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GRB10 / Growth factor receptor-bound protein 10 · IHC design guide
Design Immunohistochemistry for GRB10
Plan GRB10 chromogenic IHC-P using glandular cells and cardiomyocytes as strongly staining examples (HPA tissue IHC). Start the catalog antibody at 2–5 μg/mL (datasheet A01663-2) and compare with adipocytes, where staining was not detected (HPA tissue IHC).
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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across 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 sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation is unspecified (UniProt) | |
| Isoform / epitope | 4 isoforms; check epitope coverage (UniProt) |
Section 1
Recommended GRB10 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 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). For mouse kidney, a published protocol used citrate pH 6.0–6.3 (PMC4795681).
Section 2
What Is the Expected GRB10 Staining Pattern?
In paraffin-section IHC, expect GRB10 mainly in the cytoplasm of many cell types, including strongly stained glandular cells, cardiomyocytes and neurons (HPA: tissue IHC). GRB10 has no transmembrane segment (UniProt Q13322 topology). HPA rates its IHC evidence Enhanced, while reporting medium consistency between staining and RNA expression (HPA: tissue IHC).
| Cytoplasmic staining in glandular cells of appendix, colon, duodenum, endometrium or gallbladder. | This matches the reported high staining in these cells (HPA: tissue IHC). Judge the named cells and their cytoplasm, rather than treating every stained structure in the section as equally informative. |
| Strong cytoplasmic staining in cardiomyocytes, cerebellar Purkinje cells or hippocampal neurons. | These are further reported high-staining cell populations (HPA: tissue IHC). Their presence can help assess a run, although HPA reports only medium consistency between antibody staining and RNA expression (HPA: tissue IHC). |
| Predominantly nuclear or sharply membrane-rimmed staining. | That distribution conflicts with the mainly cytoplasmic tissue profile (HPA: tissue IHC) and the absence of a transmembrane segment (UniProt Q13322 topology). Check whether the pattern persists after detection controls before assigning it to GRB10. |
| Strong adipocyte staining, especially when the expected glandular or neuronal cells are unstained. | HPA reports GRB10 as not detected in adipocytes (HPA: adipose tissue IHC). Consider cross-reactivity or endogenous detection activity; a single unexpected cell type does not identify which mechanism caused the signal. |
| Broad, fairly even chromogen over cells and surrounding tissue, or no signal in an HPA high-staining cell population. | Neither result demonstrates the expected cell-associated pattern (HPA: tissue IHC). Background calls for detection controls; absent signal calls for checks of tissue, antibody and staining workflow before a biological conclusion. |
💡Expected GRB10 appearanceCall a section positive when cytoplasmic staining is clear in an HPA high-staining cell population, such as colon glandular cells or cardiomyocytes (HPA: tissue IHC); predominant nuclear staining or strong adipocyte staining is discordant (HPA: tissue IHC).
| Cell and tissue choice | HPA reports high staining in several glandular, cardiac and neuronal populations, but low staining in oral and vaginal squamous cells and bone marrow hematopoietic cells (HPA: tissue IHC). Use a reported high population to assess detection. |
| Subcellular interpretation | GRB10 is annotated as cytoplasmic and lacks a transmembrane segment (UniProt Q13322). Its association with early endosomes during IGF1R internalization is described by similarity (UniProt Q13322), so puncta alone are not an IHC identity test. |
| Antibody evidence | IHC status is Enhanced for HPA027502 and Approved for CAB019423; HPA031818 has no listed IHC status (HPA: antibody validation). These statuses describe different evidence for the antibodies, not a guarantee for every section. |
| Isoforms and modified residues | UniProt lists four isoforms and multiple phosphorylated residues (UniProt Q13322). The payload gives no antibody epitope, so it cannot establish isoform coverage or a phosphorylation-dependent IHC pattern. |
| IF/ICC Q: Should vesicular fluorescence be expected? | A: HPA reports a supported main location in vesicles, with ICC-IF images from A-549, U-251MG and U2OS (HPA: subcellular). Interpret that observation in the separate IF/ICC guide; it supplies no IHC retrieval or dilution setting. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in a reported high-staining population. | A failed detection step or unsuitable run conditions are possible; high staining is reported for colon glandular cells, among others (HPA: tissue IHC). | Check that the named cells are present, then review the antibody's IHC instructions, retrieval and detection controls. No GRB10-specific retrieval condition is supplied. |
| Only weak stain in oral or vaginal squamous cells. | Low staining is reported in those cells (HPA: tissue IHC), so this result alone gives a limited test of assay sensitivity. | Assess a reported high-staining population, such as appendix glandular cells (HPA: tissue IHC), within the same staining run where feasible. |
