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GNL3L / Guanine nucleotide-binding protein-like 3-like protein · IHC design guide
Design Immunohistochemistry for GNL3L
Plan paraffin-section GNL3L IHC with the catalog antibody’s 1:100–1:300 dilution range (datasheet A09415). Assess nuclear and cytoplasmic staining across cell types, with high staining in adrenal gland glandular cells and no detected staining in lymph-node germinal-center cells as tissue references (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GNL3L 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 | Nuclear and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Many cell types show cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A09415) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Lymph node |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Germinal-center cells are undetected despite broad staining (HPA tissue IHC) | |
| Regulation | No expression regulation annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms or cleavage to shift epitopes (UniProt) |
Section 1
Recommended GNL3L IHC & IF Protocols
The catalog antibody protocol (datasheet A09415) is followed by three published GNL3L IHC protocols using human esophageal specimens (PMC12378702; PMC11087641; PMC8154413).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A09415) |
| Fixation | Image fixative and duration unreported (datasheet A09415); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A09415); 20 min, 95–100 °C (standard) |
| 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-GNL3L, 1:100-1:300 (datasheet A09415) |
| 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 | GNL3L-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA, pH 9.0 (datasheet A09415). Citric acid retrieval is a published alternative, though its pH and heating conditions were not specified (PMC11087641).
Section 2
What Is the Expected GNL3L Staining Pattern?
GNL3L is a nuclear and nucleolar protein with no transmembrane segment (UniProt Q9NVN8: localization and topology). Tissue IHC shows widespread nuclear and cytoplasmic staining, with high staining in several glandular, squamous epithelial, and neural cell populations (HPA tissue IHC). HPA rates the tissue pattern Supported, with medium agreement between staining and RNA data and external verification pending (HPA tissue IHC).
| Nuclear staining, sometimes with cytoplasmic staining, in adrenal glandular or cervical squamous epithelial cells. | This fits the reported tissue pattern; both cell populations have High staining (HPA tissue IHC). A nucleolar accent would also agree with the subcellular reference, which comes from ICC-IF rather than tissue IHC (HPA subcellular ICC-IF). |
| Predominantly cell-surface staining, with little nuclear signal. | Treat a membrane-dominant pattern as suspect: GNL3L has no transmembrane segment and is assigned to the nucleus and nucleolus (UniProt Q9NVN8: topology and localization). Recheck the signal against a known-positive tissue before assigning it to GNL3L (general IHC practice). |
| Strong staining in lymph-node germinal center cells. | That conflicts with the reported Not detected result for these cells (HPA tissue IHC). Consider cross-reactivity or endogenous chromogenic activity, and assess control sections; the HPA result is a reference pattern, not proof that every germinal center must be negative (HPA tissue IHC; general IHC practice). |
| Uniform chromogen across cells and surrounding tissue, with no clear cellular boundaries. | Diffuse background does not support a compartment assignment (general IHC practice). Check blocking, washes, and the detection-only control; interpret any remaining signal against the nuclear and cytoplasmic tissue pattern (HPA tissue IHC). |
| No signal in adrenal glandular cells or cervical squamous epithelial cells. | Both are reported High examples, so an absent signal calls for a technical check (HPA tissue IHC). Review tissue integrity, antibody application, retrieval conditions, and detection controls using the chosen assay workflow (general IHC practice). |
💡Expected GNL3L appearanceCall a result positive when identifiable cells show nuclear staining, with possible cytoplasmic staining, including High signal in an HPA High reference population; isolated membrane-dominant signal or diffuse chromogen without cellular definition is suspect (HPA tissue IHC; UniProt Q9NVN8: topology; general IHC practice).
