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GORASP2 / Golgi reassembly-stacking protein 2 · IHC design guide
Design Immunohistochemistry for GORASP2
Plan chromogenic IHC for GORASP2 using the reported cytoplasmic and membranous tissue pattern, including strong staining in gastrointestinal endocrine cells (HPA tissue IHC). This guide covers fixation, antibody dilution, controls and scoring, while accounting for the reported mismatch between staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GORASP2 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 | Cytoplasmic/membranous staining in GI endocrine cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A07850-1) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Caudate+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 A07850-1) | |
| Caveat | Staining may diverge from RNA abundance (HPA tissue IHC) | |
| Regulation | Intensity regulation unreported (UniProt) | |
| Isoform / epitope | 3 isoforms; confirm epitope coverage (UniProt) |
Section 1
Recommended GORASP2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published GORASP2 protocol using FFPE sections (PMC6422190).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A07850-1) |
| Fixation | Image fixative and duration unreported (datasheet A07850-1); 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 A07850-1); 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-GORASP2, 1:100-1:300 (datasheet A07850-1) |
| 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 | GORASP2-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in several tissues, mainly in the gastrointestinal tract. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA pH 9.0 for the catalog antibody (datasheet A07850-1); use citrate pH 6.0 when reproducing the published protocol (PMC6422190).
Section 2
What Is the Expected GORASP2 Staining Pattern?
GORASP2 is a Golgi-associated protein with no transmembrane segment; ER stress can relocate it to the ER membrane (UniProt Q9H8Y8). In paraffin-section IHC, expect cytoplasmic and membranous staining in selected cells, including gastrointestinal endocrine cells and respiratory epithelial cells (HPA tissue IHC). HPA rates the tissue staining evidence Enhanced, while noting low consistency with RNA expression; paired antibodies support the protein pattern (HPA tissue IHC).
| Localized cytoplasmic staining in endocrine cells of appendix, colon, or duodenum, with little staining in adjacent cells. | This fits the reported High staining in those endocrine cell populations (HPA tissue IHC). A compact, perinuclear distribution is compatible with Golgi localization (UniProt Q9H8Y8; HPA subcellular ICC-IF). Judge the named cell population, since HPA does not assign one intensity to every cell in the section (HPA tissue IHC). |
| Predominantly nuclear, extracellular, or uniform cell-surface staining replaces the expected cytoplasmic pattern. | These compartments do not match the reported Golgi localization or tissue-level cytoplasmic and membranous profile (HPA subcellular ICC-IF; HPA tissue IHC). Treat the result as suspect and review controls and section morphology (general IHC practice). The tissue IHC profile alone cannot resolve every intracellular membrane. |
| Strong staining appears in glomerular cells or liver cholangiocytes. | HPA reports these specific cell populations as Not detected (HPA tissue IHC). Reproducible staining warrants checking antibody specificity and endogenous detection activity (general IHC practice). Do not extend those cell-specific negative observations to all kidney or liver cells (HPA tissue IHC). |
| Color is diffuse across tissue structures, including areas without plausible cellular localization. | A broadly distributed deposit is hard to reconcile with the reported Golgi-centered localization (HPA subcellular ICC-IF). Background can arise from the detection workflow, inadequate blocking, or excess antibody (general IHC practice); the appearance alone does not identify which step is responsible. |
| No staining is visible in a well-preserved appendix, colon, or duodenum endocrine cell population. | Those populations are reported High by HPA, so absent signal calls for a technical review (HPA tissue IHC; general IHC practice). Confirm the cells are present and examine retrieval, antibody use, and detection controls (general IHC practice). HPA tissue levels do not establish a GORASP2-specific fixation sensitivity. |
💡Expected GORASP2 appearanceCall a result positive when selected epithelial or endocrine cells show discernible cytoplasmic, Golgi-compatible staining, potentially High in the named HPA-positive populations; widespread nuclear or structure-independent color is suspect (HPA tissue IHC; HPA subcellular ICC-IF; general IHC practice).
