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GTPBP2 / GTP-binding protein 2 · IHC design guide
Design Immunohistochemistry for GTPBP2
Plan GTPBP2 staining in paraffin sections around the reported cytoplasmic tissue pattern (HPA tissue IHC). The catalog antibody has an IHC dilution range of 1:100–1:300 (datasheet); appendix glandular cells show medium staining, while adipocytes are undetected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GTPBP2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in glandular and other cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| 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 paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Spleen red pulp lacks staining despite reported spleen expression (HPA tissue IHC; UniProt) | |
| Regulation | Tissue-dependent expression; no regulator specified (UniProt) | |
| Isoform / epitope | 4 isoforms; epitope coverage is unresolved (UniProt; datasheet) |
Section 1
Recommended GTPBP2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published chromogenic IHC methods for lung cancer specimens (PMC8176249) and mouse colorectal cancer tissue sections (PMC10926081).
| Sample | Paraffin-embedded human colon carcinoma tissue; fixative not specified (datasheet A30713) |
| Fixation | Image fixative and duration unreported (datasheet A30713); 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-GTPBP2, 1:100 - 1:300 (datasheet A30713) |
| 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 | GTPBP2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen-retrieval rule). The lung cancer study reports high-pressure citrate retrieval for 2 min (PMC8176249).
Section 2
What Is the Expected GTPBP2 Staining Pattern?
GTPBP2 IHC should show predominantly cytoplasmic staining in many tissues, including medium staining in adrenal and appendix glandular cells, cerebellar Purkinje cells, and colon endothelial cells (HPA tissue IHC: Approved; medium consistency with RNA data, pending external verification). UniProt does not assign a subcellular location and reports no transmembrane segment (UniProt Q9BX10 topology); the slide pattern is grounded in HPA observations.
| Cytoplasmic color in adrenal or appendix glandular cells, with identifiable cell boundaries (HPA tissue IHC: Medium in these cells). | This fits the reported IHC pattern (HPA tissue IHC: cytoplasmic expression in most tissues). Judge the labeled cells and compartment together; medium staining is an observed level, not a required intensity for every specimen (HPA tissue IHC: Approved, with medium consistency). |
| Color confined mainly to nuclei, membranes, or extracellular material instead of cytoplasm. | This departs from the reported tissue IHC distribution (HPA tissue IHC: cytoplasmic expression). Treat it as possible artefact and check controls and tissue morphology before calling it GTPBP2; lack of a transmembrane segment alone cannot prove a particular microscopic location (UniProt Q9BX10 topology). |
| Strong color in adipocytes or cervical squamous epithelial cells while expected positive cells stain (HPA tissue IHC: Not detected in those cells). | Question cell identification, cross-reactivity, and endogenous chromogenic activity (standard IHC practice). HPA's negative calls apply to the specified cell populations, not automatically to every cell in those tissues (HPA tissue IHC: cell-level observations). |
| Diffuse color covers several compartments or obscures the boundary between cells. | That appearance cannot establish the reported cytoplasmic pattern (HPA tissue IHC: cytoplasmic expression). Assess a reagent omission control and detection background, then adjust blocking, washing, or antibody concentration as appropriate (standard IHC practice). |
| No color in adrenal glandular cells or cerebellar Purkinje cells (HPA tissue IHC: Medium in these cells). | First suspect an assay or tissue-control failure; verify tissue identity, detection reagents, and the validated IHC antibody (standard IHC practice). A single unstained specimen does not overturn an HPA reference pattern, particularly given its medium consistency and pending external verification (HPA tissue IHC: reliability description). |
💡Expected GTPBP2 appearanceCall a positive result when recognizable glandular cells, Purkinje cells, or endothelial cells show mainly cytoplasmic color at the reported medium level; isolated nuclear or extracellular color, or strong staining in HPA-negative cell populations, is suspect (HPA tissue IHC: positive and negative cell-level observations; cytoplasmic profile).
