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GRB10 / Growth factor receptor-bound protein 10 · IHC design guide
Design Immunohistochemistry for GRB10
Plan GRB10 staining in paraffin sections around the cytoplasmic pattern seen in most tissues (HPA tissue IHC). Use colon glandular cells as a positive reference (HPA tissue IHC), and optimize the IHC-validated antibody within 2–5 μg/mL (datasheet A01663-2).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GRB10 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01663-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue |
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 | Adipocytes may lack detectable staining (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 4 isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended GRB10 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet A01663-2). Three published GRB10 IHC protocols provide tissue-specific comparisons (PMC11367874; PMC10449302; PMC4795681).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A01663-2) |
| Fixation | Image fixative and duration unreported (datasheet A01663-2); 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 A01663-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01663-2) |
| Primary antibody | Rabbit anti-GRB10, 2-5μg/ml (datasheet A01663-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01663-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01663-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GRB10-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet A01663-2); the mouse kidney protocol used microwave citrate retrieval at pH 6.0–6.3 (PMC4795681 methods).
Section 2
What Is the Expected GRB10 Staining Pattern?
GRB10 is a cytoplasmic adapter protein with no transmembrane segment (UniProt Q13322: subcellular location and topology). In paraffin section IHC, expect cytoplasmic staining in many tissues, with strong signal reported in selected glandular cells, cardiomyocytes and neurons (HPA: tissue IHC). HPA rates the tissue pattern Enhanced, while describing medium consistency between antibody staining and RNA expression (HPA: reliability).
| Cytoplasmic staining in appendix or colon glandular cells, with tissue structure preserved. | This matches the reported compartment and strongly staining cell types (HPA: cytoplasmic expression in most tissues; High in appendix and colon glandular cells). Judge intensity within the relevant cells, since neighboring cells can differ. |
| Strong signal confined mainly to nuclei, with little cytoplasmic signal. | A predominantly nuclear pattern conflicts with the reported cytoplasmic IHC pattern (HPA: tissue IHC) and cytoplasmic localization (UniProt Q13322: subcellular location). Check the counterstain and detection controls before assigning it to GRB10 (general IHC practice). |
| Strong adipocyte staining, especially when the expected positive cells are faint. | Adipocytes were reported as Not detected (HPA: adipose tissue IHC). Investigate antibody cross-reactivity or endogenous detection activity (general IHC practice); this observation alone cannot identify which caused the signal. |
| Diffuse color across stroma, lumens and cells, obscuring cell boundaries. | This is difficult to score as cell-specific cytoplasmic staining (HPA: expected cytoplasmic pattern). Review blocking, antibody concentration and wash conditions as general sources of IHC background (general IHC practice). |
| No signal in a known positive reference section processed alongside the test section. | For example, HPA reports High staining in Purkinje cells and cardiomyocytes (HPA: cerebellum and heart muscle IHC). A blank positive reference calls for checking antibody, retrieval and detection performance before interpreting a blank test section (general IHC practice). |
💡Expected GRB10 appearanceCall a positive result when defined cells show predominantly cytoplasmic chromogenic signal, potentially strong in HPA High cell types; diffuse tissue-wide deposits or nuclear-only staining warrant investigation (HPA: tissue IHC; UniProt Q13322: cytoplasm; general IHC practice).
