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L3MBTL3 / Lethal(3)malignant brain tumor-like protein 3 · IHC design guide
Design Immunohistochemistry for L3MBTL3
Plan L3MBTL3 paraffin-section IHC around the reported cytoplasmic tissue staining (HPA tissue IHC) and annotated nuclear location (UniProt). Start with the catalog antibody’s 2–5 μg/ml IHC range (datasheet A10834-1), and include specificity controls because staining and RNA have low concordance (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
L3MBTL3 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic in tissue IHC (HPA tissue IHC); nuclear location (UniProt) | |
| Staining pattern | General cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A10834-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10834-1) | |
| Caveat | Staining has low concordance with RNA; verify specificity (HPA tissue IHC) | |
| Regulation | Low tissue specificity at RNA level (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; check epitope coverage (UniProt) |
Section 1
Recommended L3MBTL3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A10834-1) with the published gastric tissue IHC protocol (PMC10778146).
| Sample | Paraffin-embedded human bladder urothelial carcinoma tissue; fixative not specified (datasheet A10834-1) |
| Fixation | Image fixative and duration unreported (datasheet A10834-1); 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 A10834-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A10834-1) |
| Primary antibody | Rabbit anti-L3MBTL3, 2-5 μg/ml (datasheet A10834-1) |
| Primary incubation | Overnight at 4 °C (datasheet A10834-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A10834-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | L3MBTL3-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: General cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet: A10834-1); consider sodium citrate at 95 °C when adapting the published gastric tissue protocol (PMC10778146).
Section 2
What Is the Expected L3MBTL3 Staining Pattern?
L3MBTL3 is a nuclear, non-membrane protein (UniProt Q96JM7: nucleus; no transmembrane segment). For paraffin-section IHC, HPA reports general cytoplasmic staining, including medium staining in kidney tubular cells and bronchial respiratory epithelial cells (HPA tissue IHC). HPA rates its tissue IHC Approved but reports low agreement with RNA expression and pending external verification (HPA tissue IHC). Interpret the cytoplasmic pattern with that limitation in view.
| Kidney tubular cells or bronchial respiratory epithelial cells show moderate cytoplasmic chromogen, with identifiable cell boundaries. | This matches the reported medium IHC staining in those cells (HPA tissue IHC). Score the cell type, compartment and intensity together; the observation alone does not resolve the difference from UniProt's nuclear localization (UniProt Q96JM7: nucleus). |
| Signal is confined to cell membranes, extracellular material or tissue edges, without convincing cellular staining. | That compartment does not match either the reported cytoplasmic tissue IHC profile (HPA tissue IHC) or the nuclear localization (UniProt Q96JM7). Treat it as a possible staining artefact and check its distribution against a matched negative control (general IHC practice). |
| Strong signal appears in adipocytes or colon glandular cells while the expected cell population is weak. | HPA reports these cell populations as not detected (HPA tissue IHC). Consider antibody cross-reactivity or endogenous detection activity, particularly if the signal persists in a no-primary control; HPA's Approved rating does not verify every tissue pattern (HPA tissue IHC; general IHC practice). |
| Chromogen is diffuse across the section or fills multiple tissue compartments without clear cell outlines. | This is hard to score as cell-specific L3MBTL3 staining (general IHC practice). Compare a no-primary control and inspect the counterstain, section edges and pigment before assigning a compartment; HPA's cytoplasmic profile describes cellular staining, not a uniform haze (HPA tissue IHC). |
| Kidney tubular cells show no detectable signal in a run expected to show a positive reference pattern. | HPA reports medium staining in kidney tubular cells, making them a useful reference for this comparison (HPA tissue IHC). A negative result calls for review of section quality, retrieval and detection controls; it does not establish absence of L3MBTL3, given the reported IHC/RNA discordance (HPA tissue IHC; general IHC practice). |
💡Expected L3MBTL3 appearanceFor IHC, a plausible positive is clearly cellular, medium cytoplasmic staining in kidney tubular or bronchial respiratory epithelial cells (HPA tissue IHC); edge-only, extracellular or uniform diffuse colour is suspect (general IHC practice).