| Cell population and tissue | The sampled cells determine what can be scored: adrenal and appendix glandular cells, Purkinje cells, and colon endothelial cells have Medium calls, whereas adipocytes and spleen red-pulp cells are Not detected (HPA tissue IHC). |
| RNA and protein context | UniProt reports predominant expression in thymus, spleen, and testis, while HPA reports Not detected specifically in spleen red-pulp cells (UniProt Q9BX10 tissue specificity; HPA tissue IHC). Do not turn either statement into a whole-spleen IHC prediction. |
| Validation and compartment evidence | The tissue IHC profile is Approved, with medium antibody–RNA consistency and external verification pending; HPA031419 has Approved IHC status (HPA tissue IHC; HPA antibodies). UniProt gives no subcellular annotation, so the cytoplasmic call rests on HPA tissue staining (UniProt Q9BX10; HPA tissue IHC). |
| Isoforms and antibody recognition | Four isoforms are listed (UniProt Q9BX10). Their relative staining, epitopes, and tissue distribution are not supplied; an isoform-specific interpretation would need separate evidence (UniProt Q9BX10 isoform record). |
| IF/ICC Q&A: should puncta match IHC? | HPA reports an approved vesicular ICC-IF location, with images from A-431, HeLa, and U2OS, using ICC-approved HPA031418 (HPA subcellular; HPA antibodies). Tissue IHC is summarized as cytoplasmic and has Approved status for HPA031419; interpret the methods and antibodies separately (HPA tissue IHC; HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells have no chromogenic signal. | Tissue identification, primary antibody, retrieval, or detection may have failed (standard IHC practice); HPA reports Medium staining in adrenal glandular cells (HPA tissue IHC). | Confirm the cell population and positive control, then check the IHC-validated antibody, detection reagents, and retrieval conditions (standard IHC practice). No GTPBP2-specific retrieval or fixation effect is supplied (UniProt Q9BX10; HPA tissue IHC). |
| Background appears across the section. | Nonspecific reagent binding, incomplete washing, or detection background can obscure cell boundaries (standard IHC practice). | Compare a primary-antibody omission control; review blocking, wash conditions, and antibody concentration before scoring cytoplasm (standard IHC practice; HPA tissue IHC: cytoplasmic profile). |
| Color is concentrated in nuclei or extracellular spaces. | The location conflicts with the reported tissue pattern (HPA tissue IHC: cytoplasmic expression); artefact or misread morphology is possible (standard IHC practice). | Inspect morphology and controls, and score only recognizable cells with a credible cytoplasmic pattern (standard IHC practice; HPA tissue IHC). |
| Unexpected cells stain strongly. | Cross-reactivity or endogenous chromogenic activity is possible (standard IHC practice); adipocytes and cervical squamous cells are Not detected in the HPA observations (HPA tissue IHC). | Check the exact cell type and a reagent omission control; reassess specificity before treating that color as GTPBP2 (standard IHC practice; HPA tissue IHC: cell-level negatives). |
| Spleen results seem to conflict with expression data. | UniProt lists predominant spleen expression, but HPA's negative IHC observation names red-pulp cells specifically (UniProt Q9BX10 tissue specificity; HPA tissue IHC). | Record which splenic cells were scored and keep the RNA or tissue-level statement separate from the red-pulp IHC observation (UniProt Q9BX10; HPA tissue IHC). |
| IF/ICC shows vesicular puncta while IHC looks broadly cytoplasmic. | HPA reports vesicles for ICC-IF and a cytoplasmic profile for tissue IHC; the approved antibodies also differ (HPA subcellular; HPA tissue IHC; HPA antibodies). | Document the method and antibody identifier, then interpret each against its matching HPA reference rather than requiring identical appearance (HPA antibodies: HPA031418 ICC Approved; HPA031419 IHC Approved). |
Experiment evidence
IHC & IF images showing GTPBP2 staining
A30713 · IHCImmunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue, using GTPBP2 Antibody. The picture on the right is blocked
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000172432-GTPBP2/tissue/cerebral+cortex#imid_8413372
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000172432-GTPBP2/tissue/colon#imid_8413282
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000172432-GTPBP2/tissue/liver#imid_8413283
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000172432-GTPBP2/tissue/kidney#imid_8413284
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031418; green = GTPBP2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031418; green = GTPBP2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GTPBP2 IHC & IF
🧪Run cerebellum first and look for GTPBP2 staining in Purkinje cells (HPA: Medium in cerebellar Purkinje cells). Use hippocampus as a negative comparator, scoring its glial cells (HPA: Not detected in hippocampal glial cells); on the cerebellar slide, non-Purkinje cells that lack signal above background can serve as internal comparators, but their GTPBP2 status is not established by the supplied HPA rows.
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 GTPBP2 in A-431, HeLa, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a control immunoglobulin matched to the primary antibody’s host species, isotype and monoclonal or polyclonal format (standard IHC practice). A GTPBP2 knockout specimen or cognate-peptide block provides a biological specificity check (A30713 caption: staining blocked with synthesized peptide); quench endogenous peroxidase and compare cerebellar neuronal pigment with the no-primary slide during chromogenic scoring (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 (A30713 caption: fixative unreported). Retrieval dependence is unreported, so optimize antigen retrieval for the IHC-validated antibody; the supplied evidence does not establish whether frozen sections or IF are easier, although vesicular ICC-IF localization is reported in A-431, HeLa and U2OS cells (HPA: Vesicles, approved). Neuronal lipofuscin in cerebellum can resemble chromogen and autofluoresce, so check the no-primary control when scoring Purkinje cells (standard histology practice).
