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DLC1 / Rho GTPase-activating protein 7 · IHC design guide
Design Immunohistochemistry for DLC1
Plan chromogenic IHC on paraffin sections with the catalog antibody at 1:100–1:300 (datasheet). Assess membranous and luminal staining in glandular cells, keeping in mind the uncertain tissue staining reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DLC1 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 | Membranous and luminal tissue staining (HPA tissue IHC) | |
| Staining pattern | Membranous and luminal staining in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Gallbladder+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 paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Staining has low agreement with RNA; verification is pending (HPA tissue IHC) | |
| Regulation | Expression regulation is not established (UniProt) | |
| Isoform / epitope | 6 isoforms; antibody epitope coverage is unknown (UniProt) |
Section 1
Recommended DLC1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with 3 published DLC1 staining protocols for paraffin sections.
| Sample | Paraffin-embedded human prostate carcinoma tissue; fixative not specified (datasheet A30552) |
| Fixation | Image fixative and duration unreported (datasheet A30552); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-DLC1, 1:100 - 1:300 (datasheet A30552) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DLC1-positive staining in glandular cells of gallbladder (HPA tissue IHC: High). HPA tissue profile: Membranous and luminal expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); the published protocols show alternative retrieval conditions.
Section 2
What Is the Expected DLC1 Staining Pattern?
DLC1 is a cytoplasmic protein associated with focal adhesions and membranes, with no transmembrane segment (UniProt Q96QB1). In paraffin-section IHC, look for signal in gallbladder glandular cells and, at lower reported intensity, colon and small-intestine glandular cells or kidney tubule cells (HPA tissue IHC). HPA describes membranous and luminal staining across several tissues, but rates its tissue IHC profile Uncertain because staining and RNA data have low consistency (HPA tissue IHC).
| Glandular cells show localized staining in gallbladder, with weaker staining in colon or small intestine. | This follows HPA's High gallbladder and Medium colon and small-intestine observations (HPA tissue IHC). Assess cell identity and localization together; those observations remain provisional because the tissue IHC profile is Uncertain (HPA tissue IHC). |
| Signal appears at cell edges or in cytoplasm, sometimes alongside luminal staining. | Cytoplasmic and focal-adhesion-associated localization is consistent with UniProt Q96QB1; HPA reports membranous and luminal tissue staining (HPA tissue IHC). Luminal signal alone deserves scrutiny: HPA's pattern has not received strong tissue-level validation (HPA tissue IHC). |
| Nuclear staining dominates, with little signal in the reported-positive cells. | A dominant nuclear pattern conflicts with the supplied cytoplasmic, focal-adhesion and membrane annotations (UniProt Q96QB1). Treat it as a suspected nonspecific or technical signal and check controls before assigning it to DLC1 (general IHC practice). |
| Reported-negative cells stain as strongly as the expected glandular cells. | HPA reports DLC1 as Not detected in adipocytes of adipose tissue and in adrenal glandular cells (HPA tissue IHC). Strong staining there raises concern about cross-reactivity or endogenous detection activity; it does not alone disprove expression, given the Uncertain HPA profile (HPA tissue IHC; general IHC practice). |
| The whole section is diffusely colored, or a reported-positive tissue has no discernible cellular signal. | Uniform color obscures compartment and cell-type assessment, while absent signal cannot distinguish a technical failure from an antibody or specimen difference (general IHC practice). Compare controls and the complete cellular pattern before interpreting either result (general IHC practice). |
💡Expected DLC1 appearanceA plausible positive is localized cytoplasmic or cell-edge staining in gallbladder glandular cells at the reported High level; widespread nuclear or uniform noncellular color is suspect (UniProt Q96QB1; HPA tissue IHC; general IHC practice).
