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RAB11A / Ras-related protein Rab-11A · IHC design guide
Design Immunohistochemistry for RAB11A
Plan chromogenic RAB11A IHC-P around cytoplasmic and membranous tissue staining (HPA tissue IHC). This guide covers fixation, epithelial positive controls and cell populations that may show no signal (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RAB11A 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 and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular and squamous cells: cytoplasmic and membranous (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M01436-2) | |
| Positive control | Adrenal gland+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 consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Some cell populations, including adipocytes, show no signal (HPA tissue IHC) | |
| Regulation | No expression regulation annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage unknown; no transmembrane segment (UniProt) |
Section 1
Recommended RAB11A IHC & IF Protocols
The catalog antibody’s IHC protocol (datasheet: M01436-2) is followed by published RAB11A staining protocols for lung and colon tissues (PMC12376738; PMC6726494; PMC5438609; PMC6605470).
| Sample | Paraffin-embedded human spleen tissue; fixative not specified (datasheet M01436-2) |
| Fixation | Image fixative and duration unreported (datasheet M01436-2); 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 M01436-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M01436-2) |
| Primary antibody | Mouse monoclonal (clone 4H9) anti-RAB11A, 2 μg/ml (datasheet M01436-2) |
| Primary incubation | Overnight at 4 °C (datasheet M01436-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet M01436-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RAB11A-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Ubiquitous cytoplasmic and membranous expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA, pH 8.0 (datasheet: M01436-2); citrate retrieval is a published alternative for the lung tissue protocols (PMC12376738; PMC5438609).
Section 2
What Is the Expected RAB11A Staining Pattern?
RAB11A should show cytoplasmic and membranous staining, consistent with its association with endosomes, recycling vesicles and the Golgi; it has no transmembrane segment (HPA: ubiquitous cytoplasmic and membranous expression; UniProt P62491: subcellular location and topology). Glandular cells in colon, duodenum and appendix are documented medium-intensity IHC examples (HPA: tissue IHC). HPA rates the tissue pattern Approved, with medium consistency between staining and RNA expression (HPA: reliability).
| Cytoplasmic staining with membranous accentuation in glandular cells of colon or duodenum. | This fits the documented medium-intensity tissue pattern and RAB11A’s membrane-trafficking locations (HPA: tissue IHC; UniProt P62491: subcellular location). Assess the relevant cells and compartments together; HPA’s ubiquitous profile does not imply equal intensity in every cell (HPA: tissue IHC). |
| A sharply nuclear-only signal, with little cytoplasmic or membranous staining. | This conflicts with the reported IHC profile and annotated locations (HPA: tissue IHC; UniProt P62491: subcellular location). Treat it as a suspect pattern and check the antibody and detection controls before interpreting it as RAB11A (general IHC practice). |
| Prominent staining in adipocytes or pancreatic exocrine glandular cells. | HPA reports RAB11A as not detected in those specific cell populations (HPA: tissue IHC). Recheck cell identity and compare controls; cross-reactivity or endogenous detection activity are possible technical explanations (general IHC practice). “Not detected” is an observation, not proof of biological absence. |
| A uniform haze across tissue, stroma and slide background. | The haze does not resolve the reported cytoplasmic and membranous distribution (HPA: tissue IHC). Nonspecific antibody binding, insufficient washing or detection background may obscure a real signal; compare a reagent control on the same run (general IHC practice). |
| No discernible signal in colon glandular cells while the section remains interpretable. | Colon glandular cells are documented at medium staining intensity (HPA: tissue IHC). A blank result warrants a check of tissue quality, antibody conditions and detection controls before concluding that the specimen lacks RAB11A (general IHC practice). |
💡Expected RAB11A appearanceCall a result positive when glandular cells show discernible, approximately medium cytoplasmic and membranous staining, as documented for colon and duodenum (HPA: tissue IHC); isolated nuclear-only colour or uniform background is a suspect result (HPA: tissue IHC; UniProt P62491: subcellular location; general IHC practice).
