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IQGAP1 / Ras GTPase-activating-like protein IQGAP1 · IHC design guide
Design Immunohistochemistry for IQGAP1
Plan chromogenic IQGAP1 IHC in paraffin sections with the catalog antibody (datasheet A01603). Compare cytoplasmic and membranous staining across cell types, using strongly stained colon glandular cells and undetected adipocytes as reference populations (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IQGAP1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Most cell types show cytoplasmic and membranous signal (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01603) | |
| Positive control | Colon+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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Macrophages can contribute strong lung staining (HPA tissue IHC) | |
| Regulation | Nuclear levels rise at G1/S (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; chain spans residues 2–1657 (UniProt) |
Section 1
Recommended IQGAP1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by four published IQGAP1 chromogenic IHC protocols covering melanoma, colorectal carcinoma, hepatocellular carcinoma, and lung cancer (PMC5406000; PMC5588162; PMC9628040; PMC4249885).
| Sample | Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet A01603) |
| Fixation | Image fixative and duration unreported (datasheet A01603); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01603) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01603) |
| Primary antibody | Rabbit anti-IQGAP1, 0.5-1μg/ml (datasheet A01603) |
| Primary incubation | Overnight at 4 °C (datasheet A01603) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01603) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | IQGAP1-positive staining in glandular cells of colon (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in most cell types. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 for paraffin sections (datasheet A01603); the colorectal, hepatocellular, and lung protocols also report citrate retrieval (PMC5588162; PMC9628040; PMC4249885).
Section 2
What Is the Expected IQGAP1 Staining Pattern?
In paraffin sections, expect cytoplasmic and membrane-associated IQGAP1 staining in many cell types, with high staining in selected populations such as colon glandular cells and lung macrophages (HPA tissue IHC: Enhanced; HPA: High in colon glandular cells and lung macrophages). IQGAP1 also localizes to the cell cortex and can enter the nucleus; it has no transmembrane segment (UniProt P46940: subcellular location and topology).
| Brown cytoplasmic and membrane-associated staining in colon glandular cells or lung macrophages. | This fits the reported IHC pattern and provides a useful positive reference; compare the identified cells, rather than judging the entire section by one average intensity (HPA tissue IHC: cytoplasmic and membranous expression; HPA: High in these cells). |
| Signal is mainly in luminal material or extracellular spaces, with little staining in cells. | That distribution does not match the reported cellular pattern; inspect morphology and reagent controls before scoring it as IQGAP1 (HPA tissue IHC: cytoplasmic and membranous expression). Nuclear staining alone needs context because nuclear IQGAP1 is reported (UniProt P46940: nucleus). |
| Strong staining appears in adipocytes or skeletal myocytes. | These cells are reported as not detected; check for cross-reactivity or endogenous detection activity before accepting the signal (HPA tissue IHC: adipocytes and skeletal myocytes, Not detected). Their status does not make every cell in those tissues a negative control. |
| A similar brown haze covers cells, stroma and areas without tissue. | This is more consistent with background than a cell-localized result. Review the reagent-only control, blocking, washing and chromogen development as general chromogenic IHC checks; the expected IQGAP1 pattern is cellular (HPA tissue IHC: cytoplasmic and membranous expression). |
| Colon glandular cells or lung macrophages show little or no signal. | A negative result in these reported high-staining cells warrants a run-level check before biological interpretation (HPA tissue IHC: High in colon glandular cells and lung macrophages). Confirm cell identity, positive-control performance and that the IHC-validated antibody was used. |
💡Expected IQGAP1 appearanceCall a result positive when identified colon glandular cells or lung macrophages show clear cytoplasmic and membrane-associated chromogen with high staining; diffuse extracellular brown deposit is suspect (HPA tissue IHC: pattern and High in these cells).
