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RAB11FIP3 / Rab11 family-interacting protein 3 · IHC design guide
Design Immunohistochemistry for RAB11FIP3
Plan RAB11FIP3 paraffin-section IHC around cytoplasmic staining in kidney tubular cells (HPA tissue IHC). This guide covers consistent fixation, the catalog antibody’s 1:100–1:300 IHC dilution range (datasheet), and controls for presumed off-target binding (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RAB11FIP3 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 (HPA tissue IHC); endosomal and Golgi localization (UniProt) | |
| Staining pattern | Tubular and GI glandular cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding may confound staining (HPA tissue IHC) | |
| Regulation | Expression regulation is unannotated (UniProt) | |
| Isoform / epitope | Three isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended RAB11FIP3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with a published IHC method for human papillary thyroid carcinoma sections (PMC6126170).
| Sample | Paraffin-embedded human lung carcinoma tissue; fixative not specified (datasheet A09469-1) |
| Fixation | Image fixative and duration unreported (datasheet A09469-1); 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-RAB11FIP3, 1:100 - 1:300 (datasheet A09469-1) |
| 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 | RAB11FIP3-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues including cells in tubules and glandular cells in the gastrointestinal tract. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval); the published method uses an autoclave (PMC6126170).
Section 2
What Is the Expected RAB11FIP3 Staining Pattern?
RAB11FIP3 is an intracellular protein associated with recycling endosomes, the centrosome, Golgi membranes and structures involved in cell division (UniProt O75154 subcellular location; UniProt O75154 topology: no transmembrane segment). In tissue IHC, expect cytoplasmic staining, most prominently in kidney tubular cells; several glandular and epithelial populations show medium staining (HPA tissue IHC). HPA rates the tissue pattern Enhanced, while noting medium agreement with RNA data and disregarded presumed off-target binding (HPA tissue IHC).
| Kidney tubular cells show clear cytoplasmic chromogen, with weaker or absent staining in nearby cells. | This matches the strongest listed tissue result: High in kidney tubular cells (HPA tissue IHC). Compare the stain within identifiable tubules rather than treating every cell on the section as an equally strong positive; HPA reports staining by cell population (HPA tissue IHC). |
| Glandular or epithelial cells show moderate cytoplasmic staining outside the kidney. | Medium staining is reported in adrenal, colon, duodenum, fallopian tube and gallbladder glandular cells, bronchial respiratory epithelium and hepatocytes (HPA tissue IHC). A moderate result in these populations can fit the reported pattern; intensity alone does not establish antibody specificity (HPA tissue IHC: presumed off-target binding disregarded). |
| Strong, predominantly nuclear staining appears, with little cytoplasmic signal. | A nuclear-dominant result conflicts with HPA’s cytoplasmic tissue profile and UniProt’s intracellular membrane and cytoskeletal locations (HPA tissue IHC; UniProt O75154 subcellular location). Treat it as suspect and check the control section and detection background before assigning it to RAB11FIP3 (general IHC practice). |
| Chromogen appears in a cell population reported as undetected, or broadly across unlike cell types. | For example, HPA reports no detection in adipocytes or esophageal squamous epithelial cells (HPA tissue IHC). Unexpected staining warrants a cross-reactivity or endogenous detection-activity check; an HPA ‘Not detected’ result is a comparison point, not proof that every specimen of that cell type must be negative (HPA tissue IHC; general IHC practice). |
| A diffuse haze covers cells and extracellular areas, obscuring cell boundaries. | A field-wide haze cannot be scored as the cell-associated cytoplasmic pattern reported by HPA (HPA tissue IHC). Compare a no-primary control, inspect wash and blocking steps, and assess whether excessive chromogen development or counterstain is hiding the distribution (general IHC practice). |
💡Expected RAB11FIP3 appearanceCall the IHC result positive when kidney tubular cells show clear, cell-associated cytoplasmic staining at the reported High level (HPA tissue IHC); strong nuclear-dominant colour or field-wide haze is suspect against that pattern (HPA tissue IHC; general IHC practice).