| Tissue and cell population | Intensity depends on the cells being assessed: adrenal glandular and cervical squamous epithelial cells are High; endometrial glandular cells are Medium; lung alveolar type I cells are Low; lymph-node germinal center cells are Not detected (HPA tissue IHC). Score the named population rather than the whole section (general IHC practice). |
| Compartment reference | Tissue IHC is described as ubiquitously cytoplasmic and nuclear (HPA tissue IHC). ICC-IF places GNL3L mainly in nucleoli, with additional nucleoplasm and cytosol (HPA subcellular ICC-IF). Use that finer location as supporting context, while allowing the tissue IHC pattern to guide slide interpretation. |
| Antibody evidence | The two listed antibodies, HPA036314 and HPA036315, are each Supported for IHC; HPA036315 is also Supported for ICC (HPA antibodies). The tissue-level Supported rating has medium staining–RNA consistency and awaits external verification (HPA tissue IHC). Neither designation guarantees specificity in an individual section. |
| Topology and processing | The record gives a 582-aa chain, no signal peptide or propeptide, no transmembrane segment, and no annotated glycosylation sites or isoforms (UniProt Q9NVN8). These features provide no basis to predict a membrane or secreted staining pattern; they do not establish epitope accessibility after fixation. |
| IF/ICC Q&A | Q: What pattern should IF/ICC show? A: Mainly nucleoli, with additional nucleoplasmic and cytosolic signal (HPA subcellular ICC-IF). HPA lists images from A-431, U-251MG, and U2OS and rates HPA036315 Supported for ICC (HPA subcellular ICC-IF; HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| A High reference population has no chromogenic signal. | The result disagrees with the reported High staining in adrenal glandular or cervical squamous epithelial cells (HPA tissue IHC); the cause cannot be identified from the pattern alone. | Confirm tissue identity and integrity, then inspect retrieval, primary-antibody application, and detection controls within the chosen IHC workflow (general IHC practice). |
| The slide shows cytoplasmic staining but little obvious nucleolar detail. | Tissue IHC reports nuclear and cytoplasmic expression, while the mainly nucleolar assignment comes from ICC-IF (HPA tissue IHC; HPA subcellular ICC-IF). The two assays need not resolve compartments equally (general microscopy practice). | Judge whether nuclear staining and the expected cell distribution are present; use the ICC-IF reference for finer localization without requiring visible nucleoli in every paraffin section (HPA tissue IHC; HPA subcellular ICC-IF). |
| Chromogen appears concentrated at cell borders. | A membrane-dominant pattern conflicts with the nuclear/nucleolar assignment and lack of a transmembrane segment (UniProt Q9NVN8: localization and topology). | Compare a known-positive population and control sections, and check whether the apparent border signal follows tissue edges or extends beyond cells (HPA tissue IHC; general IHC practice). |
| Diffuse brown staining obscures cell boundaries. | Nonspecific binding or endogenous detection activity can produce background in chromogenic IHC (general IHC practice); appearance alone cannot identify which applies. | Inspect a detection-only control, review blocking and washes, and score only signal that remains clearly cellular and compartmentalized (general IHC practice). |
| Germinal center cells stain strongly while nearby cells also look positive. | Germinal center cells are reported Not detected, despite the broader ubiquitous tissue profile (HPA tissue IHC). Strong staining there may reflect cross-reactivity or endogenous activity (general IHC practice). | Identify the stained cell population on the counterstained section and compare control sections before treating the germinal center signal as target-specific (general IHC practice). |
| Signal intensity differs between sections of the same reference tissue. | The HPA tissue profile describes cell populations, and its Supported rating has medium agreement with RNA data; it does not establish a target-specific fixation effect (HPA tissue IHC). Section or workflow differences require separate assessment (general IHC practice). | Compare the same cell population across sections and review section quality, retrieval, and detection records before assigning a biological intensity difference (general IHC practice). |
Experiment evidence
IHC & IF images showing GNL3L staining
A09415 · IHCImmunohistochemical analysis of paraffin-embedded human brain. 1, Tris-EDTA, pH9.0 was used for antigen retrieval. 2 Antibody was diluted at
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000130119-GNL3L/tissue/cerebral+cortex#imid_9721492
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000130119-GNL3L/tissue/colon#imid_9721410
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000130119-GNL3L/tissue/liver#imid_9721403
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000130119-GNL3L/tissue/kidney#imid_9721404
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036315; green = GNL3L, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036315; green = GNL3L, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GNL3L IHC & IF
🧪Run adrenal gland first; its glandular cells should show GNL3L staining (HPA: High in adrenal gland glandular cells). Run lymph node as the negative tissue, focusing on germinal center cells (HPA: Not detected in lymph node germinal center cells); on the positive slide, cells without specific nucleolar signal should show only background, but the supplied HPA rows do not identify a validated internal negative cell type (HPA: adrenal gland glandular cells High).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Lymph node (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GNL3L in A-431, U-251MG, U2OS, with annotated localisation: Nucleoli (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and antibody-class-matched isotype controls, plus a GNL3L knockout sample or immunizing-peptide competition if available. For chromogenic IHC in adrenal gland, block endogenous peroxidase and check the no-primary slide for tissue pigment or other background.
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected A09415 paraffin-section brain caption does not state a fixative (A09415 tissue-IHC caption: fixative unreported). Tris-EDTA at pH 9.0 is a documented retrieval starting point, but retrieval dependence is untested in the supplied evidence (A09415 tissue-IHC caption: Tris-EDTA pH 9.0). Is frozen-section IF easier? The supplied evidence does not establish that comparison; ICC-IF images support nucleolar localization, with additional nucleoplasmic and cytosolic localization (HPA subcellular: nucleoli supported; nucleoplasm and cytosol approved), and no tissue-specific artefact is documented.
