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RBP1 / Retinol-binding protein 1 · IHC design guide
Design Immunohistochemistry for RBP1
Plan chromogenic RBP1 IHC in paraffin sections with the IHC-validated antibody A03820-4 (datasheet). Assess cytoplasmic staining in Leydig and ovarian stromal cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RBP1 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 staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in Leydig and ovarian stromal cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03820-4) | |
| Positive control | Testis+4 more · see all | |
| Negative control | Adipose tissue+4 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 A03820-4) | |
| Caveat | Antibody staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Higher expression in adult ovary and pancreas (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended RBP1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published chromogenic tissue protocols for RBP1 (PMC7316825; PMC12481618; PMC10557880).
| Sample | Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A03820-4) |
| Fixation | Image fixative and duration unreported (datasheet A03820-4); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A03820-4); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03820-4) |
| Primary antibody | Rabbit anti-RBP1, 2-5 μg/ml (datasheet A03820-4) |
| Primary incubation | Overnight at 4 °C (datasheet A03820-4) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03820-4) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RBP1-positive staining in leydig cells of testis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, most abundant in Leydig cells and ovarian stromal cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet: A03820-4); the oral cancer protocol used citrate at pH 6.0 (PMC7316825).
Section 2
What Is the Expected RBP1 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic RBP1 staining, strongest in testicular Leydig cells and present at medium levels in ovarian stromal cells, adrenal glandular cells, endometrial stromal cells, fallopian tube ciliated cell bodies, and pancreatic exocrine glandular cells (HPA tissue IHC). RBP1 is a cytoplasmic protein also associated with lipid droplets and has no transmembrane segment (UniProt P09455). HPA rates the tissue profile Enhanced while reporting low agreement between staining and RNA expression (HPA tissue IHC).
| Strong cytoplasmic color in Leydig cells, with lighter staining in the listed medium-level cell populations. | This matches the cell-specific IHC profile: Leydig cells are High; ovarian stromal and the other listed positive cells are Medium (HPA tissue IHC). Compare named cell populations rather than treating every cell in a positive tissue as positive (HPA tissue IHC). |
| Predominantly nuclear staining in a paraffin section, with little cytoplasmic signal. | This does not match the predominantly cytoplasmic tissue IHC profile (HPA tissue IHC). Review staining and counterstain controls before assigning RBP1; ICC-IF separately records an additional nucleoplasmic location, so some nuclear IF signal is plausible (HPA subcellular). |
| Strong color in adipocytes or appendix glandular cells while expected positive cells stain weakly. | Those named populations are Not detected in HPA tissue IHC (HPA tissue IHC). Check antibody specificity and endogenous detection activity with controls; staining in an unexpected cell type alone does not establish which cause applies (general IHC practice). |
| Diffuse color across tissue compartments, including cells expected to be unstained. | A uniform haze obscures the cell-specific RBP1 pattern (HPA tissue IHC). Compare a no-primary control and review blocking, washing, antibody concentration, and chromogen development as general IHC troubleshooting steps (general IHC practice). |
| No cytoplasmic signal in testicular Leydig cells. | Leydig cells are a High-staining reference population (HPA tissue IHC). First confirm tissue identification and that the detection run worked; then review the antibody's stated IHC-P conditions and general retrieval and staining controls (general IHC practice). |
💡Expected RBP1 appearanceCall a result consistent with RBP1 when cytoplasmic staining is High in Leydig cells or Medium in the specified stromal and glandular populations; widespread staining of HPA Not detected cell populations is a specificity warning (HPA tissue IHC).