| Tissue and cell selection | Kidney tubular cells provide the clearest reported positive population, at High staining (HPA tissue IHC). The listed Medium populations offer a less intense comparison, while HPA’s ‘Not detected’ populations can help reveal unexpected colour; these are cell-specific observations, not whole-organ guarantees (HPA tissue IHC). |
| Strength of tissue evidence | The HPA tissue IHC profile is rated Enhanced, but its description reports medium consistency with RNA expression and presumed off-target binding that was disregarded (HPA tissue IHC). Interpret borderline or unexpected staining with controls and cell morphology rather than using the rating as a guarantee for an individual slide (general IHC practice). |
| Intracellular distribution | UniProt places RAB11FIP3 at recycling endosomes, centrosomes, Golgi-associated membranes and, during cytokinesis, the furrow and midbody (UniProt O75154 subcellular location). Routine chromogenic tissue IHC supports a cytoplasmic assessment (HPA tissue IHC); a tiny punctum need not be resolved for the section to be interpretable (general IHC practice). |
| Cell-cycle context | UniProt describes changing distribution across mitosis, with vesicles moving toward the cleavage furrow and midbody late in division (UniProt O75154 subcellular location). A rare dividing cell can therefore differ from a typical interphase cell, but the reported tissue-level expectation remains cytoplasmic staining in the named cell populations (HPA tissue IHC). |
| Isoforms and epitope coverage | Three isoforms are listed (UniProt O75154 isoforms). No antibody epitope or isoform coverage is supplied here, so neither a missing signal nor an unusual distribution can be assigned to isoform recognition from these records alone (UniProt O75154 isoforms; supplied antibody-validation summary). |
| IF/ICC Q&A: what distribution should fluorescence show? | HPA reports mainly vesicles, centriolar satellites and the basal body, with additional cytokinetic-bridge and primary-cilium localisation (HPA subcellular ICC-IF). Those are fluorescence observations; they do not set a required punctate appearance for chromogenic tissue IHC (HPA subcellular ICC-IF; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in the kidney tubular cells selected as the positive control. | The result conflicts with HPA’s High tubular-cell staining, but the source does not identify a target-specific fixation or retrieval failure (HPA tissue IHC). A run-level staining failure remains possible (general IHC practice). | Confirm tubules are present, inspect the control run, and verify primary-antibody, retrieval and detection steps against the assay instructions (general IHC practice). Do not infer that RAB11FIP3 is absent from this negative slide alone. |
| Only weak colour appears in a tissue expected to stain. | The comparison may be a Medium or Low HPA cell population rather than the High kidney tubular population (HPA tissue IHC). Weak colour may also reflect the staining run (general IHC practice). | Identify and score the relevant cell type first, then compare it with a kidney tubular control stained in the same run (HPA tissue IHC; general IHC practice). Review retrieval and detection conditions as general assay parameters, without assuming RAB11FIP3-specific fixation sensitivity. |
| Nuclei dominate the signal. | That distribution conflicts with the reported cytoplasmic tissue pattern and UniProt locations (HPA tissue IHC; UniProt O75154 subcellular location). Background or cross-reactivity is a possible explanation, not a demonstrated diagnosis (general IHC practice). | Check the no-primary control, distinguish chromogen from the nuclear counterstain, and compare cytoplasmic staining in the positive-control tubules (general IHC practice; HPA tissue IHC). Withhold a positive call if the expected cell-associated pattern is missing. |
| Unexpected colour appears in adipocytes or another HPA-undetected population. | Adipocytes are reported as Not detected (HPA tissue IHC). Cross-reactivity or endogenous enzyme activity could contribute to a chromogenic signal (general IHC practice). | Inspect no-primary and appropriate detection controls, review endogenous-activity blocking, and compare the distribution with a same-run positive tissue (general IHC practice). Record the cell type and discrepancy rather than labelling an entire tissue positive. |
| Brown haze appears over tissue and blank areas. | Widespread background can arise from nonspecific reagent binding, inadequate washing or excessive detection development (general IHC practice); it does not reproduce HPA’s cell-associated cytoplasmic profile (HPA tissue IHC). | Compare blank areas and a no-primary section, then review blocking, washes and development time (general IHC practice). Score RAB11FIP3 only where cellular boundaries and the expected compartment remain distinguishable (HPA tissue IHC; general IHC practice). |
