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ARHGEF7 / Rho guanine nucleotide exchange factor 7 · IHC design guide
Design Immunohistochemistry for ARHGEF7
Use the IHC-validated antibody on paraffin sections (datasheet A02764-2) and assess cytoplasmic staining (HPA tissue IHC). Compare a medium-staining tissue with a reported low or undetected tissue when optimizing signal (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ARHGEF7 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 signal (HPA tissue IHC); adhesion-associated protein (UniProt) | |
| Staining pattern | Broad cytoplasmic staining; cell-type intensity varies (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02764-2) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Salivary gland+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A02764-2); verify before use. | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation not established (UniProt) | |
| Isoform / epitope | 6 isoforms; epitope coverage needs checking (UniProt) |
Section 1
Recommended ARHGEF7 IHC & IF Protocols
The catalog antibody's IHC-P protocol (datasheet A02764-2) is accompanied by a published colorectal tissue protocol (PMC6192735).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A02764-2) |
| Fixation | Image fixative and duration unreported (datasheet A02764-2); 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 A02764-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02764-2) |
| Primary antibody | Rabbit anti-ARHGEF7, 2-5 μg/ml (datasheet A02764-2) |
| Primary incubation | Overnight at 4 °C (datasheet A02764-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02764-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ARHGEF7-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A02764-2); the published protocol specifies 1 mM EDTA at pH 8.0 (PMC6192735).
Section 2
What Is the Expected ARHGEF7 Staining Pattern?
ARHGEF7 is a cytoplasmic protein found at the cell cortex, ruffles, lamellipodia and cell junctions; a small proportion occurs at focal adhesions (UniProt Q14155). Expect cytoplasmic staining in multiple cell types, including adipocytes, glandular cells and cerebral cortical neurons (HPA tissue IHC). HPA calls the tissue IHC profile Approved, with medium consistency between staining and RNA expression (HPA tissue IHC). ARHGEF7 has no transmembrane segment (UniProt Q14155 topology).
| Cytoplasmic staining in adipocytes, glandular cells or cerebral cortical neurons. | This fits the reported ubiquitous cytoplasmic profile. HPA records medium staining in adipocytes, several glandular populations and cortical neurons; compare intensity within the same cell population (HPA tissue IHC). |
| Cortical or junction associated accentuation accompanies cytoplasmic staining. | This can fit ARHGEF7 localisation at the cell cortex, cell junctions, ruffles and lamellipodia (UniProt Q14155). Focal adhesion staining, if resolvable, should involve only a small proportion of the protein (UniProt Q14155). |
| Predominantly nuclear staining, with little cytoplasmic signal. | The dominant compartment conflicts with the HPA cytoplasmic profile and UniProt localisation (HPA tissue IHC; UniProt Q14155). Treat it as a possible artefact and check the controls before scoring it as ARHGEF7. |
| Strong staining in salivary glandular cells or smooth muscle cells. | HPA reports ARHGEF7 as not detected in those cell populations (HPA tissue IHC). Unexpected staining raises concern for cross-reactivity or endogenous detection activity; it does not establish either cause without control sections. |
| Diffuse haze throughout the section, or no signal in a reported positive cell population. | Neither result cleanly supports the expected cell specific cytoplasmic pattern (HPA tissue IHC). Diffuse haze calls for background controls; absent signal in a reported medium staining population calls for a check of tissue and detection controls (standard IHC practice). |
💡Expected ARHGEF7 appearanceCall positive when cytoplasmic staining is discernible in HPA medium staining cells, such as adipocytes or cortical neurons; a chiefly nuclear pattern or strong staining in HPA not detected cells warrants investigation (HPA tissue IHC; UniProt Q14155).