HPA tissue IHC evidence for GNL3L
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GNL3L IHC Tips
Troubleshoot GNL3L staining by checking retrieval, controls, and cellular compartment before comparing signal across paraffin sections (datasheet A09415; HPA subcellular).
How should I retrieve GNL3L antigen when nucleolar staining is weak?
Use heat-mediated Tris-EDTA at pH 9.0 for antigen retrieval in paraffin sections (datasheet A09415). The selected human brain image used this retrieval with primary antibody at 1:200 overnight at 4 °C (caption A09415). Keep heating and cooling consistent across slides, and compare adjacent sections with a no-primary control (standard IHC practice). If staining remains weak, adjust retrieval duration incrementally while checking morphology and nonspecific nuclear color (standard IHC practice). Judge improvement by clearer nucleolar enrichment, allowing some nucleoplasmic signal, rather than by overall darkness (HPA subcellular; standard IHC practice).
Can fixation explain variable GNL3L staining across paraffin blocks?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state a fixative (caption A09415). Record each block's fixative, fixation duration, processing history, and section age before comparing staining (standard IHC practice). Compare similarly processed sections using the same Tris-EDTA pH 9.0 retrieval and antibody dilution of 1:200 (datasheet A09415; caption A09415). If signal varies, examine morphology, retrieval consistency, and control-slide staining before assigning a cause (standard IHC practice). HPA tissue patterns and GNL3L's intracellular location cannot establish how this antigen responds to fixation (HPA tissue IHC; UniProt Q9NVN8; standard IHC interpretation).
Should cytoplasmic staining count as a GNL3L-positive result?
Prioritize distinct nucleolar staining when evaluating GNL3L in chromogenic sections (UniProt Q9NVN8 subcellular; HPA subcellular). Nucleoplasmic and cytosolic signal can also occur, while the tissue IHC profile describes widespread nuclear and cytoplasmic expression (HPA subcellular; HPA tissue IHC). Score these compartments separately instead of treating all brown staining as equivalent target signal (standard IHC practice). Check whether signal sits within intact cells and whether a no-primary slide shows similar cytoplasmic color (standard IHC practice). Diffuse staining without convincing cellular boundaries or nucleolar enrichment needs cautious interpretation, especially near damaged tissue (HPA subcellular; standard IHC practice).
How do I assess unexpected GNL3L staining without an epitope map?
The supplied record lists a single 1–582 amino-acid chain and 0 annotated isoforms for GNL3L (UniProt Q9NVN8). It also lists a CP-type G domain at residues 125–310, with no transmembrane segment or annotated glycosylation sites (UniProt Q9NVN8). Those annotations do not identify the catalog antibody's epitope or show that retrieval exposes every region equally (UniProt Q9NVN8; standard IHC interpretation). Check the antibody's stated immunogen or epitope information before proposing a domain-specific explanation (standard IHC practice). Meanwhile, compare nucleolar localisation and control-slide staining under the documented pH 9.0 retrieval (HPA subcellular; datasheet A09415; standard IHC practice).
How should I cross-check GNL3L localisation with multiplex IF?
Use IF as a separate localisation check, pairing GNL3L with a validated marker for the cell type under study (standard IF practice). For cerebral cortex, a glial-cell marker is relevant because HPA reports high GNL3L staining in glial cells (HPA tissue IHC). Choose spectrally separated fluorophores, assigning the dimmer channel a wavelength with less measured tissue autofluorescence (standard IF practice). GNL3L lacks a transmembrane segment and is mainly nucleolar, so permeabilisation must permit antibody access to intracellular nuclear antigen; the antibody's exact epitope is unspecified (UniProt Q9NVN8; HPA subcellular). Include single-color and no-primary controls before comparing IF localisation with chromogenic IHC (standard IF practice).
What should I check when brown precipitate obscures GNL3L localisation?
Inspect a no-primary section first to distinguish detection-system color from antibody-dependent staining (standard IHC practice). Include a peroxidase block before chromogenic detection and assess endogenous pigment or precipitate in the same tissue region (standard IHC practice). The selected image used primary antibody at 1:200 overnight at 4 °C and secondary antibody at 1:200 for 45 minutes at room temperature (caption A09415). If background persists, titrate primary antibody and shorten chromogen development while preserving the controls (standard IHC practice). Accept improvement only when cellular detail remains clear and expected nucleolar staining is retained (HPA subcellular; standard IHC practice).