| A small dot or bridge-like signal appears in an occasional dividing cell. | Centrosomal and cytokinetic locations are described by UniProt, while bridge localisation is also reported in ICC-IF (UniProt O75154 subcellular location; HPA subcellular ICC-IF). Chromogenic section morphology alone may not identify those structures securely (general IHC practice). | Assess the cell’s morphology and surrounding cytoplasmic pattern, then retain a compartment-specific claim only if the structure is distinguishable (general IHC practice). Use the HPA tissue cell populations to judge the IHC result overall (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing RAB11FIP3 staining
A09469-1 · IHCImmunohistochemistry analysis of paraffin-embedded human lung carcinoma tissue, using RAB11FIP3 Antibody. The picture on the right is blocke
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000090565-RAB11FIP3/tissue/cerebral+cortex#imid_7208688
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000090565-RAB11FIP3/tissue/colon#imid_7208262
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000090565-RAB11FIP3/tissue/liver#imid_7208271
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000090565-RAB11FIP3/tissue/kidney#imid_7208264
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA028631; green = RAB11FIP3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA028631; green = RAB11FIP3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RAB11FIP3 IHC & IF
🧪Run kidney first: cells in tubules should stain (HPA: High in kidney tubule cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the kidney slide, compare tubular staining with background-only cells outside the tubules, without assuming those cells are biologically negative (HPA: High in kidney tubule cells).
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RAB11FIP3 in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Vesicles (supported), Centriolar satellite (approved), Basal body (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and immunoglobulin class (standard IHC practice); use a RAB11FIP3 knockout specimen or peptide competition as a biological specificity control (selected SKU caption: peptide-blocked tissue image). Quench endogenous peroxidase for chromogenic IHC and check for endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected SKU’s paraffin-section caption does not state a fixative (selected SKU caption: fixative unreported). Antigen-retrieval dependency and whether frozen sections are easier are unreported; establish retrieval conditions empirically for the IHC-validated antibody (selected SKU caption: paraffin-section IHC; standard IHC practice). IF/ICC has supporting cell images showing mainly vesicles, centriolar satellites and basal body (HPA: subcellular ICC-IF), while kidney-specific staining artefacts are unreported in the supplied evidence.
HPA tissue IHC evidence for RAB11FIP3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RAB11FIP3 IHC Tips
Troubleshoot chromogenic IHC for RAB11FIP3 in paraffin sections using the catalog image, tissue staining patterns, and known intracellular localisation (catalog caption A09469-1; HPA tissue IHC; UniProt O75154).
How should I adjust retrieval if RAB11FIP3 staining is weak?
Start with citrate buffer at pH 6.0 for 20 min at 95–98 °C (page retrieval rule: cytoplasmic / membrane antigen). Compare that condition with a no-retrieval section from the same block, keeping antibody dilution and chromogen development constant so the effect of retrieval is interpretable (standard IHC practice). Score cytoplasmic signal in kidney tubule cells alongside background in adjacent tissue; kidney tubule cells show high staining in the reference profile (HPA tissue IHC: High in kidney tubule cells). If staining remains weak, test EDTA at pH 9.0 as a fallback on a separate section, and reject conditions that increase diffuse background or damage tissue morphology (standard IHC practice).
Could fixation explain a weak or uneven RAB11FIP3 result?
RAB11FIP3-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section image does not report a fixative (catalog caption A09469-1). Record the fixative, fixation duration, processing history, and block age for each specimen before comparing staining across cases (standard IHC practice). If fixation is suspected, stain sections from differently processed blocks in one run with the same pH 6.0, 20 min retrieval condition and identical detection settings (page retrieval rule: cytoplasmic / membrane antigen; standard IHC practice). Interpret a between-block difference cautiously, because processing can affect antigen detection and morphology without establishing a RAB11FIP3-specific fixation effect (standard IHC practice).
