC: profile; HPA subcellular ICC-IF: main location). The assays show localisation at different visual resolution (general IHC/IF practice).Score IHC by its cellular cytoplasmic pattern and tissue context; use the ER-localised IF/ICC observation as a compartment check (HPA tissue IHC: profile; HPA subcellular ICC-IF: main location).

Sample controls for RRBP1 IHC & IF

🧪Run bronchus first and assess its respiratory epithelial cells, which HPA scores High for RRBP1 (HPA: Bronchus, respiratory epithelial cells, High). HPA detects RRBP1 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and expect cells without specific staining on the positive slide to show only background without assigning a named cell type as negative (HPA: no negative rows; RRBP1 detected in all 45 tissues).
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: None in HPA: RRBP1 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 RRBP1 in A-431, U-251MG, U2OS, U2OS, siRNA 2 (10x), U2OS, siRNA 2 (40x), U2OS, scrambled (10x), U2OS, scrambled (40x), with annotated localisation: Endoplasmic reticulum (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section; a concentration-matched, nonimmune rabbit IgG isotype section; and RRBP1 knockout material as a biological specificity control (selected-SKU caption: rabbit primary antibody; standard IHC control practice). Quench endogenous peroxidase and block endogenous biotin when using the caption’s avidin–biotin, DAB detection method (selected-SKU caption: biotinylated secondary, SABC and DAB; standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). The documented IHC example used citrate retrieval at pH 6 for 20 minutes, but whether RRBP1 staining requires that retrieval is unreported (selected-SKU caption: citrate retrieval, pH 6, 20 minutes). Relative ease of frozen-section IHC versus IF is unreported; HPA documents RRBP1 ICC-IF localisation to the endoplasmic reticulum in listed cell lines (HPA subcellular: endoplasmic reticulum, ICC-IF images).

HPA tissue IHC evidence for RRBP1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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

TissueCell typeLevelEvidenceSource
Appendix Endocrine cells High Protein (IHC) HPA →
Bone marrow Hematopoietic cells High Protein (IHC) HPA →
Bronchus Respiratory epithelial cells High Protein (IHC) HPA →
Cerebellum Purkinje cells High Protein (IHC) HPA →
Cerebral cortex Glial cells High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
None in HPA: RRBP1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
Section 3

Advanced RRBP1 IHC Tips

Troubleshoot RRBP1 staining in paraffin sections by checking retrieval, cytoplasmic localisation, detection controls and cell type before comparing signal intensity.

What retrieval should I start with for RRBP1 in paraffin sections?
Start with heat mediated citrate retrieval at pH 6 for 20 minutes in RRBP1 paraffin sections (datasheet A07074-1). The selected tissue image used this retrieval before incubation with 1 μg/ml antibody overnight at 4°C (datasheet A07074-1). Compare a known staining section across runs and inspect whether the cytoplasmic pattern remains distinct from background (HPA: cytoplasmic expression; standard IHC practice). If staining remains weak, test an EDTA retrieval condition on an adjacent section while keeping antibody concentration and detection constant (standard IHC practice). Excessive heating can damage morphology, so assess tissue preservation alongside signal (standard IHC practice).
How should I investigate weak RRBP1 staining when fixation varies?
The selected RRBP1 image identifies paraffin embedded human intestinal cancer tissue, but its caption does not state the fixative (datasheet A07074-1). Target specific fixation sensitivity is therefore unknown from the supplied evidence; do not assign weak staining to a particular fixative without comparison. Record the fixative, fixation duration and processing history for each specimen, then stain matched sections in the same run (standard IHC practice). Hold citrate retrieval at pH 6 for 20 minutes while assessing fixation as a variable (datasheet A07074-1). Judge signal together with morphology and a consistently processed comparison section, since processing differences can confound staining comparisons (standard IHC practice).
Where should a convincing RRBP1 chromogenic signal appear?
Expect predominantly cytoplasmic staining in tissue sections, consistent with the reported tissue profile and endoplasmic reticulum localisation (HPA: cytoplasmic expression in all tissues; HPA: enhanced endoplasmic reticulum location). RRBP1 has a transmembrane segment at residues 8–28 and a cytoplasmic region at 29–1410 (UniProt Q9P2E9 topology). A perinuclear or reticular cytoplasmic pattern is therefore plausible, although chromogenic sections may not resolve individual endoplasmic reticulum structures (UniProt Q9P2E9 localisation; standard IHC practice). Compare cellular staining with tissue morphology and include a section stained without primary antibody to assess detection background (standard IHC practice). Isolated nuclear colour warrants scrutiny because it does not match the supplied localisation evidence (HPA: endoplasmic reticulum; UniProt Q9P2E9 topology).
Could epitope placement or RRBP1 isoforms explain inconsistent staining?
RRBP1 has 2 annotated isoforms, while the supplied antibody caption does not identify the recognised epitope (UniProt Q9P2E9 isoforms; datasheet A07074-1). Its short lumenal segment spans residues 1–7, and the region after the transmembrane segment is cytoplasmic (UniProt Q9P2E9 topology). Without an epitope map, staining cannot establish which isoform or membrane facing region the antibody detects (UniProt Q9P2E9 isoforms and topology). Check the antibody's documented immunogen or epitope before attributing differences to splicing or phosphorylation; RRBP1 has annotated phosphosites, including serines 159 and 165 (UniProt Q9P2E9 modified residues). Compare independent antibody evidence or a validated loss of target control when specificity remains uncertain (standard IHC practice).
How can IF help assess an ambiguous RRBP1 IHC pattern?
For the separate IF/ICC workflow, compare RRBP1 with an endoplasmic reticulum marker and a marker for the cell population being examined (HPA: enhanced endoplasmic reticulum location; standard IF practice). Choose spectrally separated fluorophores and favour a far red channel when tissue autofluorescence obscures shorter wavelengths; inspect unstained and single stain controls (standard IF practice). RRBP1's region after residue 28 faces the cytoplasm, so a confirmed epitope there calls for permeabilisation that permits intracellular antibody access (UniProt Q9P2E9 topology; standard IF practice). Because the antibody epitope is unspecified, optimise permeabilisation against morphology and signal rather than assuming its membrane facing side (datasheet A07074-1; standard IF practice). HPA reports RRBP1 IF localisation to the endoplasmic reticulum (HPA: subcellular location).
How do I distinguish RRBP1 signal from chromogenic background?
The selected image used 10% goat serum blocking, a biotinylated secondary antibody, streptavidin biotin complex and DAB (datasheet A07074-1). Run a no primary control to reveal signal from secondary reagents, endogenous biotin or endogenous peroxidase (standard IHC practice). If diffuse colour persists, check the peroxidase block and consider an avidin biotin block for this detection scheme (standard IHC practice). Compare background before increasing antibody concentration; the demonstrated incubation was 1 μg/ml overnight at 4°C (datasheet A07074-1). Evaluate whether remaining colour follows cytoplasmic cell outlines rather than tissue folds, precipitate or section edges (HPA: cytoplasmic expression; standard IHC practice).
How should I score RRBP1 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable tissue area before scoring, because RRBP1 staining is reported across tissues but varies among cell types (HPA: tissue IHC profile). For cellular intensity, record the percentage of cells at each score from 0 to 3 and calculate an H-score from 0 to 300 (standard IHC practice). Alternatively, report percentage positive cells or positive cell density per mm² of viable tissue, with the threshold stated in advance (standard IHC practice). Normalise to the relevant counted cells or viable area, and exclude necrosis and folds consistently (standard IHC practice). Use the same retrieval, detection and scoring rules across sections being compared (standard IHC practice).
What makes an apparent RRBP1 positive cell credible?
A credible signal is cellular and cytoplasmic, consistent with RRBP1's endoplasmic reticulum location and the reported tissue IHC profile (HPA: enhanced endoplasmic reticulum location; HPA: cytoplasmic expression). Check cell identity against morphology: high staining is reported in colon endocrine cells, whereas breast adipocytes are listed as low (HPA: High in colon endocrine cells; HPA: Low in breast adipocytes). Treat colour concentrated at section edges, folds or necrotic areas as suspect until a clean adjacent area supports it (standard IHC practice). Nuclear only signal conflicts with the supplied localisation evidence, and colour in a no primary control points to a detection artefact (HPA: endoplasmic reticulum; standard IHC practice). Confirm uncertain calls with a consistently stained comparison section (standard IHC practice).
Boster reagents

