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GORAB / RAB6-interacting golgin · IHC design guide
Design Immunohistochemistry for GORAB
Plan GORAB paraffin-section IHC around its general cytoplasmic tissue staining (HPA tissue IHC). Colon glandular cells show high staining, while heart cardiomyocytes have no detected staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GORAB 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 in tissue (HPA tissue IHC); Golgi/TGN expected molecularly (UniProt) | |
| Staining pattern | General cytoplasmic staining in tissue cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A09417) | |
| Positive control | Cerebellum+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 | Medium consistency between staining and RNA expression (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | Two isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended GORAB IHC & IF Protocols
The catalog antibody protocol uses paraffin sections (datasheet A09417). One published mouse bone immunostaining protocol provides an additional IHC reference (PMC5880397).
| Sample | Paraffin-embedded human Colon cancer tissue; fixative not specified (datasheet A09417) |
| Fixation | Image fixative and duration unreported (datasheet A09417); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A09417); 20 min, 95–100 °C (standard) |
| 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-GORAB, 1:50 recommended; image 1:200 (datasheet A09417) |
| 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 | GORAB-positive staining in cells in granular layer of cerebellum (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 9.0 (datasheet A09417); the published protocol does not state a retrieval method (PMC5880397).
Section 2
What Is the Expected GORAB Staining Pattern?
GORAB localizes to the cytoplasm and trans-Golgi network (UniProt Q5T7V8). In tissue IHC, HPA reports general cytoplasmic staining, with high staining in selected neuronal, epithelial, glomerular and decidual cells (HPA tissue IHC). GORAB has no transmembrane segment (UniProt Q5T7V8 topology). Treat this as a working pattern: HPA rates its tissue IHC evidence Approved, with medium agreement between staining and RNA data, pending external verification (HPA tissue IHC).
| Cytoplasmic staining, sometimes concentrated near the nucleus, in the expected cell population. | Consistent with HPA's general cytoplasmic tissue pattern and UniProt's Golgi localization (HPA tissue IHC; UniProt Q5T7V8). A Golgi-like focus is plausible, but tissue IHC alone does not prove organelle identity. |
| Predominantly nuclear staining, or staining confined to cell borders or extracellular material. | Cell-border or extracellular staining is discordant with the reported cytoplasmic and Golgi pattern (HPA tissue IHC; UniProt Q5T7V8). Nuclear signal needs caution: HPA also reports nucleoplasm and nucleoli in ICC-IF, so nuclear staining alone does not establish an artefact (HPA subcellular ICC-IF). |
| Strong staining in cells expected to be unstained, such as adipocytes or cardiomyocytes. | These cells were not detected in HPA tissue IHC (HPA tissue IHC). Consider antibody cross-reactivity or endogenous chromogen activity; review morphology and detection controls before calling the cells GORAB-positive (general IHC practice). |
| Diffuse color across cells, stroma and empty spaces, with little cellular definition. | A broad, poorly localized signal cannot establish the reported cellular pattern (HPA tissue IHC). In chromogenic IHC, nonspecific binding, endogenous enzyme activity or detection background can produce this appearance (general IHC practice). |
| No staining in an expected positive cell population. | Absence in a high-staining HPA population, such as colon glandular cells, calls for a run-level check (HPA tissue IHC). Assess tissue preservation, retrieval and detection performance before interpreting the result as biological absence (general IHC practice). |
💡Expected GORAB appearanceA convincing positive shows discernible cytoplasmic staining, potentially with a Golgi-like focus, in an HPA high-staining cell population; widespread structureless color is suspect (HPA tissue IHC; UniProt Q5T7V8; general IHC practice).