| Compartment and cell selection | Use the cell type and compartment together when scoring: HPA reports cytoplasmic expression in most tissues, but High staining specifically in listed glandular, cardiac and neuronal cells (HPA: tissue IHC). |
| Reference tissue choice | Appendix or colon glandular cells offer reported High-staining examples; adipocytes are reported Not detected (HPA: tissue IHC). Treat these as observed reference patterns, not guarantees for every specimen or antibody. |
| Antibody validation | HPA027502 is IHC Enhanced and CAB019423 is IHC Approved; HPA031818 has no stated IHC status (HPA: antibody validation). Match the validation record to the antibody actually used. |
| Isoforms and epitope | UniProt lists four GRB10 isoforms and a full-length chain spanning residues 1–594 (UniProt Q13322: isoforms and processing). The supplied record gives no antibody epitope, so isoform coverage cannot be inferred. |
| Antigen retrieval | Select and optimize retrieval against a positive reference and background controls (general IHC practice). Target-specific fixation sensitivity and an optimal GRB10 retrieval condition are unreported in the supplied UniProt and HPA evidence. |
| IF/ICC comparison | Q: Should punctate IF/ICC signal match paraffin section IHC? A: HPA supports a mainly vesicular ICC-IF location, while tissue IHC is described as cytoplasmic (HPA: subcellular and tissue IHC). Interpret each application in its own context. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive reference and test section are both blank. | Antibody, retrieval or detection may have failed (general IHC practice). | Check the reference tissue, reagent steps and chromogen development together; HPA reports High staining in appendix glandular cells, among other examples (HPA: tissue IHC). |
| Reference stains, but the test section is blank. | The sampled cell type may have little detectable signal; HPA reports Not detected in adipocytes and Low in oral mucosal squamous cells (HPA: tissue IHC). | Identify and score the relevant cells first. Report a negative result for the sampled cells without assuming that GRB10 is absent from every tissue compartment (general IHC practice). |
| The entire section has weak, diffuse color. | Excess antibody, insufficient washing or nonspecific detection can raise background (general IHC practice). | Review antibody dilution, blocking and washes; compare a reagent control before calling weak cytoplasmic signal positive (general IHC practice). |
| Dark deposits appear in unrelated cells or tissue spaces. | Endogenous enzyme activity or precipitated chromogen may mimic specific staining (general IHC practice). | Inspect a detection control and the deposit shape; apply an appropriate endogenous activity block when indicated by the detection chemistry (general IHC practice). |
| The apparent signal is mainly nuclear. | The observed compartment differs from the expected cytoplasmic pattern (HPA: tissue IHC; UniProt Q13322: subcellular location). | Compare with the counterstain and a known positive section; resolve staining controls before assigning the nuclear pattern to GRB10 (general IHC practice). |
| Strong staining appears in adipocytes. | This conflicts with HPA's Not detected adipocyte observation, although a single image does not establish cross-reactivity (HPA: adipose tissue IHC). | Compare an antibody control and an HPA High cell type stained in the same run; assess whether the adipocyte signal persists independently (general IHC practice). |
Experiment evidence
IHC & IF images showing GRB10 staining
A01663-2 · IHCIHC analysis of GRB10 using anti-GRB10 antibody (A01663-2). GRB10 was detected in a paraffin-embedded section of human colorectal adenocarci
A01663-2 · IHCIHC analysis of GRB10 using anti-GRB10 antibody (A01663-2). GRB10 was detected in a paraffin-embedded section of human laryngeal squamous ce
A01663 · IFImmunofluorescence of GRB10 in rat brain cells with GRB10 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000106070-GRB10/tissue/cerebral+cortex#imid_5014804
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000106070-GRB10/tissue/colon#imid_5014730
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000106070-GRB10/tissue/liver#imid_5014731
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000106070-GRB10/tissue/kidney#imid_5014724
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA031818; green = GRB10, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA031818; green = GRB10, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GRB10 IHC & IF
🧪Run colon first: its glandular cells should stain (HPA: High in colon glandular cells). Run adipose tissue as the negative, focusing on adipocytes (HPA: Not detected in adipocytes); on the colon slide, assess adjacent nonglandular cells as internal background only if they show little or no staining, since their GRB10 status is not specified by the supplied HPA rows.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GRB10 in A-549, U-251MG, U2OS, with annotated localisation: Vesicles (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section; a concentration-matched rabbit IgG control appropriate to the primary antibody’s clonality; and GRB10 knockout material or a peptide-blocked primary if the immunizing peptide is available (standard IHC control practice; caption: rabbit anti-GRB10 primary). On colon sections, quench endogenous peroxidase and check luminal debris and stroma for nonspecific DAB signal (standard chromogenic IHC practice).
⚠️Feasibility: The selected A01663-2 paraffin-section caption uses heat retrieval in EDTA at pH 8.0 and a primary concentration of 2 μg/ml, but does not report a fixative or a target-specific fixation window or effect; the need for retrieval has not been established (selected-SKU tissue-IHC caption). Frozen-section performance is unreported; IF/ICC has supported vesicular localization in A-549, U-251MG and U2OS images, but the supplied evidence does not establish that either approach is easier than paraffin IHC (HPA subcellular). In colon, distinguish glandular staining from nonspecific luminal or stromal DAB signal (HPA: High in colon glandular cells; standard chromogenic IHC practice).
HPA tissue IHC evidence for GRB10
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GRB10 IHC Tips
Troubleshoot GRB10 staining in paraffin sections by checking retrieval, cellular localisation, tissue controls and scoring before interpreting chromogenic signal.
Which retrieval condition should I try first for weak GRB10 staining?