| Cell and compartment selection | Score the cells identified in the tissue record, not the section as a whole: HPA reports medium staining in colon glandular cells but no detection in pancreatic exocrine glandular cells (HPA: tissue IHC). |
| Antibody validation | The HPA tissue profile is Approved, with medium staining–RNA consistency; HPA060686 and CAB013097 are individually Approved for IHC (HPA: reliability and antibody validation). These ratings support the reported pattern without guaranteeing every specimen will match. |
| Topology and processing | RAB11A has no transmembrane segment and is annotated on several intracellular membranes; its reported processing includes a 214–216 propeptide (UniProt P62491: topology, subcellular location and processing). The supplied record gives no antibody epitope, so it cannot predict epitope access or retrieval needs. |
| IF/ICC Q&A: What pattern should IF show? | Mainly vesicles and centriolar satellites are supported IF locations; primary cilium, basal body and cytosol are marked uncertain (HPA: subcellular ICC-IF). Use the separate IF/ICC guide for its procedure; these IF observations do not set an IHC protocol. |
| Detection background | Endogenous enzyme activity or nonspecific reagent binding can produce chromogenic colour unrelated to the target (general IHC practice). Interpret suspicious staining against appropriate detection and antibody controls; HPA’s tissue observations do not identify its cause. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected glandular-cell signal is absent. | A failed staining run, unsuitable antibody conditions or compromised section is possible; HPA records medium staining in colon and duodenum glandular cells (HPA: tissue IHC; general IHC practice). | Check an HPA-documented positive tissue in the same run, then review the catalog antibody’s IHC-P conditions and detection controls (HPA: tissue IHC; general IHC practice). |
| Strong signal is confined to nuclei. | A nuclear-only pattern conflicts with the reported cytoplasmic and membranous IHC distribution and annotated organelle locations (HPA: tissue IHC; UniProt P62491: subcellular location). | Inspect a reagent control and compare the stained section with an HPA-documented positive-cell pattern before assigning RAB11A positivity (general IHC practice; HPA: tissue IHC). |
| Pancreatic exocrine cells or adipocytes stain strongly. | These particular cells are listed as not detected by HPA; cross-reactivity or chromogenic detection background may explain discordant colour (HPA: tissue IHC; general IHC practice). | Confirm the stained cell type, review controls and assess whether the same signal appears when the primary antibody is omitted (general IHC practice). |
| Colour spreads evenly beyond cell boundaries. | Nonspecific binding, excess staining or incomplete washing can obscure the cellular pattern (general IHC practice). | Compare a matched reagent control, review washing and antibody conditions, and score only resolvable cell-associated staining (general IHC practice). |
| An HPA-listed not-detected cell population has no signal. | A negative result there agrees with that specific HPA observation; it does not establish that all cells in the tissue lack RAB11A (HPA: tissue IHC). | Use a documented medium-staining cell population to verify the run before interpreting the negative population (HPA: tissue IHC; general IHC practice). |
| A proposed antigen-retrieval change is claimed to rescue RAB11A specifically. | Neither the supplied HPA tissue data nor UniProt topology and processing establish a RAB11A-specific fixation or retrieval effect (HPA: tissue IHC; UniProt P62491: topology and processing). | Follow the catalog antibody’s validated IHC-P procedure as the starting point; evaluate any retrieval adjustment with matched controls, without presuming a target-specific benefit (general IHC practice). |
Experiment evidence
IHC & IF images showing RAB11A staining
M01436-2 · IHCIHC analysis of Rab11A using anti-Rab11A antibody (M01436-2). Rab11A was detected in a paraffin-embedded section of human spleen tissue. Hea
M01436-2 · IHCIHC analysis of Rab11A using anti-Rab11A antibody (M01436-2). Rab11A was detected in a paraffin-embedded section of human gastric carcinoma
M01436-2 · IFIF analysis of Rab11A using anti-Rab11A antibody (M01436-2). Rab11A was detected in an immunocytochemical section of T-47D cells. Enzyme ant
M01436-1 · IHCM01436-1 staining RAB11A in human brain sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissue
PB9789 · IFIF analysis of Rab11A using anti-Rab11A antibody (PB9789). Rab11A was detected in an immunocytochemical section of U2OS cells. Enzyme antige
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000103769-RAB11A/tissue/cerebral+cortex#imid_17755645
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000103769-RAB11A/tissue/colon#imid_17755577
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000103769-RAB11A/tissue/liver#imid_17755586
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000103769-RAB11A/tissue/kidney#imid_17755579
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB013097; green = RAB11A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB013097; green = RAB11A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RAB11A IHC & IF
🧪Run colon first and expect its glandular cells to stain (HPA: colon glandular cells, Medium). Use smooth muscle as the negative tissue (HPA: smooth muscle cells, Not detected); on a colon section that includes smooth muscle, those cells should show only background signal (HPA: smooth muscle cells, Not detected).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RAB11A in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, HEK293, NIH 3T3, with annotated localisation: Vesicles (supported), Centriolar satellite (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a mouse IgG isotype control matched to the primary antibody (selected-SKU caption: mouse anti-Rab11A), plus a validated RAB11A-knockout biological negative. For the biotin-based DAB method, check endogenous peroxidase and biotin background on the colon section (selected-SKU caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected-SKU caption: EDTA retrieval). IF/ICC can help assess vesicular staining (HPA: supported vesicle localization); the supplied evidence does not establish whether IF/ICC or frozen sections are easier than paraffin IHC, while biotin or peroxidase background should be checked in the proposed colon assay (selected-SKU caption: SABC/DAB detection).