Where should I expect convincing RAB11FIP3 staining in paraffin sections?
Look primarily for cytoplasmic staining in the relevant cells, with a potentially punctate or perinuclear pattern consistent with recycling endosomes (HPA tissue IHC: cytoplasmic expression; UniProt O75154 subcellular localisation). RAB11FIP3 also localises to centrosomes, the cleavage furrow, and the midbody at particular cell-cycle stages, so these patterns require suitable cellular context rather than routine tissue-wide scoring (UniProt O75154 subcellular localisation). Use kidney tubule cells as a reference population because their reported staining is high, and compare them with the cell population under study on the same run (HPA tissue IHC: High in kidney tubule cells; standard IHC practice). Treat predominantly nuclear or uniformly extracellular chromogen as a reason to recheck antibody specificity, tissue preservation, and detection controls (UniProt O75154 subcellular localisation; standard IHC practice).
How do isoforms and epitope location affect interpretation of staining?
RAB11FIP3 has 3 annotated isoforms, but the supplied catalog caption does not identify the antibody epitope or establish which isoforms it detects (UniProt O75154 isoforms; catalog caption A09469-1). Map the immunogen against each isoform before calling a negative section isoform-negative, and check whether the epitope lies near the EF-hand regions at residues 202–269 or the FIP-RBD at 694–756 (UniProt O75154 domains; standard IHC practice). The protein also has annotated phosphorylation sites, including serine 102, so document epitope overlap when sequence information becomes available (UniProt O75154 modified residues; standard IHC practice). Use an independent epitope or an orthogonal expression assay to investigate discordant staining; the peptide-blocked catalog image alone does not resolve isoform coverage (catalog caption A09469-1; standard IHC practice).
How can IF help resolve an ambiguous chromogenic IHC pattern?
On a separately optimised IF section, multiplex RAB11FIP3 with a validated kidney tubule cell marker when checking the high-staining reference population (HPA tissue IHC: High in kidney tubule cells; standard IF practice). Choose a far-red fluorophore and include an unstained tissue section to assess autofluorescence before interpreting puncta, particularly where endogenous tissue fluorescence is strong (standard IF practice). Because RAB11FIP3 lacks a transmembrane segment and its expected sites include intracellular vesicles, check epitope accessibility and optimise detergent permeabilisation, for example 0.1% Triton X-100 for 5 min (UniProt O75154 topology and subcellular localisation; standard IF practice). Compare the IF distribution with the chromogenic result without assuming that an IF pattern validates paraffin-section IHC specificity (standard IHC / IF practice).
What should I check when chromogen obscures RAB11FIP3 staining?
Inspect a no-primary control and a matched section with the primary antibody to separate detection-system background from candidate RAB11FIP3 signal (standard IHC practice). For peroxidase detection, include a peroxide block, then optimise protein blocking, primary-antibody concentration, wash stringency, and DAB development time one variable at a time (standard chromogenic IHC practice). The selected paraffin-section image includes a peptide-blocked comparison, but its caption gives no fixative or dilution, so those settings must be established locally (catalog caption A09469-1). Preserve interpretable cytoplasmic detail in kidney tubule cells while reducing diffuse stain, since that population has high reported staining and the protein is intracellular (HPA tissue IHC: High in kidney tubule cells; UniProt O75154 subcellular localisation).
How should I score RAB11FIP3 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring rule before reading cases, because the reference tissue profile varies by cell type (HPA tissue IHC: cytoplasmic expression and cell-specific levels; standard IHC practice). For a semiquantitative result, record the percentage of cells at intensities 0–3 and calculate an H-score from 0–300; retain the percentages so intensity and prevalence remain visible (standard IHC practice). For spatial analysis, report positive-cell density per mm² of viable, annotated tissue rather than total section area, excluding folds and necrosis (standard IHC practice). Normalise comparisons to the same eligible cell type, tissue area, batch controls, and fixed chromogen-development settings; report cytoplasmic and focal punctate patterns separately if both are scored (standard IHC practice; UniProt O75154 subcellular localisation).