Start with heat-mediated retrieval in EDTA, pH 8.0 for paraffin sections (datasheet A01663-2). The selected tissue image used this condition before overnight incubation with 2 μg/ml antibody at 4°C (datasheet A01663-2). If staining is weak, compare carefully timed heating and cooling conditions on adjacent sections while keeping antibody concentration, detection and exposure to DAB constant (standard IHC practice). Include a positive-control section with an expected cytoplasmic signal, such as colon glandular cells (HPA: High in colon glandular cells). Judge retrieval by improved cellular staining without a parallel rise in diffuse background or tissue damage (standard IHC practice).
Could fixation explain weak or uneven GRB10 staining?
The selected paraffin-section caption does not report its fixative, so GRB10-specific sensitivity to fixation is unknown (datasheet A01663-2). Record the fixative, fixation duration and processing history for each specimen, then compare sections processed alike when troubleshooting weak staining (standard IHC practice). Uneven fixation can complicate interpretation of chromogenic tissue staining, but these payloads do not establish a GRB10-specific fixation effect (standard IHC practice; datasheet A01663-2). Apply the documented EDTA, pH 8.0 retrieval condition consistently before changing other variables (datasheet A01663-2). Use expected cytoplasmic staining in colon glandular cells as a tissue-level reference, without treating its intensity as a measure of fixation quality (HPA: High in colon glandular cells; HPA: cytoplasmic expression in most tissues).
Should GRB10 stain the cytoplasm uniformly or appear punctate?
Assess chromogenic GRB10 primarily in the cytoplasmic compartment: tissue IHC reports cytoplasmic expression in most tissues (HPA: tissue IHC profile). A punctate component is plausible because the subcellular dataset places GRB10 mainly in vesicles (HPA: vesicles, supported). GRB10 can accompany internalising IGF1R into early endosomes, although that annotation is by similarity and does not establish the identity of puncta in a section (UniProt Q13322: subcellular location). It has no transmembrane segment, so a crisp, continuous cell-surface rim warrants scrutiny against controls (UniProt Q13322: topology). Compare glandular-cell staining in colon with adjacent background and assess nuclear-only deposits cautiously (HPA: High in colon glandular cells; HPA: tissue IHC profile).
How could isoforms or epitope placement affect GRB10 IHC?
GRB10 has four listed isoforms, so check the antibody’s documented immunogen or epitope before assuming it detects every variant (UniProt Q13322: isoforms). Its Ras-associating, PH and SH2 regions span residues 166–250, 290–399 and 493–574, respectively (UniProt Q13322: domains). The record also lists phosphorylation at several sites, including serines 150 and 428; the payload does not establish whether those modifications alter this antibody’s staining (UniProt Q13322: modified residues). Test retrieval and antibody concentration on matched serial sections before attributing a missing signal to an isoform or modification (standard IHC practice). Report the epitope’s documented position if available, alongside the scoring method (standard IHC practice).
How can IF help investigate a puzzling chromogenic GRB10 pattern?
Use IF as a separate corroborating experiment and multiplex GRB10 with a marker for the cell type being assessed in IHC, such as glandular cells in colon (HPA: High in colon glandular cells; standard IF practice). Choose a fluorophore channel with low tissue autofluorescence and inspect unstained tissue plus single-label controls before interpreting overlap (standard IF practice). Because GRB10 lacks a transmembrane segment and is reported in cytoplasmic vesicles, use permeabilisation appropriate for access to intracellular epitopes (UniProt Q13322: topology; HPA: vesicles, supported; standard IF practice). The IF dataset includes images from A-549, U-251MG and U2OS, but their fixation conditions are not supplied here (HPA: subcellular dataset). Do not transfer the paraffin-section retrieval result into an IF fixation claim (datasheet A01663-2).
What should I check when GRB10 DAB staining is diffuse?
The selected paraffin-section workflow used 10% goat serum, an anti-rabbit peroxidase-conjugated secondary and DAB detection (datasheet A01663-2). Check the no-primary control, apply a peroxidase block and control DAB development time when diffuse brown signal obscures cellular boundaries (standard chromogenic IHC practice). Compare background in tissue outside the expected cell population with cytoplasmic signal in a positive reference, such as colon glandular cells (HPA: High in colon glandular cells; HPA: tissue IHC profile). Titrate the primary around the documented 2 μg/ml condition while holding retrieval and detection constant (datasheet A01663-2; standard IHC practice). Treat widespread extracellular deposits as suspect until they clear in controls (standard IHC practice).