| Cellular location | IQGAP1 is reported at the cortex, apical and basolateral membranes, cytoplasm and nucleus, despite having no transmembrane segment (UniProt P46940: subcellular location and topology). A membrane-associated outline can therefore be plausible without implying that the protein spans the membrane. |
| Cell cycle | Nuclear levels increase at G1/S (UniProt P46940: subcellular distribution). Evaluate nuclear signal alongside cell identity and the broader cytoplasmic or membrane pattern; nuclear staining by itself is not sufficient grounds to call the section an artefact. |
| Choice of comparison cells | Colon glandular cells, kidney glomerular cells, lung macrophages and placental trophoblastic cells are reported as High; adipocytes and skeletal myocytes are Not detected (HPA tissue IHC). Score the named cells, since each designation applies to a cell population. |
| Evidence for the tissue pattern | The tissue IHC profile is rated Enhanced for consistency between staining and RNA data; HPA014055 and CAB013302 each have Enhanced IHC validation (HPA tissue IHC: reliability; HPA antibodies: IHC). Use this as support for the reported pattern, not as proof of specificity in every section. |
| Chromogenic detection | Endogenous enzyme activity can imitate antibody-dependent color in chromogenic IHC (general IHC practice). Where relevant to the detection system, evaluate an appropriate reagent control and enzyme block; HPA cell-level observations alone cannot identify the source of an unexpected deposit. |
| Q: Can ICC-IF resolve an IHC staining call? | A: ICC-IF supports plasma-membrane and cell-junction localization, and CAB013302 has Supported ICC validation (HPA subcellular; HPA antibodies: ICC). This is useful localization context; decide paraffin-section staining against the tissue IHC pattern (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in a positive reference section. | A failed staining run or unsuitable assay settings are possible; the selected cells are reported as High (HPA tissue IHC: colon glandular cells; lung macrophages). | Confirm the named cells are present and the IHC-validated antibody and detection reagents were used; review retrieval, reagent preparation and positive-control performance as general IHC practice. |
| Brown deposit sits mainly outside cells. | The distribution conflicts with the reported cytoplasmic and membranous cellular pattern (HPA tissue IHC: profile). | Inspect the counterstain and tissue boundaries, then compare reagent controls and chromogen development. Score only convincing cell-associated signal (general IHC practice; HPA tissue IHC: profile). |
| Adipocytes or skeletal myocytes stain strongly. | Cross-reactivity or endogenous detection activity is possible because these named cells are reported as Not detected (HPA tissue IHC). | Check a reagent-only control and the relevant endogenous-enzyme block; compare another reported cell population before assigning target-specific staining (general IHC practice; HPA tissue IHC: cell-level pattern). |
| Most of the slide has a uniform brown haze. | Nonspecific reagent binding, inadequate washing or excess chromogen development can produce background (general IHC practice). | Review blocking, washing and development time against the run controls; require a recognizable cytoplasmic or membrane-associated cell pattern for interpretation (general IHC practice; HPA tissue IHC: profile). |
| Some cells in a positive tissue stain while neighbors do not. | The HPA levels describe specific populations, such as kidney glomerular cells and lung macrophages, rather than every cell in a tissue (HPA tissue IHC). | Identify and score the reported population separately. Compare intensity only after matching cell type and section morphology (HPA tissue IHC: cell-level observations; general IHC practice). |
| Nuclear color is present with weak membrane staining. | Nuclear IQGAP1 is plausible, and its level can rise at G1/S; that fact alone does not establish that this particular nuclear signal is specific (UniProt P46940: subcellular distribution). | Check whether color follows intact nuclei, review reagent controls and look for a credible cellular pattern in reported positive cells before scoring (general IHC practice; HPA tissue IHC: positive cells). |
Experiment evidence
IHC & IF images showing IQGAP1 staining
A01603 · IHCIHC analysis of IQGAP1 using anti-IQGAP1 antibody (A01603). IQGAP1 was detected in paraffin-embedded section of human intestinal cancer tiss