HPA tissue IHC evidence for RAB11A
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | 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 → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RAB11A IHC Tips
Troubleshoot RAB11A staining in paraffin sections by checking retrieval, cell type, and vesicular localisation before comparing signal intensity across samples.
What retrieval should I use when RAB11A staining is weak in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M01436-2). The selected image used that retrieval before staining human spleen with 2 μg/ml antibody overnight at 4°C (caption M01436-2). If signal is weak, first check that sections were heated consistently and that the retrieval solution maintained its intended pH (standard IHC practice). Compare a small retrieval time series while holding antibody concentration and DAB development constant, and assess whether tissue structure remains intact (standard IHC practice). The image caption does not state a fixative, so it cannot establish how a particular fixation condition responds to retrieval (caption M01436-2).
Could fixation explain variable RAB11A staining between paraffin blocks?
Target specific fixation sensitivity is unknown: the selected paraffin section caption does not report its fixative (caption M01436-2). Record each block's fixative and fixation duration, then compare similarly processed sections using the same EDTA pH 8.0 retrieval and detection conditions (datasheet M01436-2; standard IHC practice). Include a consistently processed reference section in each run to distinguish block variation from staining variation (standard IHC practice). If signal differs, check tissue preservation and retrieval consistency before changing antibody concentration (standard IHC practice). Neither the reported tissue staining pattern nor RAB11A's topology establishes a fixation effect for this antibody (HPA: tissue IHC; UniProt P62491 topology).
Which RAB11A staining pattern is plausible in paraffin sections?
Expect mainly cytoplasmic, vesicular and potentially membranous staining, while judging the pattern in the identified cell type (HPA: ubiquitous cytoplasmic and membranous expression; UniProt P62491 subcellular localisation). RAB11A associates with recycling endosomes, other vesicle membranes and the trans Golgi network, and has no transmembrane segment (UniProt P62491 localisation and topology). Perinuclear puncta can fit its endosomal recycling compartment localisation, but a uniformly intense nuclear pattern warrants scrutiny (UniProt P62491 localisation; standard IHC practice). Compare punctate signal with adjacent cytoplasm and section morphology at the same magnification (standard IHC practice). Use compartment and cell identity together; membranous colour alone does not establish specific RAB11A staining (HPA: tissue IHC; standard IHC practice).
Can this IHC stain distinguish RAB11A isoforms or modified epitopes?
Do not assign staining to one isoform: RAB11A has 2 annotated isoforms, but the supplied antibody evidence does not identify its epitope (UniProt P62491 isoforms; caption M01436-2). The record also notes N terminal modification, a glycosylation site at residue 4, and a cysteine methyl ester at residue 213 (UniProt P62491 processing and modifications). Their effect on antibody binding has not been established for this IHC assay (caption M01436-2). Request the antibody's immunogen or epitope information before making isoform claims, and validate any proposed distinction with an independent method (standard IHC practice). Score the observed staining as RAB11A antibody reactivity until specificity is established (standard IHC practice).
How should I assess RAB11A by multiplex IF in tissue?
Treat IF conditions as a separate optimisation: the selected antibody image documents chromogenic paraffin IHC, while HPA reports vesicles and centriolar satellites in ICC/IF (caption M01436-2; HPA: subcellular). Multiplex with a marker for the expected cell population, such as an epithelial marker when examining HPA positive glandular cells, and confirm signal within those cells (HPA: tissue IHC; standard IF practice). Choose a spectrally separated, preferably far red fluorophore when tissue autofluorescence interferes, and include single stain controls (standard IF practice). RAB11A has no transmembrane segment, but the antibody epitope's orientation is unspecified; compare gentle permeabilisation conditions for access and preserved puncta (UniProt P62491 topology; standard IF practice).
How can I reduce diffuse brown staining without losing RAB11A puncta?
Check the no primary control, tissue edges and damaged areas before treating diffuse brown signal as RAB11A (standard IHC practice). The selected image used 10% goat serum blocking, a biotinylated secondary and a streptavidin biotin detection complex with DAB (caption M01436-2). With this detection chemistry, assess endogenous biotin and include a peroxidase block as general IHC controls (caption M01436-2; standard IHC practice). Titrate primary antibody around the documented 2 μg/ml condition and keep DAB development consistent when comparing sections (caption M01436-2; standard IHC practice). Preserve interpretable cytoplasmic or vesicular signal while reducing nonspecific deposition (UniProt P62491 localisation; standard IHC practice).