How should I quantify GRB10 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, because tissue IHC describes GRB10 as cytoplasmic in most tissues (HPA: tissue IHC profile). For comparable sections, report the percentage of positive cells and an H-score based on the proportion at each staining intensity; keep thresholds fixed across specimens (standard IHC practice). If counting discrete positive cells, report density per mm² of viable tissue and exclude empty space or necrosis (standard IHC practice). Normalise cell counts to the area or number of evaluable cells within the same defined compartment, and include matched negative controls (standard IHC practice). Record whether puncta contribute to the cytoplasmic score, since vesicular localisation is supported (HPA: vesicles, supported).
How can I distinguish credible GRB10 positivity from staining artefact?
Credible tissue signal should map to identifiable cytoplasm, with possible puncta, and remain distinguishable from the no-primary control (HPA: tissue IHC profile; HPA: vesicles, supported; standard IHC practice). Colon glandular cells offer a reported high-expression reference, whereas adipocytes are reported as not detected in adipose tissue (HPA: High in colon glandular cells; HPA: adipocytes not detected). Treat isolated nuclear staining, section-edge enhancement, necrotic deposits and persistent brown signal after omitting primary antibody as reasons to investigate artefact (standard IHC practice; HPA: tissue IHC profile). Repeat suspicious areas on adjacent sections with consistent EDTA, pH 8.0 retrieval and a peroxidase block (datasheet A01663-2; standard IHC practice). HPA reports medium agreement between antibody staining and RNA expression, so avoid equating staining intensity with transcript abundance (HPA: Enhanced reliability description).
Boster reagents
Best GRB10 / Growth factor receptor-bound protein 10 IHC Antibodies
Anti-GRB10 antibodies have IHC images from paraffin sections of human cancers and mouse brain (catalog: A01663-2 IHC captions), plus an IF image from rat brain cells (catalog: A01663 IF caption).
Real IHC data
Anti-GRB10 Antibody ®
Cat # A01663-2
Real IF data
Anti-GRB10 Antibody
Cat # A01663
A01663-2 has IHC images from paraffin sections of human colorectal adenocarcinoma, laryngeal squamous cell carcinoma, liver cancer, and mouse brain (catalog: A01663-2 IHC captions). A01663 has an IF image from rat brain cells at 20 μg/mL (catalog: A01663 IF caption).
Which to pick: Choose A01663-2 for chromogenic tissue IHC: its caption documents paraffin sections, EDTA retrieval at pH 8.0, and DAB detection; the fixative is unreported (catalog: A01663-2 IHC caption). Choose A01663 for cell IF/ICC work, starting from its rat brain cell IF example at 20 μg/mL; ICC is not separately listed as an application (catalog: A01663 applications and IF caption). For cross-species planning, A01663-2 lists human, monkey, mouse, and rat reactivity, but its IHC images document human and mouse samples (catalog: A01663-2 reactivity and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13322 (GRB10_HUMAN, Growth factor receptor-bound protein 10).
- Human Protein Atlas. GRB10 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GRB10 subcellular location (ICC-IF): Mainly localized to vesicles..
- Human Protein Atlas. GRB10 antibody validation summary (3 antibodies).
- Combination of arsenic trioxide and apatinib synergistically inhibits small cell lung cancer by down-regulating VEGFR2/mTOR and Akt/c-Myc signaling pathway via GRB10. Hereditas 2024 — PMC11367874.
- GRB10 is a novel factor associated with gastric cancer proliferation and prognosis. Aging 2023 — PMC10449302.
- GRB10 as a pan-cancer biomarker linking oncogenic signaling, immunosuppression, and immunotherapy resistance. Discover oncology 2025 — PMC12748335.
- Amelioration of Diabetic Mouse Nephropathy by Catalpol Correlates with Down-Regulation of Grb10 Expression and Activation of Insulin-Like Growth Factor 1 / Insulin-Like Growth Factor 1 Receptor Signaling. PloS one 2016 — PMC4795681.
- PubMed PMID:7479769 — UniProt-cited evidence.
- PubMed PMID:8798417 — UniProt-cited evidence.
- PubMed PMID:9006901 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GRB10 (Growth factor receptor-bound protein 10). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01663-2). Validated IHC antibody: A01663-2 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A01663-2); 2. Fixation: Image fixative and duration unreported (datasheet A01663-2); 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 A01663-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01663-2); 8. Primary antibody: Rabbit anti-GRB10, 2-5μg/ml (datasheet A01663-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01663-2). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Adipocytes may lack detectable staining (HPA tissue IHC); Isoform / epitope: 4 isoforms; check antibody epitope coverage (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q13322, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to vesicles., Datasheet A01663-2, PMC11367874, PMC10449302. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GRB10/immunohistochemistry-ihc-guide.