A01603 · IHCIHC analysis of IQGAP1 using anti-IQGAP1 antibody (A01603). IQGAP1 was detected in paraffin-embedded section of human intestinal cancer tiss
A01603 · IFIF analysis of IQGAP1 using anti-IQGAP1 antibody (A01603). IQGAP1 was detected in immunocytochemical section of U20S cells. Enzyme antigen r
A01603-1 · IHCIHC analysis of IQGAP1 using anti-IQGAP1 antibody (A01603-1). IQGAP1 was detected in a paraffin-embedded section of human endometrial adenoc
A01603-1 · IHCIHC analysis of IQGAP1 using anti-IQGAP1 antibody (A01603-1). IQGAP1 was detected in a paraffin-embedded section of human ovarian cancer tis
A01603-1 · IFIF analysis of IQGAP1 using anti-IQGAP1 antibody (A01603-1). IQGAP1 was detected in immunocytochemical section of A431 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000140575-IQGAP1/tissue/cerebral+cortex#imid_3677080
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000140575-IQGAP1/tissue/colon#imid_3676990
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000140575-IQGAP1/tissue/liver#imid_3677007
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000140575-IQGAP1/tissue/kidney#imid_3677000
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB013302; green = IQGAP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB013302; green = IQGAP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IQGAP1 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue as the negative comparator, scoring adipocytes (HPA: Not detected in adipocytes); within the colon slide, cells without specific staining can serve as internal background comparators, but no other colon cell type is established as negative by the supplied rows.
Positive control tissue: Colon (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show IQGAP1 in A-431, U-251MG, U2OS, with annotated localisation: Plasma membrane (supported), Cell Junctions (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide; a host- and clonality-matched rabbit IgG control (caption: rabbit primary antibody); and IQGAP1-knockout tissue if available, or peptide-blocked primary antibody when a matching peptide is available. Quench endogenous peroxidase and check for endogenous biotin background in colon, since the selected caption uses biotin-based detection and DAB (A01603 tissue-IHC caption).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01603 paraffin-section caption does not report a fixative. That caption documents heat retrieval in citrate buffer at pH 6 for 20 minutes, but does not establish whether retrieval is required (A01603 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; with the caption’s biotin-based DAB method, assess endogenous biotin background before scoring colon glands (A01603 tissue-IHC caption).
HPA tissue IHC evidence for IQGAP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IQGAP1 IHC Tips
Troubleshoot IQGAP1 staining by checking retrieval, tissue preservation, compartment patterns, controls, and cell-specific scoring before interpreting signal.
How should I retrieve IQGAP1 in paraffin sections when staining is weak?
Use heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes as the starting condition for A01603 (datasheet A01603; A01603 tissue-IHC caption). The caption reports 1 μg/ml primary antibody incubated overnight at 4°C, so compare retrieval changes while holding that incubation constant (A01603 tissue-IHC caption). If signal remains weak, compare a longer heat exposure on matched sections while watching for tissue damage (standard IHC practice). Include a known positive compartment, such as colon glandular cells, and a primary-antibody omission control in each run (HPA: High in colon glandular cells; standard IHC practice).
Could fixation explain weak or uneven IQGAP1 staining?
The A01603 tissue-IHC caption identifies a paraffin section but does not state its fixative; target-specific sensitivity to fixation is therefore unknown (A01603 tissue-IHC caption). Record fixative, fixation duration, section age, and processing history for each specimen before comparing staining intensity (standard IHC practice). Test matched sections with citrate retrieval at pH 6 for 20 minutes, holding antibody concentration at 1 μg/ml where appropriate for A01603 (A01603 tissue-IHC caption). Compare internal cell populations and tissue morphology across sections, and treat a fixation effect as a working explanation only if it tracks the documented processing differences (standard IHC practice).
Which subcellular pattern should count as plausible IQGAP1 staining?