| Compartment evidence | UniProt assigns L3MBTL3 to the nucleus, while HPA describes general cytoplasmic tissue IHC (UniProt Q96JM7; HPA tissue IHC). HPA ICC-IF supports nucleoplasmic and nucleolar localization (HPA subcellular). Report the observed IHC compartment explicitly instead of treating either source as confirmation of the other. |
| IHC antibody validation | HPA053035 has Approved IHC status; HPA044382 has no listed IHC status (HPA antibodies). The HPA tissue profile also has low agreement with RNA expression and awaits external verification (HPA tissue IHC). An IHC result that drives a conclusion needs independent support, such as a second validated reagent or an appropriate control (general IHC practice). |
| Protein features | UniProt lists 2 isoforms, a single chain spanning residues 1–780, no signal peptide, no propeptide and no transmembrane segment (UniProt Q96JM7). These annotations provide no basis for predicting a secreted or membrane pattern; the supplied record does not identify the catalog antibody's epitope or establish isoform-specific staining. |
| IF/ICC question: where should signal appear? | Nucleoplasm and nucleoli are supported main locations in HPA ICC-IF images; vesicles, the cytokinetic bridge and primary cilium are uncertain additional locations (HPA subcellular). Both listed HPA antibodies have Enhanced ICC validation, while only HPA053035 has listed IHC approval (HPA antibodies). This answers localization for IF/ICC without extending its protocol to paraffin IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in kidney tubular cells. | The reference population is reported at medium intensity (HPA tissue IHC); a failed detection run or weak assay conditions are possibilities (general IHC practice). | Confirm tissue and cellular preservation, then review the run's positive control, retrieval settings, antibody dilution and detection reagents (general IHC practice). Do not infer target absence from one negative section. |
| Broad staining remains in a no-primary control. | Primary-independent background, including endogenous detection activity, is possible (general IHC practice). | Review the chromogenic detection system and its appropriate endogenous-activity block; compare the control with the test section before scoring L3MBTL3 (general IHC practice). |
| Strong staining is limited to adipocytes or colon glandular cells. | Those populations are listed as not detected (HPA tissue IHC); cross-reactivity or background may explain a discordant result (general IHC practice). | Check a no-primary control and compare cell-specific staining in a reported medium-staining tissue on the same run (HPA tissue IHC; general IHC practice). |
| Signal appears only at section edges or in extracellular deposits. | Neither location matches the reported cellular IHC profile or nuclear annotation (HPA tissue IHC; UniProt Q96JM7). Edge-associated stain may reflect slide artefact (general IHC practice). | Inspect adjacent tissue, section integrity and the no-primary control; exclude edge-only deposits from cell-level scoring (general IHC practice). |
| IHC is cytoplasmic, but IF/ICC images are nuclear. | The sources themselves differ: HPA tissue IHC reports general cytoplasmic staining; HPA ICC-IF supports nucleoplasm and nucleoli, consistent with UniProt's nuclear annotation (HPA tissue IHC; HPA subcellular; UniProt Q96JM7). | Record assay and compartment separately. Reassess IHC specificity with controls and independent evidence before calling cytoplasmic staining definitive L3MBTL3 localization (general IHC practice). |
| Signal is weak or uneven across a section. | Uneven staining can arise from section or reagent handling (general IHC practice); the supplied sources do not establish L3MBTL3-specific fixation sensitivity. | Compare tissue morphology and the run's controls, then check retrieval, reagent coverage and detection consistency (general IHC practice). Interpret remaining differences against cell-specific HPA observations rather than section-wide colour alone (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing L3MBTL3 staining
A10834-1 · IHCIHC analysis of L3MBTL3 using anti-L3MBTL3 antibody (A10834-1). L3MBTL3 was detected in a paraffin-embedded section of human bladder urothel
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000198945-L3MBTL3/tissue/cerebral+cortex#imid_14750084
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000198945-L3MBTL3/tissue/colon#imid_14772313
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000198945-L3MBTL3/tissue/liver#imid_14772302
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000198945-L3MBTL3/tissue/kidney#imid_14772264
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA044382; green = L3MBTL3, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA044382; green = L3MBTL3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for L3MBTL3 IHC & IF
🧪Run adrenal gland first and score its glandular cells for nuclear staining (HPA: Medium in adrenal gland glandular cells; UniProt: nucleus). Run adipose tissue as the negative comparison (HPA: Not detected in adipocytes); on the adrenal slide, non-glandular cells can serve as candidate internal negatives if they lack nuclear chromogen, but their target-negative status requires confirmation (HPA: adrenal gland result specifies glandular cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show L3MBTL3 in HEK293, U2OS, CACO-2, KOLF2.1J, with annotated localisation: Nucleoplasm (supported), Nucleoli (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a rabbit IgG isotype control matched to the primary antibody’s clonality, and identically processed L3MBTL3-knockout material as a biological negative (selected-SKU caption: rabbit primary and peroxidase-conjugated anti-rabbit secondary). Quench endogenous peroxidase and check the adrenal section for pigment that could be mistaken for DAB signal (selected-SKU caption: HRP/DAB detection; HPA: adrenal gland positive tissue).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU tissue-IHC caption does not state a fixative (selected-SKU caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used for the captioned paraffin-section result, but a retrieval requirement for adrenal gland is unreported (selected-SKU caption: EDTA retrieval in a paraffin section of bladder urothelial carcinoma). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; for adrenal IHC, assess pigment separately from nuclear DAB staining (HPA: adrenal gland positive tissue; UniProt: nucleus).