| Cell population and tissue | Intensity is cell specific: Leydig cells are High, several stromal or glandular populations are Medium, and adipocytes are Not detected (HPA tissue IHC). Score the relevant cells, not the section as a whole (general IHC practice). |
| Antibody validation and RNA agreement | The tissue profile is Enhanced because paired antibodies show a similar protein pattern, yet agreement with RNA expression is low (HPA tissue IHC). Use the observed protein pattern as the IHC comparator without equating RNA abundance with stain intensity. |
| Protein topology and processing | RBP1 has no transmembrane segment, signal peptide, or propeptide, and its annotated chain spans residues 1–135 (UniProt P09455). These annotations support an intracellular pattern; they do not establish an antigen retrieval requirement. |
| IF/ICC Q: Is a nuclear signal expected? | A: HPA records supported cytosolic localisation and an approved additional nucleoplasmic location in ICC-IF (HPA subcellular). Interpret that observation within IF/ICC; the tissue IHC profile remains predominantly cytoplasmic (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Leydig cells are blank. | A High-staining reference population is absent (HPA tissue IHC); the cause cannot be determined from the slide alone. | Verify the testis cell identification, positive control, antibody coverage, and detection run; review the catalog antibody's IHC-P conditions and retrieval steps (general IHC practice). |
| Nuclei dominate the chromogenic signal. | The distribution conflicts with the cytoplasmic tissue IHC profile (HPA tissue IHC); staining or counterstain artefact is possible (general IHC practice). | Inspect the no-primary control and counterstain, then compare cytoplasmic signal in a known positive cell population (general IHC practice). |
| Adipocytes stain strongly. | HPA reports adipocytes as Not detected (HPA tissue IHC); antibody cross-reactivity or endogenous detection activity are possibilities (general IHC practice). | Check a no-primary control and compare the result with Leydig-cell staining; investigate specificity if the unexpected signal persists (general IHC practice). |
| The whole section has diffuse color. | Background can obscure the expected cell-specific pattern (HPA tissue IHC); nonspecific binding or detection background is possible (general IHC practice). | Compare no-primary controls and review blocking, wash steps, antibody concentration, and chromogen development (general IHC practice). |
| Ovarian stromal cells look weaker than Leydig cells. | This may match the reference: ovarian stromal cells are Medium and Leydig cells are High (HPA tissue IHC). | Score each named population against its own expected level and cytoplasmic location before calling a run weak (HPA tissue IHC; general IHC practice). |
| Protein staining and RNA abundance disagree. | HPA explicitly reports low consistency between antibody staining and RNA expression, despite an Enhanced protein profile (HPA tissue IHC). | Report the discrepancy and assess the IHC result using cell-level staining, controls, and the paired-antibody validation context (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing RBP1 staining
A03820-4 · IHCIHC analysis of RBP1 using anti-RBP1 antibody (A03820-4). RBP1 was detected in a paraffin-embedded section of human glioblastoma tissue. Hea
A03820-4 · IHCIHC analysis of RBP1 using anti-RBP1 antibody (A03820-4). RBP1 was detected in a paraffin-embedded section of human liver cancer tissue. Hea
A03820-4 · IFIF analysis of RBP1 using anti-RBP1 antibody (A03820-4) and anti-Tubulin Alpha antibody (M03989-3). RBP1 was detected in immunocytochemical
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000114115-RBP1/tissue/cerebral+cortex#imid_17814291
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000114115-RBP1/tissue/colon#imid_17814217
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000114115-RBP1/tissue/liver#imid_17814202
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000114115-RBP1/tissue/kidney#imid_17814211
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA007338; green = RBP1, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA007338; green = RBP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RBP1 IHC & IF
🧪Run testis first and require staining in Leydig cells (HPA: High in testis Leydig cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); neighboring testis cells chosen as internal negatives should show only background chromogen, without specific cytoplasmic staining (standard IHC practice; UniProt P09455: cytoplasm).
Positive control tissue: Testis (Leydig cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RBP1 in Hep-G2, SH-SY5Y, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary secondary-only slide, a rabbit IgG control matched to the primary antibody’s clonality and isotype, and an RBP1 knockout or immunizing-peptide competition control if available (A03820-4 tissue-IHC caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase in testis before HRP/DAB detection and compare background with the no-primary slide (A03820-4 tissue-IHC caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state a fixative (A03820-4 tissue-IHC caption: fixative not stated). That example used heat retrieval in EDTA at pH 8.0, but the evidence does not establish retrieval dependence or show that frozen sections or IF are easier (A03820-4 tissue-IHC caption: heat retrieval in EDTA; HPA subcellular: ICC-IF images). Leydig-cell pigment can resemble brown DAB, so check the no-primary slide when scoring testis (standard IHC practice).