How can I distinguish true RAB11FIP3 staining from artefact?
Give greatest weight to reproducible cytoplasmic staining in an expected cell population, such as kidney tubule cells, with appropriate negative and detection controls (HPA tissue IHC: High in kidney tubule cells; standard IHC practice). Interpret strong nuclear, extracellular, or indiscriminate tissue-wide staining cautiously because the annotated sites are intracellular vesicles, centrosomes, and related structures (UniProt O75154 subcellular localisation). Check section edges, folds, and necrotic areas for staining that tracks tissue damage rather than intact cells, and check the no-primary control for endogenous enzyme or detection-system signal (standard IHC practice). The peptide-blocked catalog image supports examination of that antibody's staining, while the HPA profile explicitly notes presumed off-target binding; neither observation alone establishes specificity in a new specimen (catalog caption A09469-1; HPA tissue IHC: reliability description).
Boster reagents
Best RAB11FIP3 / Rab11 family-interacting protein 3 IHC Antibodies
A09469-1 is listed for human, mouse and rat IHC, IF and ICC (catalog: applications/reactivity); its pictured evidence shows paraffin-embedded human lung carcinoma IHC (catalog: IHC image caption).
Real IHC data
Anti-RAB11FIP3 Antibody
Cat # A09469-1
A09469-1 is a rabbit polyclonal antibody listed for IHC, IF and ICC in human, mouse and rat (catalog: host/clonality, applications/reactivity). Its own IHC image shows paraffin-embedded human lung carcinoma with synthesized-peptide blocking; no IF image is supplied (catalog: IHC/IF image captions).
Which to pick: Choose A09469-1 for paraffin-section tissue IHC: its own image documents human lung carcinoma, and the listed dilution is 1:100–1:300 (catalog: IHC image caption; datasheet: IHC dilution). For IF/ICC or mouse and rat samples, A09469-1 is listed for those applications and species, with IF at 1:50; its pictured evidence remains human tissue IHC (catalog: applications/reactivity, IF dilution, IHC/IF image captions). The IHC caption specifies paraffin embedding but does not report the fixative (catalog: IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75154 (RFIP3_HUMAN, Rab11 family-interacting protein 3).
- Human Protein Atlas. RAB11FIP3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RAB11FIP3 subcellular location (ICC-IF): Mainly localized to vesicles, centriolar satellites and basal body. In addition localized to the cytokinetic bridge and primary cilium..
- Human Protein Atlas. RAB11FIP3 antibody validation summary (3 antibodies).
- Eps15 homology domain 1 promotes the evolution of papillary thyroid cancer by regulating endocytotic recycling of epidermal growth factor receptor. Oncology letters 2018 — PMC6126170.
- Breast Cancer-Derived Extracellular Vesicles Modulate the Cytoplasmic and Cytoskeletal Dynamics of Blood-Brain Barrier Endothelial Cells. Journal of extracellular vesicles 2025 — PMC11770372.
- PubMed PMID:11481332 — UniProt-cited evidence.
- PubMed PMID:9734811 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RAB11FIP3 (Rab11 family-interacting protein 3). Expected localisation Cytoplasmic staining (HPA tissue IHC); endosomal and Golgi localization (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A09469-1 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung carcinoma tissue; fixative not specified (datasheet A09469-1); 2. Fixation: Image fixative and duration unreported (datasheet A09469-1); 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-RAB11FIP3, 1:100 - 1:300 (datasheet A09469-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target binding may confound staining (HPA tissue IHC); Isoform / epitope: Three isoforms; epitope coverage is unspecified (UniProt); Regulation: Expression regulation is unannotated (UniProt). Evidence: UniProt O75154, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to vesicles, centriolar satellites and basal body.…, Datasheet A09469-1, PMC6126170. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RAB11FIP3/immunohistochemistry-ihc-guide.