HPA tissue IHC evidence for GTPBP2
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GTPBP2 IHC Tips
Troubleshoot GTPBP2 staining in paraffin sections by checking retrieval, compartment, cell type, and controls before interpreting chromogenic signal.
How should I adjust retrieval when GTPBP2 staining is weak in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 minutes (page retrieval rule). Let sections cool in buffer, then compare a retrieved section with an unretrieved section using the same detection conditions (standard IHC practice). The selected antibody has a paraffin-section image, but its caption does not report retrieval conditions or fixative (caption: A30713). If staining remains weak, test a more alkaline retrieval buffer on adjacent sections as a fallback, watching for tissue damage and rising background (standard IHC practice). Keep the peptide-blocked image as evidence for the illustrated staining, not as proof of the best retrieval method (caption: A30713).
Could fixation explain inconsistent GTPBP2 staining between paraffin blocks?
GTPBP2-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (caption: A30713). Compare blocks with documented processing histories and stain sections together so retrieval and detection vary as little as possible (standard IHC practice). Excessive or uneven fixation can alter epitope access in paraffin IHC generally, but it cannot be assigned as the cause here without a controlled comparison (standard IHC practice). If archival blocks differ, include a shared reference section in each staining run and record staining intensity alongside fixation records (standard IHC practice). Do not use tissue-expression patterns to infer fixation sensitivity (HPA: tissue IHC profile).
Where should convincing GTPBP2 chromogenic signal appear?
Expect predominantly cytoplasmic tissue staining, while recognizing that the separate cell-imaging record places GTPBP2 in vesicles (HPA: tissue IHC profile; HPA: subcellular localisation). The protein has no annotated transmembrane segment, and UniProt supplies no subcellular localisation assignment (UniProt Q9BX10: topology and localisation). Assess signal within intact cells under a nuclear counterstain, separating cytoplasmic color from pigment, extracellular deposits, and section folds (standard IHC practice). Medium staining is reported in colon endothelial cells, providing a specified cell population to inspect on a comparison section (HPA: colon tissue IHC). A strong nuclear-only pattern warrants reassessment with controls before being called GTPBP2 positive (HPA: tissue IHC profile).
Can this antibody distinguish GTPBP2 isoforms in paraffin sections?
UniProt lists 4 GTPBP2 isoforms, but the supplied antibody caption does not identify its epitope or establish isoform selectivity (UniProt Q9BX10: isoforms; caption: A30713). The tr-type G domain spans residues 170–398; domain annotation alone does not show where this antibody binds (UniProt Q9BX10: domains). Check an available epitope map against each isoform before claiming isoform-specific staining (standard IHC interpretation). The caption shows peptide blocking in a human colon carcinoma paraffin section, which supports competition of the illustrated signal under those conditions (caption: A30713). It does not establish which isoform produced that signal or exclude cross-reactivity elsewhere (caption: A30713).
How can IF help assess the cellular source of GTPBP2 IHC staining?
For the separate IF assay, pair GTPBP2 with an endothelial marker when investigating colon staining, because HPA reports medium GTPBP2 staining in colon endothelial cells (HPA: colon tissue IHC). Choose a far-red fluorophore when green tissue autofluorescence obscures puncta, and acquire single-label controls to check spectral bleed-through (standard IF practice). The cell-imaging record reports vesicular localisation, so use mild permeabilisation to access intracellular epitopes, then compare permeabilised and untreated specimens because this antibody’s epitope is unspecified (HPA: subcellular localisation; standard IF practice). Include secondary-only controls and inspect whether signal overlaps the expected cell population rather than relying on color alone (standard IF practice). Do not transfer the paraffin-section caption’s unstated fixation conditions to IF (caption: A30713).
What should I check when GTPBP2 chromogenic staining looks diffuse?
First inspect a no-primary control for endogenous enzyme signal and nonspecific detection, then compare it with the GTPBP2-stained section (standard IHC practice). Apply a peroxidase block before an HRP and DAB workflow, and keep DAB development time consistent across sections (standard IHC practice). Check whether brown color accumulates at section edges, folds, necrotic areas, or pigment rather than within intact cytoplasm (standard IHC practice; HPA: tissue IHC profile). If background rises after retrieval, compare adjacent sections retrieved for 20 minutes under the page condition and a shorter trial exposure (page retrieval rule; standard IHC practice). The peptide-blocked product image is a useful reference for competition, but it does not replace run-specific negative controls (caption: A30713).