| Tissue and cell population | HPA reports medium staining in adipocytes, adrenal and other listed glandular cells, glia in caudate, cerebellar granular layer cells and cortical neurons; thyroid and colon glandular cells are low (HPA tissue IHC). Match the comparison to the named cell population. |
| Compartment and topology | ARHGEF7 is cytoplasmic, with cortical and adhesion associated localisation, and has no transmembrane segment (UniProt Q14155). A membrane edge accent may be plausible, but the record does not predict a uniform membrane outline. |
| Antibody evidence | HPA lists HPA004744 as Approved for IHC, while the tissue profile has medium consistency with RNA data (HPA antibodies; HPA tissue IHC). Interpret unexpected patterns with controls; Approved is not an Enhanced validation claim. |
| Isoforms and processing | UniProt lists six isoforms and a chain spanning residues 1–803, with no signal peptide or propeptide (UniProt Q14155). The supplied record gives no antibody epitope, so it cannot predict which isoforms this antibody detects. |
| Can IF/ICC confirm this IHC pattern? | HPA provides no main ICC/IF location or cell line images, and its listed antibody has no ICC status (HPA subcellular; HPA antibodies). UniProt localisation is useful context, but it does not supply an IF validation result (UniProt Q14155). |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported medium staining cells show no chromogenic signal. | The run may have failed, or the examined section may lack the named positive cell population (HPA tissue IHC; standard IHC practice). | Confirm the cell population on the counterstained section, then check the run's positive tissue and detection controls (standard IHC practice). Do not infer target specific fixation sensitivity; none is reported here. |
| Nuclear staining dominates the slide. | This differs from HPA's cytoplasmic profile and UniProt's cortical and adhesion associated localisation (HPA tissue IHC; UniProt Q14155). | Review morphology and no primary control staining, then reassess antibody specificity before assigning the nuclear signal to ARHGEF7 (standard IHC practice). |
| Salivary glandular or smooth muscle cells stain strongly. | HPA records both populations as not detected; cross-reactivity or endogenous detection activity is possible, but the image alone cannot distinguish them (HPA tissue IHC; standard IHC practice). | Compare a no primary control and the expected positive cell populations in the same run; investigate any persistent cell specific signal (standard IHC practice). |
| Diffuse chromogen obscures cell borders. | Widespread background prevents a reliable comparison with HPA's cytoplasmic cell staining (HPA tissue IHC). Detection background is a possible general IHC cause (standard IHC practice). | Check no primary and reagent controls, washing and detection conditions; score ARHGEF7 only where cellular boundaries and cytoplasm remain interpretable (standard IHC practice). |
| Only faint staining appears in thyroid or colon glandular cells. | HPA assigns those populations a low staining level (HPA tissue IHC). Faint signal there alone does not show that the run failed. | Judge run performance using a reported medium staining population and appropriate controls before changing assay conditions (HPA tissue IHC; standard IHC practice). |
| Signal appears mainly along cell edges. | ARHGEF7 can localise to the cortex and cell junctions, but HPA describes the overall tissue profile as cytoplasmic (UniProt Q14155; HPA tissue IHC). | Check for accompanying cytoplasmic staining and compare control sections; avoid calling a uniform membrane outline a confirmed target pattern (standard IHC practice; UniProt Q14155). |
Experiment evidence
IHC & IF images showing ARHGEF7 staining
A02764-2 · IHCIHC analysis of ARHGEF7 using anti-ARHGEF7 antibody (A02764-2). ARHGEF7 was detected in a paraffin-embedded section of human liver cancer ti
A02764-2 · IHCIHC analysis of ARHGEF7 using anti-ARHGEF7 antibody (A02764-2). ARHGEF7 was detected in a paraffin-embedded section of human liver cancer ti
A02764-2 · IFIF analysis of ARHGEF7 using anti-ARHGEF7 antibody (A02764-2). ARHGEF7 was detected in an immunocytochemical section of HELA cells. Enzyme a
A02764 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200(4° overnight). 2, Tris-EDTA,pH9.0 was used
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000102606-ARHGEF7/tissue/cerebral+cortex#imid_1501416
HPA · Small intestineHuman Protein Atlas IHC, Small intestine. Source: https://www.proteinatlas.org/ENSG00000102606-ARHGEF7/tissue/small+intestine#imid_1501333
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000102606-ARHGEF7/tissue/colon#imid_1501342
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000102606-ARHGEF7/tissue/liver#imid_1501359
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ARHGEF7 IHC & IF
🧪Run adipose tissue first and require staining in adipocytes (HPA: Medium in adipocytes). Use salivary gland glandular cells as the negative tissue (HPA: Not detected); on the positive slide, score other cells as internal negatives only if their identity and lack of ARHGEF7 expression are independently established, and expect background-level signal in those cells.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Salivary gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ARHGEF7; derive a cell-line control from the positive tissue's cell type (Adipocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a rabbit IgG isotype control matched to the primary antibody’s clonality and concentration (selected-SKU caption: rabbit primary); and an ARHGEF7 knockout control processed alongside the test sections. Quench endogenous peroxidase and inspect background in adipose tissue before interpreting DAB staining (selected-SKU caption: peroxidase detection with DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A02764-2 tissue-IHC caption does not report a fixative (selected-SKU caption: fixative not stated). The demonstrated paraffin-section procedure uses heat retrieval in EDTA at pH 8.0; that caption does not establish whether retrieval is essential (selected-SKU caption: EDTA heat retrieval). Frozen sections or IF cannot be judged easier from the supplied evidence, and HPA lists no ICC-IF images (HPA subcellular: no ICC-IF cell lines); in paraffin adipose sections, lipid extraction can leave adipocyte vacuoles, so assess staining at the cell rim.