How should I score heterogeneous GNL3L staining in tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and eligible cell population before scoring, excluding folds, damaged edges, and necrosis (standard IHC practice). Record the percentage of positive cells and staining intensity by compartment; an H-score can summarize intensity across cells (standard IHC practice). Report nucleolar and nucleoplasmic scores separately because GNL3L is mainly nucleolar but also occurs in the nucleoplasm (HPA subcellular). Normalize positive-cell counts to all eligible cells in the sampled region, or report positive-cell density per mm² of evaluable tissue (standard IHC practice). Keep retrieval, antibody dilution, detection, counterstain, and image thresholds consistent across compared sections (datasheet A09415; standard IHC practice).
How can I distinguish convincing GNL3L positivity from staining artefact?
Look for staining within intact cells, with nucleolar enrichment as the strongest localisation clue for GNL3L (HPA subcellular; standard IHC practice). HPA also reports nucleoplasmic and cytosolic localisation, so these compartments require comparison with cellular morphology and controls (HPA subcellular; standard IHC practice). Cell identity matters: HPA reports high staining in cerebral-cortex glial cells but no detected staining in lymph-node germinal-center cells (HPA tissue IHC). Discount color concentrated at section edges or necrotic areas, and investigate staining reproduced on a no-primary control for endogenous enzyme activity (standard IHC practice). Treat tissue comparisons cautiously because HPA rates its tissue IHC profile Supported, with medium agreement to RNA data and external verification pending (HPA tissue IHC).
Boster reagents
Best GNL3L / Guanine nucleotide-binding protein-like 3-like protein IHC Antibodies
A09415 has an IHC image from paraffin-embedded human brain (IHC caption). IF is listed without an image, and Human, Mouse, Rat and Cat reactivity is listed (catalog applications/reactivity/IF images).
Real IHC data
Anti-GNL3L Antibody
Cat # A09415
A09415 will render with its IHC figure from paraffin-embedded human brain, showing Tris-EDTA pH 9.0 retrieval and primary antibody at 1:200 overnight at 4°C (A09415 IHC caption). Its catalog lists IHC and IF applications and Human, Mouse, Rat and Cat reactivity; no IF image is supplied (A09415 catalog applications/reactivity/IF images).
Which to pick: For paraffin-section chromogenic IHC, pick A09415 based on its own human brain IHC figure; the fixative is unreported (A09415 IHC caption). For IF, A09415 is listed at 1:50, while ICC and IF imaging are undocumented (A09415 catalog applications/dilution/IF images). For cross-species work, A09415 lists Human, Mouse, Rat and Cat reactivity, although its IHC figure documents only human brain; clonality is unreported (A09415 catalog reactivity/clone; IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NVN8 (GNL3L_HUMAN, Guanine nucleotide-binding protein-like 3-like protein).
- Human Protein Atlas. GNL3L tissue IHC expression (reliability: Supported).
- Human Protein Atlas. GNL3L subcellular location (ICC-IF): Mainly localized to the nucleoli. In addition localized to the nucleoplasm and cytosol..
- Human Protein Atlas. GNL3L antibody validation summary (2 antibodies).
- The GNL3L-MDM2 Interaction Drives Esophageal Squamous Cell Carcinoma Progression. Cancer medicine 2025 — PMC12378702.
- Multi-Omics Analysis of GNL3L Expression, Prognosis, and Immune Value in Pan-Cancer. Cancers 2022 — PMC9558978.
- Revealing the oncogenic role of elevated GNL3L expression in esophageal squamous cell carcinoma: insights into the STAT3 pathway. Journal of thoracic disease 2024 — PMC11087641.
- High expression of guanine nucleotide-binding protein-like-3-like is associated with poor prognosis in esophageal cancer. Medicine 2021 — PMC8154413.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15772651 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GNL3L (Guanine nucleotide-binding protein-like 3-like protein). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A09415). Validated IHC antibody: A09415 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Lymph node. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A09415); 2. Fixation: Image fixative and duration unreported (datasheet A09415); 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: Tris-EDTA pH 9.0 (datasheet A09415); 20 min, 95–100 °C (standard); 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-GNL3L, 1:100-1:300 (datasheet A09415); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Germinal-center cells are undetected despite broad staining (HPA tissue IHC); Isoform / epitope: No annotated isoforms or cleavage to shift epitopes (UniProt); Regulation: No expression regulation annotated (UniProt). Evidence: UniProt Q9NVN8, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoli. In addition localized to the…, Datasheet A09415, PMC12378702, PMC11087641. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GNL3L/immunohistochemistry-ihc-guide.