Expect predominantly cytoplasmic and membranous staining across many cell types in tissue sections (HPA: tissue-IHC profile). Plasma membrane and cell-junction localisation are supported by cell imaging; UniProt also lists cell cortex, apical and basolateral membrane, cytoplasm, and nucleus (HPA: subcellular summary; UniProt P46940 subcellular location). A sharply nuclear-only pattern deserves independent confirmation, although nuclear IQGAP1 can rise at the G1/S transition (UniProt P46940 subcellular location, PubMed:20883816). Compare compartments within the same cell population, confirm preserved morphology with the counterstain, and review omission controls before assigning diffuse edge staining to IQGAP1 (standard IHC practice).
How do epitope location and processing affect an unexpected staining pattern?
The supplied record lists 0 isoforms, no transmembrane segment, and a protein chain spanning residues 2–1657 (UniProt P46940 isoforms, topology, processing). IQGAP1 has CH, WW, IQ, and Ras-GAP regions plus reported modified residues, so an antibody’s mapped immunogen is useful when assessing epitope accessibility (UniProt P46940 domains and modified residues; standard IHC practice). The A01603 caption does not map its epitope, so staining alone cannot identify which region the antibody recognizes (A01603 tissue-IHC caption). Check the antibody’s epitope documentation, compare positive and low-expression cell populations on matched sections, and seek an independently mapped antibody when specificity remains uncertain (standard IHC practice; HPA: tissue-IHC profile).
How should I investigate IQGAP1 localisation with multiplex IF?
Use IF as a complementary check of the membrane and cell-junction pattern supported by cell imaging, while scoring chromogenic tissue IHC separately (HPA: subcellular summary; standard IHC practice). Pair IQGAP1 with a marker of the cell population under study; lung macrophages or colon glandular cells are documented high-expression examples (HPA: High in lung macrophages and colon glandular cells). Choose spectrally separated fluorophores and a longer-wavelength IQGAP1 channel when tissue autofluorescence interferes, then inspect single-stain controls (standard IF practice). Because IQGAP1 has no transmembrane segment and can occupy cytoplasmic and cortical compartments, optimise permeabilisation against the antibody’s mapped epitope and confirm that membrane outlines remain intact (UniProt P46940 topology and subcellular location; standard IF practice).
What should I change when DAB staining obscures cellular detail?
First compare the primary-antibody omission control with the stained section to separate detection-system background from antibody-dependent signal (standard IHC practice). The A01603 example used 10% goat serum, biotinylated secondary antibody, a streptavidin–biotin complex, and DAB, making blocking and detection controls directly relevant to that workflow (A01603 tissue-IHC caption). Apply a peroxidase block and check whether DAB precipitate, endogenous biotin, or insufficient washing explains staining outside intact cells (standard chromogenic IHC practice). If background remains antibody-dependent, titrate around the reported 1 μg/ml primary concentration and reassess overnight incubation at 4°C on matched sections (A01603 tissue-IHC caption; standard IHC practice).
How should I score IQGAP1 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because tissue IHC commonly shows cytoplasmic and membranous IQGAP1 (HPA: tissue-IHC profile). For identifiable cells, record the percentage positive and an H-score from intensity categories 0–3; report membrane and cytoplasmic scores separately when feasible (standard IHC scoring practice). For dispersed positive cells, report positive-cell density per mm² of viable tissue and specify the counted region (standard IHC scoring practice). Normalise to the number of evaluable cells or viable tissue area, use the same threshold and imaging settings across groups, and document excluded necrotic or damaged regions (standard IHC scoring practice).
How can I distinguish convincing IQGAP1 signal from tissue artefact?