| Cell selection in the section | HPA reports High staining in respiratory epithelial cells of bronchus and nasopharynx, and glandular cells of endometrium, epididymis, and prostate, alongside the gastrointestinal endocrine populations (HPA tissue IHC). Score the identified cells rather than treating the entire organ as uniformly positive (general IHC practice). |
| Reliability of the tissue pattern | The IHC profile is Enhanced because paired antibodies show similar protein staining, despite low agreement with RNA expression (HPA tissue IHC). This supports the observed pattern but does not make each positive or negative cell assignment an absolute biological rule. |
| Intracellular location | UniProt places GORASP2 at the Golgi and describes it as membrane-associated, with no transmembrane segment (UniProt Q9H8Y8). HPA reports a Golgi main location by ICC-IF (HPA subcellular ICC-IF). A tissue-level description of membranous staining should therefore not automatically be scored as plasma-membrane staining. |
| ER stress | ER stress can relocalize GORASP2 from the Golgi to the ER membrane (UniProt Q9H8Y8). A less compact cytoplasmic distribution may merit biological review when ER stress is part of the experiment; the supplied HPA tissue images do not establish how that change appears in chromogenic paraffin-section IHC. |
| IF/ICC: where should the signal appear? | HPA reports enhanced Golgi localization in ICC-IF images, including U2OS cells with siRNA and scrambled conditions (HPA subcellular ICC-IF). Use that as a compartment cross-check; ICC-IF images do not specify an IHC-P retrieval or staining protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control endocrine cells are blank. | The target cells may be absent from the examined level, or the IHC workflow may have failed (HPA tissue IHC; general IHC practice). | Verify cell identity and preservation, then review antigen retrieval, primary antibody application, and detection controls (general IHC practice). Do not infer a GORASP2-specific fixation effect from a blank section. |
| All structures carry a pale, even chromogen deposit. | Nonlocalized signal can reflect background from blocking, washing, antibody concentration, or detection chemistry (general IHC practice). | Inspect the negative reagent control, wash quality, blocking, and chromogen development; adjust one workflow variable at a time (general IHC practice). |
| Staining appears in glomerular cells or cholangiocytes. | These cells are reported Not detected by HPA; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Compare a negative reagent control and an appropriate positive-cell control, and confirm the stained cell type before interpreting the result (general IHC practice). |
| The strongest signal is nuclear. | A nuclear-dominant pattern conflicts with the Golgi localization reported by UniProt and HPA (UniProt Q9H8Y8; HPA subcellular ICC-IF). | Recheck counterstain, chromogen distribution, and controls; report the compartment mismatch rather than scoring it as expected GORASP2 staining (general IHC practice). |
| A presumed negative section contains a few stained cells. | HPA negative calls apply to named cells, such as red-pulp cells in spleen, rather than every cell in an organ (HPA tissue IHC). | Identify the stained cells first, then compare that exact cell population with the HPA entry and the negative reagent control (HPA tissue IHC; general IHC practice). |
| Cytoplasmic staining is broad instead of compact and perinuclear. | ER stress can redistribute GORASP2 to the ER membrane, while nonspecific background remains another possibility (UniProt Q9H8Y8; general IHC practice). | Check whether ER stress is relevant to the specimen, then compare cellular boundaries and negative controls; do not assign ER redistribution from chromogenic appearance alone (UniProt Q9H8Y8; general IHC practice). |
Experiment evidence
IHC & IF images showing GORASP2 staining
A07850-1 · IHCImmunohistochemical analysis of paraffin-embedded human liver cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 w
A30705 · IHCImmunohistochemical analysis of paraffin-embedded human liver cancer. 1, Antibody was diluted at 1:200(4° overnight). 2, Tris-EDTA,pH9.0 was
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115806-GORASP2/tissue/cerebral+cortex#imid_19814189
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115806-GORASP2/tissue/colon#imid_19814123
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115806-GORASP2/tissue/liver#imid_19814116
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000115806-GORASP2/tissue/kidney#imid_19814125
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035274; green = GORASP2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035274; green = GORASP2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GORASP2 IHC & IF
🧪Run appendix first and require staining in its endocrine cells (HPA: High in appendix endocrine cells); use caudate glial cells as a negative comparator (HPA: Not detected in caudate glial cells). Adjacent appendix cells can serve as internal negatives only if they show background chromogen without a discrete Golgi pattern; their negative status is not established by the supplied HPA rows (HPA subcellular: Golgi apparatus enhanced).
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GORASP2 in A-431, U-251MG, U2OS, U2OS, siRNA 1 (10x), U2OS, siRNA 2 (10x), U2OS, scrambled (10x), with annotated localisation: Golgi apparatus (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species-matched isotype controls for a monoclonal primary, or matched normal IgG for a polyclonal primary, plus GORASP2-knockout material where available as a biological negative (standard IHC practice). For chromogenic appendix IHC, block endogenous peroxidase and assess any nonspecific tissue pigment against the no-primary slide (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 (selected-SKU caption: fixative not stated). The caption uses Tris-EDTA at pH 9.0 for retrieval and primary antibody at 1:200 overnight at 4°C; it does not establish that retrieval is required (selected-SKU caption: retrieval and dilution). Is frozen section or IF easier? The supplied evidence does not establish that either is easier; IF/ICC can assess the expected Golgi localization, and no appendix-specific artefact is reported (HPA subcellular: Golgi apparatus enhanced; supplied HPA tissue rows).