HPA tissue IHC evidence for ARHGEF7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced ARHGEF7 IHC Tips
Troubleshoot ARHGEF7 chromogenic IHC in paraffin sections by checking retrieval, staining controls, compartment, and cell type before interpreting signal (datasheet A02764-2; HPA tissue IHC).
What retrieval should I try first when ARHGEF7 staining is weak?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A02764-2). The selected tissue image used this retrieval before 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A02764-2). If staining remains weak, compare heating times on adjacent sections while holding antibody concentration and DAB development constant, and inspect morphology for damage (standard IHC practice). Score cytoplasmic staining separately from tissue edge color, and include a no primary control to detect nonspecific signal that increases with retrieval (HPA: ubiquitous cytoplasmic expression; standard IHC practice).
How can I distinguish fixation effects from weak ARHGEF7 staining?
ARHGEF7 fixation sensitivity is unknown: the selected paraffin tissue caption does not report a fixative, and no target specific fixation comparison is supplied (datasheet A02764-2). Record fixation and processing history, then compare adjacent sections from one block using the same EDTA pH 8.0 retrieval and detection settings (standard IHC practice; datasheet A02764-2). If multiple samples from one processing batch lose cytoplasmic staining, compare retrieval times while watching tissue integrity and background (HPA: ubiquitous cytoplasmic expression; standard IHC practice). Phosphorylation sites and the absence of a transmembrane segment do not establish a fixation response (UniProt Q14155).
Should ARHGEF7 appear at cell borders or throughout the cytoplasm?
Expect a cytoplasmic pattern, potentially enriched toward the cortex or cell junctions; UniProt also places ARHGEF7 at ruffles, lamellipodia, and adhesions (HPA: ubiquitous cytoplasmic expression; UniProt Q14155 subcellular location). Only a small proportion is reported at focal adhesions, so weak border accentuation need not dominate a chromogenic section (UniProt Q14155 subcellular location). Compare central and edge regions of intact cells at the same magnification, with matched exposure and DAB development across sections (standard IHC practice). Diffuse nuclear only color or staining restricted to damaged edges warrants review of controls before assigning ARHGEF7 localisation (UniProt Q14155 subcellular location; standard IHC practice).
Could isoforms or epitope accessibility explain inconsistent staining?
ARHGEF7 has 6 annotated isoforms, but the supplied antibody caption does not identify its epitope or establish isoform coverage (UniProt Q14155 isoforms; datasheet A02764-2). Its CH, SH3, DH, and PH domains span different sequence regions, so an antibody directed to one region could have different isoform coverage; verify the actual immunogen before making that inference for this antibody (UniProt Q14155 domains and isoforms; standard antibody validation practice). Compare staining across matched sections using the same EDTA pH 8.0 retrieval (datasheet A02764-2; standard IHC practice). Phosphorylation is documented, but these data do not show whether it changes antibody binding (UniProt Q14155 modified residues; datasheet A02764-2).
How should I assess ARHGEF7 by IF alongside this IHC result?
Treat IF/ICC as a separate validation experiment; the selected antibody evidence describes paraffin section chromogenic IHC, without an IF/ICC image or fixation result (datasheet A02764-2; HPA subcellular). Multiplex with a validated marker for the expected cell population, such as a glandular population where tissue IHC reports ARHGEF7, and check cell level overlap (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores, favoring a longer wavelength channel if the specimen shows autofluorescence, and include single color controls (standard IF practice). Because ARHGEF7 lacks a transmembrane segment and has cytoplasmic and cortical locations, optimize permeabilisation for access to an intracellular epitope after confirming the antibody epitope (UniProt Q14155 topology and subcellular location; standard IF practice).
What causes widespread brown staining in ARHGEF7 sections?
The selected image used 10% goat serum blocking, 2 μg/ml rabbit primary overnight at 4°C, and an HRP secondary followed by DAB (datasheet A02764-2). Check a no primary section for secondary reagent binding and endogenous enzyme color, then apply a peroxidase block as part of the general chromogenic workflow (standard IHC practice). If the control is clean, titrate primary concentration or shorten DAB development while retaining EDTA pH 8.0 retrieval (datasheet A02764-2; standard IHC practice). Interpret residual cytoplasmic color against cell morphology because broad cytoplasmic expression is reported across tissues (HPA tissue IHC; standard IHC practice).