| Strong stain appears in adipocytes. | This conflicts with HPA's not-detected adipocyte result (HPA: adipose tissue IHC); cross-reactivity or endogenous detection activity is possible. | Inspect a no-primary control and compare cell boundaries and surrounding tissue. Treat the adipocyte signal as unresolved until the source is identified. |
| Nuclei dominate the stain. | A nuclear-dominant result disagrees with the cytoplasmic tissue profile (HPA: tissue IHC; UniProt Q13322 subcellular location). | Compare with a reported high-staining cell population and a no-primary control. Recheck scoring so nuclear counterstain is not mistaken for chromogen. |
| Chromogen coats many structures without clear cell boundaries. | Diffuse background can obscure the cell-associated cytoplasmic pattern (HPA: tissue IHC); the payload does not identify a GRB10-specific cause. | Inspect the no-primary control and review blocking, washes and detection steps as general IHC practice. Avoid scoring diffuse deposition as positive cells. |
| Two antibodies give different cell patterns. | HPA lists differing IHC validation statuses and reports medium staining-to-RNA consistency (HPA: antibody validation; tissue IHC). Discordance needs investigation. | Compare the same named cell populations and controls for each antibody. Report the disagreement instead of assigning an unsupported GRB10-specific pattern. |
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: glandular cells should show strong GRB10 staining (HPA: High in colon glandular cells). Use adipose tissue as the negative tissue, where adipocytes are not detected (HPA: Not detected in adipocytes); on the colon slide, nonglandular cells should show only background relative to the glands, but their GRB10 status must be verified rather than assumed (HPA: only glandular cells are specified for colon).
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 control and a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality (IHC caption: rabbit primary antibody). A GRB10 knockout sample provides a biological negative control; quench endogenous peroxidase in colon sections before HRP–DAB detection (IHC caption: colon-derived section with HRP–DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01663-2 tissue-IHC caption does not state the fixative (IHC caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (IHC caption: EDTA retrieval); the supplied evidence does not establish whether frozen sections or IF are easier. Colon sections may show endogenous peroxidase background with HRP–DAB detection, so interpret staining against the quenched controls (IHC caption: HRP–DAB detection; standard 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, cytoplasmic localisation, cell type and controls against the available tissue evidence.
What retrieval should I try first when GRB10 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01663-2). The selected tissue image used that retrieval before an overnight incubation at 4°C with 2 μg/ml primary antibody (caption A01663-2). If staining remains weak, compare a longer or shorter heating period on adjacent sections while keeping the antibody and detection conditions fixed (standard IHC practice). Check that improved signal remains within cells rather than appearing uniformly across damaged tissue (standard IHC practice). The caption reports paraffin embedding but does not identify the fixative, so it cannot establish a fixation-specific retrieval adjustment (caption A01663-2).
Could fixation explain weak or uneven GRB10 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (caption A01663-2). Record the fixative and fixation duration for each specimen, then compare adjacent sections processed with the same pH 8.0 EDTA retrieval and 2 μg/ml primary concentration used in the image (caption A01663-2; standard IHC practice). Review morphology and staining together, because poorly preserved or unevenly processed regions can make an apparent expression difference unreliable (standard IHC practice). Keep detection and counterstaining matched across the comparison (standard IHC practice). Do not infer a GRB10-specific fixation effect from its cytoplasmic annotation or tissue staining pattern (UniProt Q13322; HPA tissue IHC).
Where should GRB10 staining appear in a convincing section?
Expect predominantly cytoplasmic staining in tissue sections (HPA tissue IHC: cytoplasmic expression in most tissues; UniProt Q13322: cytoplasm). GRB10 has no transmembrane segment, so an exclusively crisp cell-surface rim warrants scrutiny rather than automatic assignment to the target (UniProt Q13322 topology). Vesicular staining is biologically plausible: cell imaging places GRB10 mainly in vesicles, and the protein can accompany internalised IGF1R to early endosomes (HPA subcellular; UniProt Q13322, by similarity). Compare the candidate pattern with neighbouring cells and a no-primary control under the same pH 8.0 retrieval conditions (datasheet A01663-2; standard IHC practice). Interpret granular DAB cautiously when tissue pigment or precipitate is present (standard IHC practice).
Can isoforms or epitope location change how I interpret GRB10 staining?