How should I quantify RAB11A IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HPA reports cytoplasmic and membranous expression across tissues (HPA: tissue IHC). For cell level scoring, record percent positive cells and staining intensity, or calculate an H score by summing intensity grades 0–3 weighted by their percentages (standard IHC practice). For punctate staining, report positive puncta or stained area per mm² of viable target tissue using a fixed image analysis threshold (standard IHC practice). Normalise counts to eligible cells or viable target area, and exclude folds, necrosis and tissue edges consistently (standard IHC practice). Compare sections processed with matching retrieval and detection conditions (datasheet M01436-2; standard IHC practice).
How do I separate true RAB11A signal from IHC artefact?
Look for reproducible cytoplasmic, vesicular or membranous staining in intact cells, consistent with reported RAB11A localisation (HPA: tissue IHC; UniProt P62491 localisation). Strong nuclear only staining, edge accentuation and colour confined to necrotic material call for control review (UniProt P62491 localisation; standard IHC practice). Check no primary and peroxidase controls when DAB deposits could reflect endogenous enzyme activity or detection background (caption M01436-2; standard IHC practice). The selected image shows staining in a human spleen section, whereas HPA reports red pulp cells as not detected; identify the stained cells before interpreting that difference (caption M01436-2; HPA: spleen red pulp). Confirm uncertain patterns with an independent antibody or orthogonal assay (standard IHC practice).
Boster reagents
Best RAB11A / Ras-related protein Rab-11A IHC Antibodies
Anti-RAB11A antibodies have IHC images from human paraffin sections and IF images from human cells (catalog image captions); all three list human, mouse and rat reactivity (catalog reactivity).
Real IHC data
Anti-Rab11A Antibody ® (monoclonal, 4H9)
Cat # M01436-2
Real IHC data
Anti-RAB11A Antibody
Cat # M01436-1
Real IF data
Anti-Rab11A Antibody ®
Cat # PB9789
M01436-2 has IHC images from human spleen, gastric carcinoma and placenta paraffin sections, plus IF data from T-47D cells (catalog image captions). M01436-1 has IHC-P data from formaldehyde-fixed human brain sections; PB9789 has IF/ICC data from U2OS cells (catalog image captions).
Which to pick: For tissue IHC, choose M01436-2 for human paraffin sections (M01436-2 IHC captions: EDTA retrieval; fixative unreported), or M01436-1 when paraffin-section paraffin sections match the workflow (M01436-1 IHC caption: citrate retrieval, 1:25). For IF/ICC, choose PB9789 or M01436-2 (catalog applications; respective U2OS and T-47D IF captions). All three list human, mouse and rat reactivity, but the supplied IHC images show human tissue only (catalog reactivity; IHC captions). The selected M01436-2 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image M01436-2).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P62491 (RB11A_HUMAN, Ras-related protein Rab-11A).
- Human Protein Atlas. RAB11A tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RAB11A subcellular location (ICC-IF): Mainly localized to vesicles and centriolar satellites. In addition localized to the cytosol, primary cilium, basal body, equatorial segment, mid piece and principal piece..
- Human Protein Atlas. RAB11A antibody validation summary (3 antibodies).
- Urinary exosomal RAB11A serves as a novel non-invasive biomarker for diagnosis, treatment response monitoring, and prognosis in small cell lung cancer. Clinical proteomics 2025 — PMC12376738.
- Recycling Endosomes in Mature Epithelia Restrain Tumorigenic Signaling. Cancer research 2019 — PMC6726494.
- Rab11a promotes proliferation and invasion through regulation of YAP in non-small cell lung cancer. Oncotarget 2017 — PMC5438609.
- Effects of miR‑340 overexpression and knockdown on the proliferation and metastasis of NSCLC cell lines. International journal of molecular medicine 2019 — PMC6605470.
- PubMed PMID:1704119 — UniProt-cited evidence.
- PubMed PMID:9662449 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RAB11A (Ras-related protein Rab-11A). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M01436-2). Validated IHC antibody: M01436-2 at 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human spleen tissue; fixative not specified (datasheet M01436-2); 2. Fixation: Image fixative and duration unreported (datasheet M01436-2); 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 M01436-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M01436-2); 8. Primary antibody: Mouse monoclonal (clone 4H9) anti-RAB11A, 2 μg/ml (datasheet M01436-2); 9. Primary incubation: Overnight at 4 °C (datasheet M01436-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Some cell populations, including adipocytes, show no signal (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage unknown; no transmembrane segment (UniProt); Regulation: No expression regulation annotated (UniProt). Evidence: UniProt P62491, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to vesicles and centriolar satellites. In addition…, Datasheet M01436-2, PMC12376738, PMC6726494. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RAB11A/immunohistochemistry-ihc-guide.