HPA tissue IHC evidence for L3MBTL3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Colon | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced L3MBTL3 IHC Tips
Troubleshoot L3MBTL3 staining in paraffin sections by comparing nuclear signal, tissue morphology, and matched controls (UniProt Q96JM7; HPA subcellular).
Which retrieval condition should I try first for L3MBTL3 IHC?
Begin with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A10834-1). Keep heating and cooling conditions consistent across paired sections, then adjust heating duration if signal is weak or morphology suffers (standard IHC practice). The documented starting condition uses 2 μg/ml primary antibody overnight at 4°C (datasheet A10834-1). Assess nuclear staining because L3MBTL3 is nuclear, with supported nucleoplasmic and nucleolar locations (UniProt Q96JM7; HPA subcellular). If staining remains weak, compare another retrieval condition against the EDTA baseline while holding antibody concentration and detection constant (standard IHC practice).
Could fixation explain weak or uneven L3MBTL3 staining?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state a fixative (datasheet A10834-1). Record each specimen’s fixative and fixation duration before comparing staining intensity across cases (standard IHC practice). For newly prepared material, evaluate a documented 10% neutral buffered formalin condition as a general IHC starting point, without assuming it preserves this epitope optimally (standard IHC practice). Compare matched sections with identical EDTA retrieval at pH 8.0 and identical detection settings (datasheet A10834-1; standard IHC practice). If morphology and signal vary together, assess tissue processing before attributing the difference to L3MBTL3 abundance (standard IHC practice).
How should I assess cytoplasmic versus nuclear L3MBTL3 staining?
Inspect nuclei first: UniProt assigns L3MBTL3 to the nucleus, and HPA supports nucleoplasmic and nucleolar localization (UniProt Q96JM7; HPA subcellular). Record nuclear and cytoplasmic staining separately rather than combining them into one score (standard IHC practice). HPA tissue IHC describes general cytoplasmic expression, but its Approved entry reports low agreement with RNA data and awaits external verification (HPA tissue IHC). If cytoplasmic DAB dominates, examine matched negative controls and repeat staining with EDTA retrieval at pH 8.0 (datasheet A10834-1; standard IHC practice). Treat vesicle, cytokinetic bridge, or cilium assignments cautiously because HPA labels them uncertain (HPA subcellular).
Could isoforms or epitope masking change my IHC result?
L3MBTL3 has 2 annotated isoforms, so interpretation depends on which sequence the antibody recognizes (UniProt Q96JM7). Check the antibody’s stated immunogen or epitope against both isoforms before interpreting a negative section as absence of all L3MBTL3 (standard IHC practice; UniProt Q96JM7). The protein has a SAM domain at residues 708–772 and an annotated phosphoserine at 608, but the supplied caption does not map the antibody epitope (UniProt Q96JM7; datasheet A10834-1). Compare paired sections under the documented EDTA pH 8.0 retrieval condition (datasheet A10834-1). Avoid assigning an isoform-specific staining pattern without epitope mapping or an independent specificity control (standard IHC practice).
How can IF help investigate an ambiguous IHC pattern?
Use IF as a localization cross-check while interpreting the chromogenic section, because HPA supports nucleoplasmic and nucleolar signal in ICC/IF images (HPA subcellular). In the documented urothelial carcinoma specimen, multiplex L3MBTL3 with an epithelial marker to identify the stained cell population (datasheet A10834-1; standard IF practice). Choose a fluorophore channel after checking tissue autofluorescence with an unstained control (standard IF practice). Apply permeabilisation appropriate for a nuclear epitope and compare it with a matched control, since L3MBTL3 has no transmembrane segment (UniProt Q96JM7; standard IF practice). Keep IF conclusions separate from the catalog antibody’s paraffin-section performance (datasheet A10834-1; standard IHC practice).
What should I check when L3MBTL3 DAB staining is diffuse?
Run a no-primary control to identify signal from secondary reagents, endogenous peroxidase, or the chromogen (standard IHC practice). Apply a peroxidase block before HRP/DAB detection and compare background with the documented 10% goat serum block (standard IHC practice; datasheet A10834-1). The caption uses 2 μg/ml primary antibody overnight at 4°C; titrate from that condition if diffuse staining persists (datasheet A10834-1; standard IHC practice). Check whether signal follows tissue edges, folds, or damaged areas rather than intact nuclei (standard IHC practice; UniProt Q96JM7). Interpret widespread cytoplasmic stain cautiously given the HPA tissue IHC reliability caveat (HPA tissue IHC).