GRB10 has 4 annotated isoforms, while the supplied caption does not identify the antibody epitope (UniProt Q13322; caption A01663-2). Therefore, do not assume that this antibody detects every isoform or distinguishes one isoform from another (UniProt Q13322 isoforms; caption A01663-2). If epitope documentation becomes available, map it against the Ras-associating region at residues 166–250, PH domain at 290–399, and SH2 domain at 493–574 (UniProt Q13322). GRB10 also has annotated phosphorylation sites, including serines 150 and 428; their effect on this antibody's staining is unreported (UniProt Q13322; caption A01663-2). Compare staining with an independently validated reagent before assigning an isoform-specific explanation (standard IHC practice).
How can I assess GRB10 by multiplex immunofluorescence?
Treat this as a separate assay: the selected antibody's supplied tissue example uses chromogenic detection, with no IF conditions reported (caption A01663-2). Pair GRB10 with a marker for the expected cell type, such as a glandular epithelial marker when examining colon, where glandular cells stain highly (HPA tissue IHC: colon glandular cells, High; standard IF practice). Choose spectrally separated fluorophores and consider a far-red channel when tissue autofluorescence interferes with shorter wavelengths (standard IF practice). Because GRB10 is cytoplasmic and has no transmembrane segment, optimise permeabilisation to expose intracellular epitopes while preserving morphology (UniProt Q13322 topology; standard IF practice). Include single-stain and no-primary controls to assess bleed-through and background (standard IF practice).
How do I separate GRB10 signal from chromogenic background?
Begin with a no-primary section and inspect whether DAB deposits persist in the same structures (standard IHC practice). The selected example used a 10% goat-serum block, peroxidase-conjugated secondary antibody and DAB development, but it does not report a dedicated endogenous-peroxidase blocking step (caption A01663-2). Add and verify a peroxidase block as a general chromogenic IHC control, especially where the no-primary section develops colour (standard IHC practice). Check that the secondary reagent matches the rabbit primary and that washes remove loosely bound reagent before DAB development (caption A01663-2; standard IHC practice). Assess tissue edges, folds and precipitate separately from intracellular staining (standard IHC practice).
How should I score GRB10 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable tissue region before scoring, then record the percentage of GRB10-positive cells and staining intensity separately (standard IHC practice). For an H-score, sum the percentage at each intensity multiplied by its 0–3 grade, yielding a 0–300 scale (standard IHC practice). Normalise cell counts to the number of evaluable cells, or report positive-cell density per mm² of viable tissue when cell counting is impractical (standard IHC practice). In colon, score glandular cells consistently because they are a documented high-staining population (HPA tissue IHC: colon glandular cells, High). Hold retrieval, DAB development and counterstaining constant across compared sections (datasheet A01663-2; standard IHC practice).
When should a GRB10-positive result be considered credible?
A credible result has cellular, predominantly cytoplasmic staining in an appropriate population, consistent with the tissue profile and localisation annotations (HPA tissue IHC; UniProt Q13322). Colon glandular cells are a documented high-staining population, whereas adipocytes in adipose tissue were not detected in the cited tissue survey (HPA tissue IHC). Reject isolated staining confined to section edges, folds or necrotic regions as insufficient evidence of GRB10 expression (standard IHC practice). Compare the pattern with a no-primary control to identify endogenous enzyme activity, pigment or DAB precipitate (standard IHC practice). The tissue atlas reports Enhanced reliability with medium staining-to-RNA consistency, so resolve discordant findings with independent evidence (HPA tissue IHC).
Boster reagents
Best GRB10 / Growth factor receptor-bound protein 10 IHC Antibodies
Anti-GRB10 antibodies have real IHC data from human tumor and mouse brain paraffin sections (A01663-2 image captions), plus IF data from rat brain cells (A01663 image caption).
Real IHC data
Anti-GRB10 Antibody ®
Cat # A01663-2
Real IF data
Anti-GRB10 Antibody
Cat # A01663
A01663-2 lists IHC and shows staining in human colorectal adenocarcinoma, laryngeal squamous cell carcinoma, and liver cancer paraffin sections, plus mouse brain paraffin sections (catalog applications; A01663-2 image captions). A01663 lists IF and shows staining in rat brain cells (catalog applications; A01663 image caption).
Which to pick: Choose A01663-2 for tissue IHC: its paraffin section images use heat retrieval in EDTA at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (A01663-2 image captions). Choose A01663 for IF in cells, starting at 20 μg/mL based on its rat brain cell image; no ICC result is reported (A01663 image caption; catalog applications). For cross-species selection, A01663-2 lists human, monkey, mouse, and rat reactivity, while A01663 lists human, mouse, and rat; clonality is unreported for both (catalog reactivity; catalog clone fields).
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 in most tissues (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 sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 4 isoforms; check epitope coverage (UniProt); Regulation: Expression regulation is unspecified (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.