| How firm is the tissue reference pattern? | HPA calls tissue IHC Uncertain because antibody staining and RNA expression have low consistency; the supplied IHC antibody entry HPA017753 is also Uncertain (HPA tissue IHC; HPA antibodies). Use reported intensities as comparison points, not absolute pass or fail criteria (general IHC practice). |
| Which compartment should guide scoring? | UniProt places DLC1 in cytoplasm, focal adhesions and membrane-associated regions, without a transmembrane segment (UniProt Q96QB1). HPA tissue IHC reports membranous and luminal expression (HPA tissue IHC). Record the exact compartment seen rather than treating all edge or luminal color as equivalent (general IHC practice). |
| Can one stain represent every isoform? | UniProt lists 6 DLC1 isoforms (UniProt Q96QB1). Antibody epitope coverage is not supplied, so an isoform-specific staining prediction is unavailable; interpret an absent signal cautiously when comparing reagents (general IHC practice). |
| What does IF/ICC add? | HPA ICC-IF places DLC1 mainly in supported cytosol, with approved additional endoplasmic-reticulum and vesicle locations (HPA subcellular). This answers the localization question for IF/ICC; its cell images and supported ICC antibody entries do not validate the uncertain tissue IHC pattern (HPA subcellular; HPA antibodies; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Gallbladder glandular cells show no signal. | HPA reports High staining there, but its tissue IHC assessment is Uncertain; a technical failure or reagent-dependent result is possible (HPA tissue IHC; general IHC practice). | Check tissue preservation, the detection control and the antibody's supplied IHC-P instructions; repeat with a working control before calling the sample negative (general IHC practice). |
| Only nuclei stain strongly. | Nuclear dominance does not fit the supplied DLC1 localization (UniProt Q96QB1). Nonspecific antibody or detection signal is possible (general IHC practice). | Review the no-primary control and compare nuclear color with cytoplasmic and cell-edge signal in reported-positive cells (general IHC practice). |
| Adipocytes or adrenal glandular cells stain strongly. | Those cells are reported Not detected by HPA tissue IHC, although the overall profile is Uncertain (HPA tissue IHC). Cross-reactivity or endogenous detection activity may contribute (general IHC practice). | Run a no-primary control and check detection blocking; compare the same run with reported-positive glandular tissue before interpreting specificity (general IHC practice). |
| Diffuse color obscures cell boundaries. | Background from antibody or detection reagents can make a cellular pattern unreadable (general IHC practice). | Check the no-primary control, blocking, washes and reagent concentration under the chosen IHC-P workflow; score only interpretable cellular signal (general IHC practice). |
| Luminal color is prominent but glandular cells are faint. | HPA reports luminal staining, yet the tissue IHC profile has low consistency with RNA data (HPA tissue IHC). Luminal color alone is therefore weak evidence for DLC1-positive cells (HPA tissue IHC; general IHC practice). | Document luminal and cellular staining separately; compare reported-positive and reported-negative tissues plus the no-primary control (HPA tissue IHC; general IHC practice). |
| Two antibodies give different tissue patterns. | The supplied HPA tissue IHC entry is Uncertain, and the listed antibodies have different IHC and ICC validation entries (HPA tissue IHC; HPA antibodies). Epitope coverage is unspecified (HPA antibodies). | Keep each antibody's observations separate, compare localization and controls, and avoid declaring either pattern validated from agreement with HPA intensity alone (general IHC practice; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing DLC1 staining
A30552 · IHCImmunohistochemistry analysis of paraffin-embedded human prostate carcinoma tissue, using RHG07 Antibody. The picture on the right is blocke
A30552 · IFImmunofluorescence analysis of A549 cells, using RHG07 Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000164741-DLC1/tissue/cerebral+cortex#imid_4705728
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000164741-DLC1/tissue/colon#imid_4705662
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000164741-DLC1/tissue/liver#imid_4705671
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000164741-DLC1/tissue/kidney#imid_4705664
![HPA ICC-IF BJ [Human fibroblast] — DLC1 (HPA061655)](https://images.proteinatlas.org/61655/1801_B4_4_cr59ca0d3dc6143_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA061655; green = DLC1, blue = nucleus, red = microtubules.
![HPA ICC-IF BJ [Human fibroblast] — DLC1 (HPA061655)](https://images.proteinatlas.org/61655/1801_B4_6_cr59c4f4e721d1a_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA061655; green = DLC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DLC1 IHC & IF
🧪Run gallbladder first: its glandular cells must show DLC1 staining (HPA: High in gallbladder glandular cells). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); on the gallbladder slide, assess non-glandular cells for background signal, but do not assume they are validated target-negative cells (HPA: level reported for glandular cells only).
Positive control tissue: Gallbladder (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DLC1 in BJ [Human fibroblast], SuSa, U2OS, THP-1, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a host-species- and immunoglobulin-class-matched isotype control where applicable, and a DLC1 knockout biological negative if available; the catalog antibody’s peptide-blocked prostate carcinoma image provides an additional competition control (selected tissue-IHC caption: peptide block). For chromogenic gallbladder IHC, block endogenous peroxidase and, if using avidin–biotin detection, check endogenous biotin (standard IHC practice).