| Tissue and cell selection | HPA reports high staining in colon and duodenal glandular cells, cerebral cortical neurons, cerebellar granular-layer cells, esophageal squamous epithelium, gallbladder glandular cells, glomerular cells and placental decidual cells (HPA tissue IHC). Compare the named cell population, not the whole section. |
| Negative comparison | HPA reports no detection in adipocytes, cardiomyocytes, skeletal myocytes, smooth muscle cells and splenic red-pulp cells (HPA tissue IHC). These are useful contextual comparisons, although absence in one sample cannot validate antibody specificity by itself (general IHC practice). |
| Localization evidence across applications | UniProt lists cytoplasm, Golgi apparatus and trans-Golgi network; HPA tissue IHC describes general cytoplasmic expression (UniProt Q5T7V8; HPA tissue IHC). HPA ICC-IF additionally reports nucleoplasm and nucleoli, so IF compartment calls should be assessed on its separate guide page (HPA subcellular ICC-IF). |
| Validation strength | HPA027250 is Approved for IHC, whereas the listed HPA027746 entry has no IHC status; both have Supported ICC status (HPA antibodies). HPA rates the tissue pattern Approved, with medium RNA agreement and external verification pending (HPA tissue IHC). |
| Protein features and epitope uncertainty | UniProt lists two isoforms, one 1–369 chain, no signal peptide, no propeptide and no annotated glycosylation sites or modified residues (UniProt Q5T7V8). The supplied record gives no antibody epitope, so isoform recognition and retrieval requirements cannot be inferred. |
| Situation | Likely cause | Next action |
|---|---|---|
| A high-staining reference population is blank. | The run may have inadequate retrieval, antibody binding or detection; HPA reports high staining in colon glandular cells (HPA tissue IHC; general IHC practice). | Check the reference slide, reagent sequence and chromogen performance; optimize retrieval against the antibody's validated IHC instructions (general IHC practice). |
| Expected cells stain, but the entire section has diffuse color. | Nonspecific binding or chromogenic detection background can obscure the cellular pattern (general IHC practice). | Review blocking, washes, detection timing and a no-primary control; score only cell-associated staining with clear morphology (general IHC practice). |
| Adipocytes or muscle cells appear strongly positive. | This conflicts with HPA's not-detected calls for adipocytes, cardiomyocytes, skeletal myocytes and smooth muscle cells (HPA tissue IHC). Cross-reactivity or endogenous activity is possible (general IHC practice). | Check cell identification and a no-primary control; reassess antibody and detection conditions before treating the signal as GORAB (general IHC practice). |
| Staining is mainly at cell borders or in extracellular material. | That distribution does not match the reported cytoplasmic and Golgi locations; UniProt reports no transmembrane segment (HPA tissue IHC; UniProt Q5T7V8 topology). | Compare with a reference population and detection control, then reassess staining localization before scoring it as positive (general IHC practice). |
| A predominantly nuclear signal conflicts with the expected tissue IHC pattern. | HPA tissue IHC describes general cytoplasmic staining, while HPA ICC-IF reports nucleoplasm as a main location (HPA tissue IHC; HPA subcellular ICC-IF). | Record nuclear and cytoplasmic staining separately. Confirm the tissue pattern with controls; use the separate IF/ICC guide for nuclear localization questions. |
| Different tissues or runs give inconsistent intensity. | HPA's Approved tissue pattern has medium agreement with RNA data and awaits external verification; intensity also depends on IHC run conditions (HPA tissue IHC; general IHC practice). | Compare the same named cell populations with a shared reference section and consistent scoring criteria across runs (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing GORAB staining
A09417 · IHCImmunohistochemical analysis of paraffin-embedded human Colon cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 w
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA027250; green = GORAB, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA027250; green = GORAB, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GORAB IHC & IF
🧪Run colon first: glandular cells should stain strongly (HPA: High in colon glandular cells). Use skeletal muscle as the negative tissue; myocytes are listed as not detected (HPA: Not detected in skeletal muscle myocytes). On the colon slide, compare glandular staining with surrounding nonglandular areas for background, without treating those cells as validated GORAB-negative cells.
Positive control tissue: Cerebellum (Cells in granular layer, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GORAB in A-431, U-251MG, U2OS, HAP1, SK-MEL-30, ASC52telo, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (approved), Golgi apparatus (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, host- and isotype-matched control immunoglobulin at the same concentration, and a GORAB knockout control or antigen-peptide competition if the relevant material is available (standard IHC practice). For colon chromogenic IHC, quench endogenous peroxidase and, if using avidin-biotin detection, block endogenous biotin (standard IHC practice).
⚠️Feasibility: No supplied source reports a target-specific fixation window or fixation effect, and the selected paraffin-section caption does not state the fixative (A09417 caption). The caption uses Tris-EDTA retrieval at pH 9.0 and primary antibody at 1:200 overnight at 4°C; it does not establish that retrieval is required (A09417 caption). Paraffin IHC has a documented starting condition, while the relative ease of frozen sections or IF is unreported; colon tissue autofluorescence can complicate IF interpretation (A09417 caption; standard IF practice).