How should I quantify ARHGEF7 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Predefine the cell population and viable tissue area, then score cytoplasmic intensity as 0–3 and calculate an H-score from intensity weighted percentages, with a possible range of 0–300 (HPA: ubiquitous cytoplasmic expression; standard IHC scoring practice). Alternatively report the percentage of positive cells or positive cell density per mm² when cell segmentation is reliable (standard IHC scoring practice). Normalize comparisons to the same eligible cell population or viable area, and hold retrieval, antibody incubation, imaging, and DAB development constant (standard IHC practice). Report stromal and glandular compartments separately where applicable, since HPA describes staining by cell type and rates antibody versus RNA agreement as medium (HPA tissue IHC).
How can I distinguish true ARHGEF7 staining from artefact?
A credible result is cytoplasmic staining in intact cells, with possible cortical accentuation; focal adhesion staining may represent only a small component (HPA: ubiquitous cytoplasmic expression; UniProt Q14155 subcellular location). Compare the same cell population across sections and inspect nuclei, tissue edges, and necrotic areas before calling a positive result (standard IHC practice). Staining confined to edges or damaged tissue, or color persisting without primary antibody, supports artefact or endogenous enzyme activity rather than target assignment (standard IHC practice). HPA reports glandular cells in salivary gland as not detected, but its tissue IHC reliability is Approved with medium antibody versus RNA consistency, so use that comparison cautiously (HPA tissue IHC).
Boster reagents
Best ARHGEF7 / Rho guanine nucleotide exchange factor 7 IHC Antibodies
Both antibodies have human paraffin-section IHC images; A02764-2 also has IF images in HeLa cells and human colon cancer tissue (catalog image captions). Human, mouse and rat reactivity is listed for both (catalog).
Real IHC data
Anti-ARHGEF7 Antibody ®
Cat # A02764-2
Real IHC data
Anti-ARHGEF7/Beta Pix Antibody
Cat # A02764
A02764 has an IHC image of human paraffin-embedded tonsil using Tris-EDTA retrieval at pH 9.0 (A02764 image caption). A02764-2 has IHC images of human paraffin-embedded liver cancer and lung adenocarcinoma using EDTA retrieval at pH 8.0, plus IF images of HeLa cells and human colon cancer tissue (A02764-2 image captions).
Which to pick: For tissue IHC, choose A02764 for the documented tonsil protocol at 1:200 (A02764 image caption), or A02764-2 for its documented liver cancer and lung adenocarcinoma protocols at 2 μg/ml (A02764-2 image captions); both captions specify paraffin sections but leave the fixative unreported. For IF/ICC, A02764-2 has a documented HeLa cell IF image at 5 μg/ml (A02764-2 IF image caption), while A02764 lists IF and ICC applications without an IF image in the payload (A02764 catalog). For mouse or rat work, both list reactivity with those species (catalog), but the supplied IHC images document human tissue only (catalog image captions); A02764 is identified as polyclonal (A02764 catalog), while A02764-2 has no clonality stated in the payload.
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14155 (ARHG7_HUMAN, Rho guanine nucleotide exchange factor 7).
- Human Protein Atlas. ARHGEF7 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ARHGEF7 subcellular location (ICC-IF): Highest expression in JURKAT: 75.7 nTPM.
- Human Protein Atlas. ARHGEF7 antibody validation summary (1 antibodies).
- ARHGEF7 promotes metastasis of colorectal adenocarcinoma by regulating the motility of cancer cells. International journal of oncology 2018 — PMC6192735.
- Arhgef7 is essential for granule cell precursor proliferation and migration during cerebellum development. iScience 2026 — PMC12828525.
- PubMed PMID:8590280 — UniProt-cited evidence.
- PubMed PMID:9726964 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ARHGEF7 (Rho guanine nucleotide exchange factor 7). Expected localisation Cytoplasmic tissue signal (HPA tissue IHC); adhesion-associated protein (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02764-2). Validated IHC antibody: A02764-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Salivary gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A02764-2); 2. Fixation: Image fixative and duration unreported (datasheet A02764-2); 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 A02764-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02764-2); 8. Primary antibody: Rabbit anti-ARHGEF7, 2-5 μg/ml (datasheet A02764-2); 9. Primary incubation: Overnight at 4 °C (datasheet A02764-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A02764-2); verify before use.; Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 6 isoforms; epitope coverage needs checking (UniProt); Regulation: Expression regulation not established (UniProt). Evidence: UniProt Q14155, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in JURKAT: 75.7 nTPM, Datasheet A02764-2, PMC6192735. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ARHGEF7/immunohistochemistry-ihc-guide.