⚠️Feasibility: A DLC1-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; optimize retrieval empirically (supplied target/application evidence: no fixation window or retrieval conditions). The selected A30552 paraffin-section caption does not report a fixative (selected tissue-IHC caption: fixative not stated). ICC/IF has image evidence in BJ, SuSa, U2OS and THP-1, but the evidence does not establish that IF or frozen sections are easier than paraffin IHC (HPA subcellular: ICC-IF image cell lines); assess gallbladder background with the detection controls (standard IHC practice).
HPA tissue IHC evidence for DLC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DLC1 IHC Tips
Troubleshoot DLC1 staining in paraffin sections by checking retrieval, compartment, cell type and controls before interpreting chromogenic signal.
What retrieval conditions should I start with for DLC1 in paraffin sections?
Use citrate buffer at pH 6.0 for heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). Let slides cool in buffer for about 20 min, then compare a matched section processed without retrieval to identify retrieval-dependent signal (standard IHC practice). If staining remains weak, adjust heating time on serial sections while holding antibody concentration and detection constant; excessive heating can damage morphology and increase background (standard IHC practice). Judge the result against the peptide-blocked prostate carcinoma image for SKU A30552, while recognizing that its caption does not report retrieval conditions (selected A30552 caption).
Could fixation explain weak DLC1 staining in my paraffin sections?
DLC1-specific sensitivity to fixation is unknown because the supplied tissue-IHC caption reports paraffin embedding but no fixative (selected A30552 caption). Record the fixative and exposure time for each specimen, then compare similarly processed sections before attributing weaker staining to biology (standard IHC practice). If using 10% neutral buffered formalin, keep fixation and subsequent processing consistent across comparison groups, and inspect morphology for underfixation or overprocessing (standard IHC practice). Neither the HPA tissue pattern nor DLC1 topology and phosphorylation sites establish a fixation effect for this antibody (HPA tissue IHC: reliability uncertain; UniProt Q96QB1 record).
Which staining compartments are plausible for DLC1, and how should I assess luminal signal?
Expect predominantly cytoplasmic signal, with possible enrichment at cell junctions or focal adhesions and the cell periphery (UniProt Q96QB1 localisation). DLC1 has no transmembrane segment, so a continuous membrane outline alone does not establish membrane insertion (UniProt Q96QB1 topology). HPA reports membranous and luminal tissue staining, but rates its tissue-IHC evidence Uncertain because staining and RNA expression show low consistency (HPA tissue IHC). Score cytoplasmic, peripheral and luminal staining separately in at least 2 matched sections, and confirm that compartment calls survive review of cell boundaries and background controls (standard IHC practice).
How could DLC1 isoforms or epitope placement change my IHC result?
DLC1 has 6 listed isoforms and SAM, Rho-GAP and START domains at residues 448–515, 1078–1284 and 1314–1521, respectively (UniProt Q96QB1 record). Its listed phosphoserines include residues 523, 526, 566 and 757, but the payload does not place the catalog antibody epitope relative to them (UniProt Q96QB1 record; selected A30552 caption). Request the immunogen sequence or mapped epitope before interpreting absent staining as loss of all DLC1 isoforms (standard IHC practice). The peptide-blocked A30552 tissue image supports competition by its synthesized peptide, but cannot by itself establish isoform coverage or distinguish specific tissue signal from every cross-reactivity concern (selected A30552 caption; standard IHC practice).
How should I design a companion IF experiment to clarify DLC1 IHC staining?
Use an adjacent matched section for IF and multiplex DLC1 with a glandular-cell marker, such as pan-cytokeratin, when examining HPA-reported glandular staining (HPA tissue IHC; standard IF practice). Choose a red or far-red DLC1 fluorophore after measuring unstained-section autofluorescence, and keep single-label controls for spectral bleed-through (standard IF practice). Because DLC1 is mainly cytosolic and has no transmembrane segment, test mild permeabilisation, such as 0.1% Triton X-100 for 5 min, against an unpermeabilised section if the epitope is accessible only inside cells (HPA subcellular; UniProt Q96QB1 topology; standard IF practice). Compare the resulting cell type and compartment with chromogenic IHC, without treating the paraffin tissue caption as evidence of IF validation (selected A30552 caption).
What should I check when DLC1 DAB staining appears diffuse or excessive?
Run a no-primary control and inspect whether color persists in the same structures, which would implicate detection chemistry or endogenous activity rather than primary-antibody binding (standard IHC practice). For peroxidase-based DAB detection, test a 3% hydrogen peroxide block for 10 min and a 5% normal-serum protein block for 20 min, adjusting them to the detection system (standard IHC practice). Titrate the primary antibody on matched sections and shorten DAB development if signal obscures cell boundaries (standard IHC practice). Compare the result with the peptide-blocked A30552 prostate carcinoma image, while remembering that peptide competition does not identify every possible source of background (selected A30552 caption; standard IHC practice).