HPA tissue IHC evidence for GORAB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GORAB IHC Tips
Troubleshoot GORAB staining in paraffin sections by checking retrieval, cell type, compartment and controls before comparing signal intensity (datasheet A09417; HPA tissue IHC).
Which retrieval should I use when GORAB staining is weak in paraffin sections?
Start with heat-mediated Tris-EDTA at pH 9.0 for antigen retrieval (datasheet A09417). The catalog antibody’s paraffin-section example used a 1:200 primary dilution overnight at 4°C, followed by a 1:200 secondary for 45 minutes at room temperature (caption A09417). Keep section thickness, heating conditions and detection constant while comparing retrieved and unretrieved sections (general IHC practice). Include colon glandular cells as a reference because high staining is reported there (HPA tissue IHC). If signal remains weak, troubleshoot antibody incubation and detection before interpreting the section as GORAB-negative (general IHC practice).
Can I infer the best fixative for GORAB from the paraffin-section example?
GORAB-specific fixation sensitivity is unknown: the catalog image describes paraffin-embedded colon cancer but does not state its fixative (caption A09417). Record the fixative and fixation duration for each specimen before comparing staining across samples (general IHC practice). When optimizing your workflow, hold retrieval at Tris-EDTA pH 9.0 and keep antibody incubation consistent while assessing independently prepared sections (datasheet A09417; general IHC practice). Examine preservation and staining in the same cell population, since damaged or poorly preserved regions can confound interpretation (general IHC practice). Neither the tissue staining profile nor GORAB topology establishes a preferred fixative (HPA tissue IHC; UniProt Q5T7V8).
Should chromogenic GORAB staining look diffuse or concentrated near the Golgi?
Assess cytoplasmic staining and any concentrated perinuclear component, since GORAB is annotated in the cytoplasm, Golgi apparatus and trans-Golgi network (UniProt Q5T7V8). The tissue IHC profile describes general cytoplasmic expression, so a visible Golgi focus is not required in every section (HPA tissue IHC). Compare the pattern within the expected cell population against adjacent background and a no-primary control (HPA tissue IHC; general IHC practice). Nuclear staining needs context: nucleoplasm and nucleoli are reported in cell-based localisation data, although those data do not establish a nuclear tissue-IHC pattern (HPA subcellular; HPA tissue IHC). Document compartment and intensity separately (general IHC practice).
How should I assess whether the antibody detects both GORAB isoforms?
GORAB has 2 annotated isoforms, but the supplied evidence does not identify the catalog antibody’s epitope or establish isoform coverage (UniProt Q5T7V8; caption A09417). Check the antibody’s epitope information, if available, against each isoform sequence before making an isoform-specific claim (general antibody-validation practice). GORAB has no annotated transmembrane segment, glycosylation sites or modified residues; these annotations do not identify an accessible antibody epitope in paraffin sections (UniProt Q5T7V8). If isoform recognition matters, compare staining with an independently validated antibody directed to a distinct known epitope and an appropriate specificity control (general IHC practice). Report the observed staining as GORAB staining unless isoform specificity is established (general IHC practice).
How can IF help resolve an ambiguous GORAB IHC pattern?
Use IF as a separate localisation check when chromogenic staining leaves the compartment unclear; this page’s documented tissue workflow is paraffin-section IHC (caption A09417; general microscopy practice). Multiplex GORAB with a marker for the expected cell population, such as an epithelial marker when examining colon glands, and compare signals cell by cell (HPA tissue IHC; general IF practice). Choose spectrally separated fluorophores and consider a far-red channel if tissue autofluorescence obscures shorter wavelengths (general IF practice). Because GORAB is intracellular and has no transmembrane segment, permeabilisation should permit antibody access to the intracellular epitope; the epitope itself is unspecified (UniProt Q5T7V8; general IF practice). Evaluate channel-specific controls before calling colocalisation (general IF practice).
What should I check when GORAB staining is widespread or granular?
First compare the slide with a no-primary control and inspect whether signal follows tissue edges, debris or endogenous pigment (general IHC practice). For peroxidase-based chromogenic detection, verify the peroxidase block and assess DAB development and counterstaining under the same conditions across slides (general IHC practice). Recheck blocking, wash steps and secondary-antibody background before increasing the primary concentration (general IHC practice). The catalog example used primary antibody at 1:200 overnight at 4°C and secondary antibody at 1:200 for 45 minutes (caption A09417). Interpret broad staining against the reported general cytoplasmic tissue pattern and cell-specific reference staining (HPA tissue IHC).