HPA tissue IHC evidence for RBP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Low consistency between antibody staining and RNA expression data. Paired antibodies with high similarity support the protein profile.). 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 |
|---|---|---|---|---|
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RBP1 IHC Tips
Troubleshoot RBP1 staining in paraffin sections by checking retrieval, compartment, cell identity, and controls before comparing signal across samples.
Which retrieval condition should I start with for RBP1 paraffin-section IHC?
Start with heat-mediated retrieval in EDTA at pH 8.0 for the catalog antibody (datasheet A03820-4). The demonstrated paraffin-section workflow then used 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A03820-4). If staining is weak, compare retrieval duration on matched sections while holding detection and exposure to chromogen constant (standard IHC practice). Inspect tissue integrity and background alongside cytoplasmic signal; stronger overall DAB staining alone does not establish improved RBP1 detection (UniProt P09455 subcellular location; standard IHC practice).
How should I troubleshoot fixation when RBP1 staining is weak?
The selected paraffin-section caption does not report a fixative, so target-specific fixation sensitivity is unknown (datasheet A03820-4). Record fixative, fixation interval, and processing history for each specimen, then compare sections processed together using the same retrieval and antibody conditions (standard IHC practice). Include a positive control with the expected cell-specific pattern, such as Leydig cells, when available (HPA: High in Leydig cells). If one processing batch loses cytoplasmic staining while its matched controls perform normally, investigate handling and retrieval before attributing the difference to RBP1 biology (UniProt P09455 subcellular location; standard IHC practice).
Where should convincing RBP1 staining appear in a tissue section?
Prioritize cytoplasmic staining in the expected cell population: RBP1 is cytoplasmic and also associated with lipid droplets (UniProt P09455 subcellular location). Tissue IHC reports cytoplasmic expression, with particularly strong staining in Leydig cells and staining in ovarian stromal cells (HPA tissue IHC profile). Score the cell body separately from nuclei and extracellular material, using morphology and a counterstain to assign signal to cells (standard IHC practice). HPA also reports nucleoplasmic localization in cell imaging, so an isolated nuclear DAB pattern should prompt control and antibody-specificity checks before interpretation (HPA subcellular: approved nucleoplasm; standard IHC practice).
Could isoforms or epitope accessibility explain discordant RBP1 staining?
RBP1 has 3 annotated isoforms, but the supplied evidence does not map this antibody’s epitope to any isoform (UniProt P09455 isoforms; datasheet A03820-4). The annotated protein has 135 residues, no signal peptide or transmembrane segment, and methylation at residue 9 (UniProt P09455 processing, topology, modified residues). Compare the immunogen or epitope documentation with isoform sequences before claiming isoform-specific loss; retrieval can also change epitope accessibility in paraffin sections (standard IHC practice). When staining differs across tissues, confirm the pattern with a second antibody recognizing a distinct documented epitope and appropriate cell-level controls (standard IHC practice).
How should I adapt the RBP1 question for multiplex IF?
For the separate IF/ICC assay, pair RBP1 with a marker identifying the expected cell population, such as Leydig cells in testis, and check signal within individual cells (HPA: High in Leydig cells). Choose fluorophores after examining unstained tissue for autofluorescence, reserving a cleaner channel for the weaker signal (standard IF practice). Because RBP1 is cytoplasmic and has no transmembrane segment, use permeabilisation appropriate for access to an intracellular epitope, then verify that morphology and signal remain intact (UniProt P09455 topology and subcellular location; standard IF practice). HPA reports cytosolic and additional nucleoplasmic IF localization, which can guide compartment checks without establishing this catalog antibody’s IF performance (HPA subcellular; datasheet A03820-4).
What should I change when DAB background obscures RBP1?
First compare a no-primary control with the stained section to distinguish detection-system signal from primary-antibody-dependent staining (standard IHC practice). In the documented workflow, sections received 10% goat serum block, 2 μg/ml primary overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet A03820-4). Check peroxidase blocking, washes, and DAB development on matched sections; these are general chromogenic workflow controls, not evidence of an RBP1-specific effect (standard IHC practice). Judge background against the expected cytoplasmic cell pattern, including HPA-negative adipocytes where relevant, rather than against overall section color (UniProt P09455 subcellular location; HPA: adipocytes Not detected).