HPA tissue IHC evidence for GORASP2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Low consistency between antibody staining and RNA expression data. Paired antibodies with high similarity supports the protein expression.). 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 |
|---|---|---|---|---|
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GORASP2 IHC Tips
Troubleshoot GORASP2 chromogenic IHC by checking retrieval, tissue preservation, staining pattern, controls, and scoring before interpreting biological differences.
What retrieval should I start with when GORASP2 staining is weak?
Start with heat-mediated Tris-EDTA at pH 9.0 for paraffin sections (datasheet A07850-1). The selected tissue image used this retrieval with primary antibody at 1:200 overnight at 4°C, so reproduce those documented conditions before changing several variables together (caption A07850-1). If staining remains weak, compare a shorter and a longer heating exposure on adjacent sections while keeping detection and imaging settings constant; inspect morphology after each run. Include a positive control with high endocrine-cell staining in appendix or colon, and assess those cells rather than whole-section color (HPA tissue IHC: High in appendix and colon endocrine cells).
How should I troubleshoot suspected fixation-related loss of GORASP2 staining?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not report a fixative (caption A07850-1). Record each specimen’s fixative, time in fixative, processing history, and section age before attributing weak staining to fixation. Compare an affected section with a similarly processed positive control, keeping Tris-EDTA at pH 9.0 and the documented 1:200 primary dilution constant (datasheet A07850-1; caption A07850-1). If staining differs across processing batches, repeat the comparison with matched tissue sections and review morphology; neither tissue staining patterns nor protein topology establishes a GORASP2-specific fixation effect.
Where should GORASP2 signal appear in chromogenic tissue sections?
Expect a predominantly perinuclear, Golgi-region cytoplasmic pattern where cellular detail is preserved: the protein localizes to the Golgi apparatus (HPA subcellular: Golgi apparatus, enhanced; UniProt Q9H8Y8 localisation). Tissue IHC also reports cytoplasmic and membranous staining, especially in gastrointestinal tissues, so interpret a broader pattern against cell morphology and matched controls (HPA tissue IHC profile). UniProt also lists ER membrane localization and ER stress-associated relocalization, which cannot be established from a single chromogenic section (UniProt Q9H8Y8 localisation). If staining is uniformly nuclear or diffuse across every compartment, check background and detection controls before treating it as a new distribution.
Could isoforms or modifications explain discordant GORASP2 IHC staining?
GORASP2 has 3 annotated isoforms and two GRASP-type PDZ domains spanning residues 15–105 and 111–199 (UniProt Q9H8Y8 isoforms and domains). Its annotated modifications include arginine dimethylation and phosphorylation, but the supplied caption does not identify this antibody’s epitope (UniProt Q9H8Y8 modified residues; caption A07850-1). Ask for the immunogen or mapped epitope before claiming isoform-selective staining or modification sensitivity. When two antibodies disagree, compare their documented epitopes, retrieval conditions, and staining in the same cell population; use an independent specificity control before assigning the difference to an isoform.
How can IF clarify an ambiguous GORASP2 IHC pattern?
Use IF as a complementary localization check: GORASP2 has enhanced Golgi localization in cell imaging, while tissue IHC reports cytoplasmic and membranous staining (HPA subcellular; HPA tissue IHC profile). Multiplex with a validated marker of the expected cell type, such as an endocrine-cell marker when examining appendix or colon endocrine cells, and evaluate overlap within individual cells (HPA tissue IHC: High in appendix and colon endocrine cells). Choose fluorophores and acquisition channels after measuring unstained-tissue autofluorescence, and include single-stain controls. GORASP2 is membrane-associated without a transmembrane segment, so optimize permeabilisation to expose an intracellular epitope once the antibody’s epitope and its membrane-facing side are established (UniProt Q9H8Y8 topology and localisation).
How do I reduce diffuse brown background without losing genuine signal?