How should I quantify L3MBTL3 in paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear L3MBTL3 separately from cytoplasmic DAB because supported localization is nucleoplasmic and nucleolar (HPA subcellular; standard IHC practice). Report the percentage of positive nuclei and an H-score from 0–300, using consistent intensity thresholds across slides (standard IHC practice). Normalize counts to evaluable nuclei or a defined tissue area, such as positive cells per mm² (standard IHC practice). Exclude necrosis, folds, and section edges using the same rules for every specimen (standard IHC practice). Record retrieval and staining batch alongside scores because the documented assay uses EDTA at pH 8.0 (datasheet A10834-1; standard IHC practice).
When does an L3MBTL3-positive result need further validation?
Give greatest weight to reproducible nucleoplasmic or nucleolar staining in intact cells, consistent with supported L3MBTL3 localization (HPA subcellular; standard IHC practice). Check that staining belongs to the intended cell population; the documented catalog image is from human bladder urothelial carcinoma tissue (datasheet A10834-1). Signal confined to edges, folds, or necrotic regions warrants review as a processing artefact (standard IHC practice). Diffuse DAB that persists without primary antibody suggests endogenous enzyme activity or detection background (standard IHC practice). Seek an independent specificity check before calling isolated cytoplasmic staining positive, since HPA tissue IHC reports low agreement with RNA expression and awaits external verification (HPA tissue IHC).
Boster reagents
Best L3MBTL3 / Lethal(3)malignant brain tumor-like protein 3 IHC Antibodies
One human-reactive anti-L3MBTL3 antibody has a real IHC image from a paraffin section of bladder urothelial carcinoma (A10834-1 catalog; A10834-1 image caption). No IF image is supplied (catalog IF images).
Real IHC data
Anti-L3MBTL3 Antibody ®
Cat # A10834-1
A10834-1 is listed for human IHC, with a figure showing staining in a paraffin section of human bladder urothelial carcinoma (catalog applications/reactivity; A10834-1 image caption). The catalog supplies no IF/ICC validation or figure for A10834-1 (catalog applications; catalog IF images).
Which to pick: Choose A10834-1 for human paraffin-section IHC; its listed concentration is 2–5 μg/ml, and its figure used 2 μg/ml after EDTA pH 8.0 heat retrieval (datasheet; A10834-1 image caption). The image caption does not report the fixative; A10834-1 is rabbit-hosted, with clonality unreported (A10834-1 image caption; catalog host/clone). No listed SKU has IF/ICC or nonhuman reactivity validation, so this payload supports no IF/ICC or cross-species recommendation (catalog applications/reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96JM7 (LMBL3_HUMAN, Lethal(3)malignant brain tumor-like protein 3).
- Human Protein Atlas. L3MBTL3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. L3MBTL3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nucleoli. In addition localized to vesicles, cytokinetic bridge and primary cilium..
- Human Protein Atlas. L3MBTL3 antibody validation summary (2 antibodies).
- L3MBTL3 Is a Potential Prognostic Biomarker and Correlates with Immune Infiltrations in Gastric Cancer. Cancers 2023 — PMC10778146.
- L3MBTL3 and STAT3 collaboratively upregulate SNAIL expression to promote metastasis in female breast cancer. Nature communications 2025 — PMC11696420.
- PubMed PMID:11347906 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for L3MBTL3 (Lethal(3)malignant brain tumor-like protein 3). Expected localisation Cytoplasmic in tissue IHC (HPA tissue IHC); nuclear location (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A10834-1). Validated IHC antibody: A10834-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human bladder urothelial carcinoma tissue; fixative not specified (datasheet A10834-1); 2. Fixation: Image fixative and duration unreported (datasheet A10834-1); 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 A10834-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A10834-1); 8. Primary antibody: Rabbit anti-L3MBTL3, 2-5 μg/ml (datasheet A10834-1); 9. Primary incubation: Overnight at 4 °C (datasheet A10834-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10834-1); Caveat: Staining has low concordance with RNA; verify specificity (HPA tissue IHC); Isoform / epitope: 2 isoforms; check epitope coverage (UniProt); Regulation: Low tissue specificity at RNA level (HPA tissue RNA). Evidence: UniProt Q96JM7, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and nucleoli. In addition…, Datasheet A10834-1, PMC10778146. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/L3MBTL3/immunohistochemistry-ihc-guide.