How should I score GORAB staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment and cell population before quantifying chromogenic GORAB signal (general IHC practice). For a selected population, report an H-score from 0–300 using intensity categories and the percentage of cells in each category, or report percent-positive cells with a stated threshold (general IHC practice). If spatial distribution matters, count positive cells per mm² of viable, annotated tissue and report the area denominator (general IHC practice). Normalize comparisons to the same cell type, viable area, staining batch and scoring rule; colon glandular cells provide a reported high-staining reference (HPA tissue IHC; general IHC practice). Record cytoplasmic and nuclear scores separately when both appear (HPA tissue IHC; HPA subcellular).
How do I distinguish credible GORAB staining from artefact?
A credible tissue-IHC result should be reproducible in the same cell population and judged against the reported general cytoplasmic pattern (HPA tissue IHC; general IHC practice). Colon glandular cells have reported high staining, whereas adipocytes are reported as not detected; use those observations as contextual references rather than absolute controls (HPA tissue IHC). Treat edge accentuation, necrotic regions and signal persisting without primary antibody as potential artefacts, including endogenous enzyme activity in peroxidase workflows (general IHC practice). A nuclear signal warrants separate review because cell-based data include nucleoplasm and nucleoli, while the tissue profile is generally cytoplasmic (HPA subcellular; HPA tissue IHC). Report unresolved compartment discrepancies explicitly (general IHC practice).
Boster reagents
Best GORAB / RAB6-interacting golgin IHC Antibodies
A09417 has a real IHC image from paraffin-embedded human colon cancer (catalog image caption). Human, mouse, and rat reactivity is listed (catalog reactivity); no IF image is supplied (catalog images).
Real IHC data
Anti-SCYL1BP1 GORAB Antibody
Cat # A09417
A09417 is the only card and shows IHC on paraffin-embedded human colon cancer, with 1:200 primary antibody dilution and Tris-EDTA retrieval at pH 9.0 (catalog image caption). Its listed applications include IHC, and its listed reactivity covers human, mouse, and rat (catalog applications/reactivity).
Which to pick: Choose A09417, a rabbit polyclonal, for paraffin-section tissue IHC because its own image documents that preparation; the fixative is unreported (catalog dilution data; catalog image caption). No SKU has listed IF/ICC application or IF image data here (catalog applications/images). For mouse or rat IHC, A09417 lists those species as reactive, but its supplied IHC image shows human tissue only (catalog reactivity; catalog image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q5T7V8 (GORAB_HUMAN, RAB6-interacting golgin).
- Human Protein Atlas. GORAB tissue IHC expression (reliability: Approved).
- Human Protein Atlas. GORAB subcellular location (ICC-IF): Mainly localized to the nucleoplasm and the Golgi apparatus. In addition localized to the nucleoli and cytosol..
- Human Protein Atlas. GORAB antibody validation summary (2 antibodies).
- Gorab deficiency in skin dermis accelerates aging and is associated with dysregulation of RCHY1-mediated P53 ubiquitination. Animal models and experimental medicine 2026 — PMC13176089.
- Impaired proteoglycan glycosylation, elevated TGF-β signaling, and abnormal osteoblast differentiation as the basis for bone fragility in a mouse model for gerodermia osteodysplastica. PLoS genetics 2018 — PMC5880397.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GORAB (RAB6-interacting golgin). Expected localisation Cytoplasmic in tissue (HPA tissue IHC); Golgi/TGN expected molecularly (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A09417). Validated IHC antibody: A09417 at 1:50 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Cells in granular layer); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Colon cancer tissue; fixative not specified (datasheet A09417); 2. Fixation: Image fixative and duration unreported (datasheet A09417); 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: Tris-EDTA pH 9.0 (datasheet A09417); 20 min, 95–100 °C (standard); 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-GORAB, 1:50 recommended; image 1:200 (datasheet A09417); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Medium consistency between staining and RNA expression (HPA tissue IHC); Isoform / epitope: Two isoforms; epitope coverage is unknown (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q5T7V8, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and the Golgi apparatus. In…, Datasheet A09417, PMC5880397. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GORAB/immunohistochemistry-ihc-guide.