Best RRBP1 / Ribosome-binding protein 1 IHC Antibodies

The catalog lists two anti-RRBP1 antibodies with IHC images from human paraffin sections and IF images from cultured cells; one lists human, mouse and rat reactivity, and the other human only (catalog images and reactivity).

Real IHC data IHC analysis of RRBP1 using anti-RRBP1 antibody (A07074-1). RRBP1 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-RRBP1 Antibody (A07074-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Anti-RRBP1 Antibody ®
Cat # A07074-1
Real IHC data IHC analysis of RRBP1 using anti-RRBP1 antibody (A07074-2). RRBP1 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-RRBP1 Antibody (A07074-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-RRBP1 Antibody ®
Cat # A07074-2

A07074-1 has IHC images from human intestinal and lung cancer paraffin sections, plus an IF/ICC cell image (A07074-1 image captions). A07074-2 has IHC images from human liver and lung cancer paraffin sections, plus an IF/ICC image of U2OS cells (A07074-2 image captions).

Which to pick: For paraffin-section IHC, choose A07074-1 if its citrate pH 6 retrieval example fits your workflow, or A07074-2 for its EDTA pH 8.0 example; neither caption reports the fixative (A07074-1 and A07074-2 IHC image captions). For human IF/ICC, both list those applications and show cell images; A07074-2 specifically shows U2OS cells (catalog applications; A07074-1 and A07074-2 IF image captions). For mouse or rat samples, choose A07074-1 because it lists those species, while A07074-2 lists human only; clonality is unreported for both (catalog reactivity and clone fields).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry Q9P2E9 (RRBP1_HUMAN, Ribosome-binding protein 1).
  2. Human Protein Atlas. RRBP1 tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. RRBP1 subcellular location (ICC-IF): Localized to the endoplasmic reticulum..
  4. Human Protein Atlas. RRBP1 antibody validation summary (2 antibodies).
  5. Expression of RRBP1 in epithelial ovarian cancer and its clinical significance. Bioscience reports 2019 — PMC6646231.
  6. RRBP1 overexpression is associated with progression and prognosis in endometrial endometrioid adenocarcinoma. Diagnostic pathology 2019 — PMC6347782.
  7. Overexpression of ribosome binding protein 1 (RRBP1) in breast cancer. Clinical proteomics 2012 — PMC3439379.
  8. ALK oncoproteins in atypical inflammatory myofibroblastic tumours: novel RRBP1-ALK fusions in epithelioid inflammatory myofibroblastic sarcoma. The Journal of pathology 2017 — PMC5337428.
  9. PubMed PMID:9628588 — UniProt-cited evidence.
  10. PubMed PMID:10718198 — UniProt-cited evidence.
  11. PubMed PMID:12168954 — UniProt-cited evidence.