How should I quantify RBP1 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; RBP1 tissue staining is reported as cytoplasmic, including High expression in Leydig cells (HPA tissue IHC). For chromogenic sections, record percentage of positive target cells and staining intensity, then calculate an H-score using consistent thresholds (standard IHC practice). Alternatively, measure positive-cell density per mm² when cell counts per area answer the study question, excluding necrosis and folds by a documented rule (standard IHC practice). Normalize each result to the number or area of eligible target cells, and compare batches against the same reference control because staining conditions affect measured intensity (standard IHC practice).
How can I distinguish true RBP1 signal from artefact?
A plausible positive follows cellular morphology and is predominantly cytoplasmic in a cell population supported by tissue evidence, such as Leydig cells (UniProt P09455 subcellular location; HPA: High in Leydig cells). Treat strong staining confined to tissue edges, necrotic regions, or extracellular deposits as suspect until it persists in well-preserved central tissue (standard IHC practice). Investigate isolated nuclear signal carefully: nucleoplasmic localization is reported in cell imaging, but tissue IHC is described chiefly as cytoplasmic (HPA subcellular; HPA tissue IHC profile). Compare no-primary and peroxidase-block controls to detect endogenous enzyme or detection background, and weigh discrepant results against HPA’s reported low staining–RNA consistency (standard IHC practice; HPA reliability description).
Boster reagents
Best RBP1 / Retinol-binding protein 1 IHC Antibodies
The catalog antibody has IHC images from human and mouse paraffin sections and an IF image from U2OS cells (IHC and IF captions); rat reactivity is listed without an image (catalog: reactivity).
Real IHC data
Anti-RBP1 Antibody ®
Cat # A03820-4
A03820-4 has IHC images from paraffin sections of human glioblastoma, liver cancer and rectum adenocarcinoma, and mouse ovary (IHC captions). A03820-4 also has an IF/ICC image from U2OS cells (IF caption), with human, mouse and rat reactivity listed (catalog: reactivity).
Which to pick: For tissue IHC, choose A03820-4: its paraffin-section captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (IHC captions). For IF/ICC, choose A03820-4 based on its U2OS image and listed IF/ICC applications (IF caption; catalog: applications). For work across species, A03820-4 lists human, mouse and rat reactivity, while its IHC images cover human and mouse; clonality is unreported.
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P09455 (RET1_HUMAN, Retinol-binding protein 1).
- Human Protein Atlas. RBP1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RBP1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm..
- Human Protein Atlas. RBP1 antibody validation summary (3 antibodies).
- The RBP1-CKAP4 axis activates oncogenic autophagy and promotes cancer progression in oral squamous cell carcinoma. Cell death & disease 2020 — PMC7316825.
- CircSPINT2 confers sensitivity to osimertinib via hsa-miR-1296-3p/RBP1 axis and inhibits NSCLC progression. Molecular therapy. Oncology 2025 — PMC12481618.
- Hepatic stellate cell hypertrophy is associated with metabolic liver fibrosis. Scientific reports 2020 — PMC7052210.
- ELOA promotes tumor growth and metastasis by activating RBP1 in gastric cancer. Cancer medicine 2023 — PMC10557880.
- PubMed PMID:2992469 — UniProt-cited evidence.
- PubMed PMID:3356192 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RBP1 (Retinol-binding protein 1). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03820-4). Validated IHC antibody: A03820-4 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Leydig cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A03820-4); 2. Fixation: Image fixative and duration unreported (datasheet A03820-4); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A03820-4); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03820-4); 8. Primary antibody: Rabbit anti-RBP1, 2-5 μg/ml (datasheet A03820-4); 9. Primary incubation: Overnight at 4 °C (datasheet A03820-4). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03820-4); Caveat: Antibody staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt); Regulation: Higher expression in adult ovary and pancreas (UniProt). Evidence: UniProt P09455, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A03820-4, PMC7316825, PMC12481618. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RBP1/immunohistochemistry-ihc-guide.