How should I quantify DLC1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record the percentage of positive viable cells and intensity on a 0–3 scale (standard IHC practice). An H-score can summarize these observations from 0–300 by adding each intensity grade multiplied by its percentage of cells (standard IHC practice). Normalize cell-based scores to the number of viable cells examined, or report stained area per mm² of viable tissue when cell counting is impractical (standard IHC practice). Keep cytoplasmic and luminal measurements separate because HPA reports luminal staining with Uncertain tissue-IHC reliability, and use matched processing and thresholds across groups (HPA tissue IHC; standard IHC practice).
How can I distinguish plausible DLC1 signal from staining artefact?
Treat cytoplasmic staining with coherent cell-peripheral or focal-adhesion enrichment as biologically plausible for DLC1 (UniProt Q96QB1 localisation). Check whether stained cells match the tissue compartment under review; HPA reports glandular-cell signal in gallbladder and colon but rates its overall tissue-IHC reliability Uncertain (HPA tissue IHC). Be skeptical of nuclear-only staining, section-edge gradients, necrotic areas or color retained in a no-primary control, and check the peroxidase block when endogenous enzyme activity is suspected (UniProt Q96QB1 localisation; standard IHC practice). The peptide-blocked A30552 prostate carcinoma image is a useful comparison, but interpretation still requires morphology, matched controls and reproducibility across sections (selected A30552 caption; standard IHC practice).
Boster reagents
Best DLC1 / Rho GTPase-activating protein 7 IHC Antibodies
A30552 has IHC data from paraffin-embedded human prostate carcinoma tissue and IF data from A549 cells (catalog image captions). Human, Mouse, and Rat reactivity is listed (catalog reactivity).
Real IHC data
Anti-RHG07 DLC1 Antibody
Cat # A30552
A30552 shows IHC staining of paraffin-embedded human prostate carcinoma tissue (catalog IHC image caption). It also shows IF staining of A549 cells and is listed for IHC, IF, and ICC (catalog IF image caption; catalog applications).
Which to pick: Choose A30552 for tissue IHC: its IHC image uses paraffin-embedded human prostate carcinoma tissue, but the fixative is unreported (catalog IHC image caption). Choose A30552 for IF/ICC because those applications are listed and its IF image shows A549 cells (catalog applications; catalog IF image caption). For cross-species work, A30552 is a rabbit polyclonal antibody with Human, Mouse, and Rat reactivity listed; the supplied IHC image documents human tissue only (catalog host; catalog dilution_raw; catalog reactivity; catalog IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96QB1 (RHG07_HUMAN, Rho GTPase-activating protein 7).
- Human Protein Atlas. DLC1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. DLC1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the endoplasmic reticulum and vesicles..
- Human Protein Atlas. DLC1 antibody validation summary (3 antibodies).
- Low expression of DLC1 is predictive of poor therapeutic efficiency of fluoropyrimidine and oxaliplatin as adjuvant chemotherapy in gastric cancer. Molecular medicine reports 2015 — PMC4581752.
- Aberrant DNA methylation of alternative promoter of DLC1 isoform 1 in meningiomas. Journal of neuro-oncology 2016 — PMC5118400.
- Curcumin inhibits the growth of triple-negative breast cancer cells by silencing EZH2 and restoring DLC1 expression. Journal of cellular and molecular medicine 2020 — PMC7521266.
- Nuclear-targeted deleted in liver cancer 1 (DLC1) is less efficient in exerting its tumor suppressive activity both in vitro and in vivo. PloS one 2011 — PMC3180446.
- PubMed PMID:9605766 — UniProt-cited evidence.
- PubMed PMID:11214970 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DLC1 (Rho GTPase-activating protein 7). Expected localisation Membranous and luminal tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A30552 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Gallbladder (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate carcinoma tissue; fixative not specified (datasheet A30552); 2. Fixation: Image fixative and duration unreported (datasheet A30552); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-DLC1, 1:100 - 1:300 (datasheet A30552); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Staining has low agreement with RNA; verification is pending (HPA tissue IHC); Isoform / epitope: 6 isoforms; antibody epitope coverage is unknown (UniProt); Regulation: Expression regulation is not established (UniProt). Evidence: UniProt Q96QB1, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A30552, PMC4581752, PMC5118400. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DLC1/immunohistochemistry-ihc-guide.