First inspect a no-primary control and the tissue edges to separate detection background from cell-associated staining; the expected signal has a Golgi-centered component (HPA subcellular: Golgi apparatus, enhanced). In a peroxidase-based chromogenic workflow, check the peroxide block and review DAB development time as general IHC controls. Titrate blocking, washes, and primary concentration around the documented 1:200 dilution while keeping retrieval at Tris-EDTA pH 9.0 (caption A07850-1; datasheet A07850-1). Compare candidate settings in the same staining run with a high-staining reference cell population and a low-staining comparator, judging cellular pattern as well as overall darkness (HPA tissue IHC).
How should I score GORASP2 staining across heterogeneous sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; tissue IHC reports cytoplasmic and membranous expression, while cell imaging places GORASP2 mainly at the Golgi (HPA tissue IHC profile; HPA subcellular). For a chromogenic intensity readout, report the percentage of positive cells and an H-score from intensity grades 0–3, with a stated positivity threshold. If the question concerns stained-cell abundance, report positive-cell density per mm² of viable, annotated tissue. Normalize comparisons to the same eligible cell population and tissue area, exclude necrosis and section edges by prespecified rules, and hold staining and image-analysis settings constant across batches.
What distinguishes true GORASP2 staining from an IHC artefact?
A credible positive should follow cell boundaries and show a plausible cytoplasmic, often perinuclear Golgi-region pattern (HPA tissue IHC profile; HPA subcellular: Golgi apparatus, enhanced). Check the expected cell type: endocrine cells in appendix and colon are reported as high, whereas liver cholangiocytes are reported as not detected; neither observation validates every cell in a specimen (HPA tissue IHC). Treat staining confined to cut edges, necrotic regions, nuclei, or every tissue element as suspect until controls support it. Compare no-primary and peroxidase-block controls, then repeat the finding in intact, morphologically identifiable cells before interpreting intensity differences as biological.
Boster reagents
Best GORASP2 / Golgi reassembly-stacking protein 2 IHC Antibodies
Two anti-GORASP2 antibodies have IHC images from paraffin-embedded human liver cancer (each SKU’s IHC caption); both list IF and human, mouse and rat reactivity (catalog applications and reactivity).
Real IHC data
Anti-GRASP55 GORASP2 Antibody
Cat # A07850-1
Real IHC data
Anti-GORASP2/Grasp55 Antibody
Cat # A30705
A07850-1 lists IHC and IF for human, mouse and rat, with an IHC image from paraffin-embedded human liver cancer (catalog applications and reactivity; A07850-1 IHC caption). A30705 lists IHC, IF and ICC for human, mouse and rat, with its own IHC image from paraffin-embedded human liver cancer (catalog applications and reactivity; A30705 IHC caption).
Which to pick: For tissue IHC, either SKU has an image from paraffin-embedded human liver cancer; each caption reports 1:200 primary antibody and Tris-EDTA retrieval at pH 9.0, but neither reports the fixative (each SKU’s IHC caption). For IF/ICC, choose A30705 because both applications are listed; A07850-1 lists IF without ICC (catalog applications). For mouse or rat samples, both list reactivity, but the supplied IHC images show human tissue only (catalog reactivity; each SKU’s IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H8Y8 (GORS2_HUMAN, Golgi reassembly-stacking protein 2).
- Human Protein Atlas. GORASP2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GORASP2 subcellular location (ICC-IF): Localized to the Golgi apparatus..
- Human Protein Atlas. GORASP2 antibody validation summary (3 antibodies).
- Proper acquisition of cell class identity in organoids allows definition of fate specification programs of the human cerebral cortex. Cell 2022 — PMC9990683.
- DNA hypomethylation-related overexpression of SFN, GORASP2 and ZYG11A is a novel prognostic biomarker for early stage lung adenocarcinoma. Oncotarget 2019 — PMC6422190.
- PubMed PMID:11101516 — UniProt-cited evidence.
- PubMed PMID:11408587 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GORASP2 (Golgi reassembly-stacking protein 2). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A07850-1). Validated IHC antibody: A07850-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Caudate. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A07850-1); 2. Fixation: Image fixative and duration unreported (datasheet A07850-1); 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 retrieval: Tris-EDTA pH 9.0 (datasheet A07850-1); 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-GORASP2, 1:100-1:300 (datasheet A07850-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07850-1); Caveat: Staining may diverge from RNA abundance (HPA tissue IHC); Isoform / epitope: 3 isoforms; confirm epitope coverage (UniProt); Regulation: Intensity regulation unreported (UniProt). Evidence: UniProt Q9H8Y8, HPA tissue IHC · Enhanced, HPA ICC-IF · Golgi apparatus., Datasheet A07850-1, PMC6422190. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GORASP2/immunohistochemistry-ihc-guide.