A convincing result follows cellular boundaries or cytoplasm in an intact, identifiable population and agrees with appropriate controls (HPA: tissue-IHC profile; standard IHC practice). Colon glandular cells and lung macrophages are high-expression references, whereas adipocytes are listed as not detected; evaluate each population rather than treating every cell in a section as equivalent (HPA: colon, lung, and adipose tissue-IHC entries). Predominantly extracellular deposit, a bright cut edge, necrotic material, or signal in a primary-antibody omission control raises concern for artefact (standard IHC practice). Investigate isolated nuclear-only or diffuse staining against morphology and controls, while allowing for reported cell-cycle-dependent nuclear localisation (UniProt P46940 subcellular location, PubMed:20883816; standard IHC practice).
Boster reagents
Best IQGAP1 / Ras GTPase-activating-like protein IQGAP1 IHC Antibodies
Two anti-IQGAP1 antibodies have paraffin-section IHC images from human cancers and IF/ICC images from cells (catalog image captions); catalog reactivity covers human, mouse and rat (catalog: A01603, A01603-1).
Real IHC data
Anti-IQGAP1 Antibody ®
Cat # A01603
Real IHC data
Anti-IQGAP1 Antibody ®
Cat # A01603-1
A01603 will render with human intestinal cancer IHC and U20S cell IF/ICC images; its listed reactivity is human, mouse and rat (catalog: A01603 image captions and reactivity). A01603-1 will render with human endometrial adenocarcinoma IHC and A431 cell IF/ICC images; its listed reactivity is human (catalog: A01603-1 image captions and reactivity).
Which to pick: For paraffin-section IHC, start with A01603 at 0.5–1 μg/ml with citrate retrieval at pH 6, or A01603-1 at 2–5 μg/ml with EDTA retrieval at pH 8; each retrieval method comes from that SKU’s own IHC image caption (catalog: A01603 and A01603-1 IHC captions and dilutions). For IF/ICC, both list the application and have cell images: A01603 at 2 μg/ml or A01603-1 at 5 μg/ml (catalog: IF/ICC applications, IF image captions and dilutions). Choose A01603 when mouse or rat reactivity is needed, while treating that as listed reactivity rather than tissue IHC image evidence; clone type is unreported for both antibodies, and neither IHC caption reports the tissue fixative (catalog: reactivity, clone fields and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P46940 (IQGA1_HUMAN, Ras GTPase-activating-like protein IQGAP1).
- Human Protein Atlas. IQGAP1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. IQGAP1 subcellular location (ICC-IF): Localized to the plasma membrane and cell junctions..
- Human Protein Atlas. IQGAP1 antibody validation summary (2 antibodies).
- IQGAP1 is an oncogenic target in canine melanoma. PloS one 2017 — PMC5406000.
- Alterations in IQGAP1 expression and localization in colorectal carcinoma and liver metastases following oxaliplatin-based chemotherapy. Oncology letters 2017 — PMC5588162.
- Comprehensive analysis of the expression and prognosis for IQ motif-containing GTPase-activating proteins in hepatocellular carcinoma. BMC cancer 2022 — PMC9628040.
- Coexpression of IQ-domain GTPase-activating protein 1 (IQGAP1) and Dishevelled (Dvl) is correlated with poor prognosis in non-small cell lung cancer. PloS one 2014 — PMC4249885.
- PubMed PMID:8051149 — UniProt-cited evidence.
- PubMed PMID:7584044 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IQGAP1 (Ras GTPase-activating-like protein IQGAP1). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01603). Validated IHC antibody: A01603 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Colon (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet A01603); 2. Fixation: Image fixative and duration unreported (datasheet A01603); 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: Citrate pH 6 , 20 min (datasheet A01603); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01603); 8. Primary antibody: Rabbit anti-IQGAP1, 0.5-1μg/ml (datasheet A01603); 9. Primary incubation: Overnight at 4 °C (datasheet A01603). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Macrophages can contribute strong lung staining (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; chain spans residues 2–1657 (UniProt); Regulation: Nuclear levels rise at G1/S (UniProt). Evidence: UniProt P46940, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane and cell junctions., Datasheet A01603, PMC5406000, PMC5588162. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IQGAP1/immunohistochemistry-ihc-guide.