How should I score GTPBP2 across paraffin sections with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring; HPA describes broadly cytoplasmic tissue staining and specifically reports medium staining in colon endothelial cells (HPA: tissue IHC profile; HPA: colon tissue IHC). For a defined population, record the percentage of positive cells and intensity grades 0–3, then calculate an H-score on a 0–300 scale (standard IHC scoring practice). Where cell abundance varies, report positive cells per mm² of viable tissue or normalize to the number of eligible cells (standard IHC scoring practice). Exclude folds, necrosis, and damaged edges using criteria set before review (standard IHC practice). Keep retrieval, counterstain, illumination, and scoring thresholds consistent between groups (standard IHC practice).
Which observations support genuine GTPBP2 staining rather than artefact?
Give greatest weight to reproducible intracellular staining in intact cells that agrees with the predominantly cytoplasmic tissue profile (HPA: tissue IHC profile). Colon endothelial cells offer one specified comparison population, while HPA reports no detection in adipocytes and ovarian stroma cells in its sampled tissues (HPA: colon, adipose tissue, and ovary IHC). A nuclear-only pattern, color confined to section edges or necrosis, or staining reproduced in a no-primary control calls for investigation (HPA: tissue IHC profile; standard IHC practice). The peptide-blocked paraffin image supports competition of its illustrated colon carcinoma signal, but its fixative is unreported (caption: A30713). Treat HPA’s Approved rating as provisional because its staining and RNA consistency is medium and external verification is pending (HPA: tissue IHC reliability).
Boster reagents
Best GTPBP2 / GTP-binding protein 2 IHC Antibodies
Anti-GTPBP2 A30713 is listed for IHC, IF, and ICC in human and mouse (catalog applications/reactivity). Its supplied figure documents paraffin-section IHC in human colon carcinoma (catalog IHC image caption).
Real IHC data
Anti-GTPBP2 Antibody
Cat # A30713
A30713 is listed for IHC, IF, ICC, and ELISA in human and mouse (catalog applications/reactivity). Its IHC image shows paraffin-embedded human colon carcinoma with a peptide-blocked comparison (catalog IHC image caption); no IF image is supplied (catalog IF images).
Which to pick: Choose A30713 for tissue IHC on paraffin sections: its own image shows human colon carcinoma staining with a peptide-blocked comparison (catalog IHC image caption), and its listed IHC dilution is 1:100–1:300 (catalog IHC dilution). For IF/ICC, A30713 lists both applications and an IF dilution of 1:50 (catalog applications/IF dilution); no IF image is supplied (catalog IF images). For work involving human and mouse samples, A30713 is a rabbit polyclonal antibody with listed reactivity to both species (catalog host, clonality, and reactivity); the fixative used for its paraffin-section image is unreported (catalog IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BX10 (GTPB2_HUMAN, GTP-binding protein 2).
- Human Protein Atlas. GTPBP2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. GTPBP2 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. GTPBP2 antibody validation summary (2 antibodies).
- GTPBP2 positively regulates the invasion, migration and proliferation of non-small cell lung cancer. Journal of Cancer 2021 — PMC8176249.
- GTPBP2 inactivates Hippo signaling to promote triple-negative breast cancer cell malignancy. Scientific reports 2026 — PMC13057012.
- GTP binding protein 2 maintains the quiescence, self-renewal, and chemoresistance of mouse colorectal cancer stem cells via promoting Wnt signaling activation. Heliyon 2024 — PMC10926081.
- GTPBP1 resolves paused ribosomes to maintain neuronal homeostasis. eLife 2020 — PMC7665888.
- PubMed PMID:11054535 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GTPBP2 (GTP-binding protein 2). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A30713 at 1:100 - 1:300 (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 colon carcinoma tissue; fixative not specified (datasheet A30713); 2. Fixation: Image fixative and duration unreported (datasheet A30713); 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-GTPBP2, 1:100 - 1:300 (datasheet A30713); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Spleen red pulp lacks staining despite reported spleen expression (HPA tissue IHC; UniProt); Isoform / epitope: 4 isoforms; epitope coverage is unresolved (UniProt; datasheet); Regulation: Tissue-dependent expression; no regulator specified (UniProt). Evidence: UniProt Q9BX10, HPA tissue IHC · Approved, HPA ICC-IF · vesicles., Datasheet A30713, PMC8176249, PMC10926081. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GTPBP2/immunohistochemistry-ihc-guide.