This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
LSM8 / U6 snRNA-associated Sm-like protein LSm8 · IHC design guide
Design Immunohistochemistry for LSM8
This guide helps plan chromogenic IHC for LSM8, which shows general nuclear staining in tissue (HPA tissue IHC). Adrenal gland glandular cells show high staining, while ovarian stromal cells have no detected staining (HPA tissue IHC); interpret this comparison in light of the uncertain staining reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LSM8 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 | General nuclear staining observed (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across diverse tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A10947) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Caudate+2 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 | Staining and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | No specific regulation annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; epitope map unknown (UniProt) |
Section 1
Recommended LSM8 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet A10947). The published gastric tissue IHC workflow below selected samples by LSM8 expression but stained immune cell markers (PMC10064066).
| Sample | Paraffin-embedded human skin cancer tissue; fixative not specified (datasheet A10947) |
| Fixation | Image fixative and duration unreported (datasheet A10947); 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 A10947); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A10947) |
| Primary antibody | Rabbit anti-LSM8, 1-2μg/ml (datasheet A10947) |
| Primary incubation | Overnight at 4 °C (datasheet A10947) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A10947) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LSM8-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 (datasheet A10947); the published gastric tissue workflow also used EDTA at pH 8.0 (PMC10064066).
Section 2
What Is the Expected LSM8 Staining Pattern?
LSM8 is a nuclear spliceosome component without a transmembrane segment (UniProt O95777). In paraffin sections, expect mainly nuclear staining across several cell types, with high staining reported in glandular cells of the adrenal, parathyroid and thyroid glands and in bronchial respiratory epithelium (HPA: tissue IHC). Treat these as reference patterns, since the tissue IHC reliability is Uncertain, with medium consistency between antibody staining and RNA expression (HPA: reliability).
| Clear nuclear staining in glandular cells of the thyroid or parathyroid gland. | This fits the reported high staining in those cells (HPA: tissue IHC) and the nuclear location of LSM8 (UniProt O95777). Compare cells within the same section before judging intensity. |
| Predominantly cytoplasmic or membranous chromogen, with little nuclear signal. | This conflicts with the general nuclear IHC profile (HPA: tissue IHC) and nuclear annotation (UniProt O95777). Review morphology and controls; nonspecific staining or detection artefact may explain the pattern (general IHC practice). |
| Strong staining in ovarian stromal cells or soft tissue fibroblasts. | Those cells were reported as not detected (HPA: tissue IHC). If staining persists after controls, consider antibody cross-reactivity or endogenous detection activity (general IHC practice); HPA's Uncertain reliability limits a definitive negative call. |
| Haze or chromogen deposits across nuclei and surrounding tissue. | A diffuse pattern is difficult to score against the expected nuclear distribution (HPA: tissue IHC). Check the no-primary control, blocking and detection reagents for background (general IHC practice). |
| No nuclear signal in thyroid glandular cells. | HPA reports high staining in these cells, so the result warrants a technical check (HPA: tissue IHC). Confirm tissue integrity, reagent performance and scoring against a concurrent control (general IHC practice); the HPA result is not a guaranteed positive. |
💡Expected LSM8 appearanceCall positive when glandular or bronchial epithelial cells show distinct, mainly nuclear staining, potentially high in the reported examples (HPA: tissue IHC); diffuse tissue staining or dominant membrane signal is suspect against the nuclear reference pattern (UniProt O95777; general IHC practice).
| Tissue and cell selection | High staining was reported in adrenal, cervical, gallbladder, parathyroid and thyroid glandular cells, bronchial respiratory epithelium and testicular Leydig cells (HPA: tissue IHC). Score the named cells, rather than an entire organ, against that reference. |
| Lower-signal comparisons | Cerebellar granular-layer cells were medium; several listed neuronal, glial and glandular populations were low (HPA: tissue IHC). A weak result in those populations alone gives less information about assay performance. |
| Antibody evidence | Tissue IHC reliability is Uncertain, and the listed antibody HPA020116 is rated Uncertain for IHC (HPA: reliability; HPA: antibody validation). Interpret agreement with the atlas pattern as supportive, not proof of specificity. |
| Molecular location | LSM8 is nuclear, lacks a transmembrane segment and participates in the LSM2–8 spliceosomal complex (UniProt O95777). These annotations support a nuclear reading; they do not establish antigen retrieval conditions or fixation sensitivity. |
| IF/ICC Q: What location is reported? | A: Nucleoplasm and vesicles are approved locations in ICC-IF images, including A-431 cells (HPA: subcellular; HPA: antibody validation). That IF observation does not make vesicular chromogen a required tissue IHC result. |
| Situation | Likely cause | Next action |
|---|---|---|
| Nuclei are blank in a reported high-staining tissue. | Assay failure or an unrepresentative section is possible (general IHC practice); HPA tissue IHC is Uncertain (HPA: reliability). | Check tissue morphology and a concurrent control, then review the existing retrieval and detection workflow (general IHC practice). No LSM8-specific fixation sensitivity is reported here. |
| Cytoplasmic staining dominates the slide. | The distribution conflicts with nuclear LSM8 (UniProt O95777) and general nuclear tissue IHC (HPA: tissue IHC). Background or misidentified cell boundaries may contribute (general IHC practice). | Inspect counterstained nuclei and compare a no-primary control; reassess any signal that remains outside nuclei (general IHC practice). |
| Negative-comparison cells stain strongly. | Ovarian stromal cells and soft tissue fibroblasts were not detected in the atlas (HPA: tissue IHC). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Compare the same cell types with no-primary and detection controls, and confirm cell identity before calling the signal specific (general IHC practice). |
| Every region has diffuse brown background. | A broad deposit obscures the expected nuclear distribution (HPA: tissue IHC); inadequate blocking, washing or detection control may contribute (general IHC practice). | Inspect the no-primary control and review blocking, washes and chromogen development within the established assay (general IHC practice). |
| Only weak signal appears in a low-staining population. | Some neuronal, glial and glandular populations were reported low (HPA: tissue IHC), so their weak signal alone cannot establish assay failure. | Assess a concurrently processed tissue and cell population reported high by HPA, while keeping the Uncertain IHC reliability in mind (HPA: tissue IHC; HPA: reliability). |
| Vesicular signal appears in tissue IHC. | Vesicles are approved in ICC-IF (HPA: subcellular), while the tissue IHC profile is generally nuclear (HPA: tissue IHC). The methods provide different evidence. | Score the nuclear tissue pattern first; use tissue controls to assess vesicular chromogen before assigning it to LSM8 (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing LSM8 staining
A10947 · IHCIHC analysis of LSM8 using anti-LSM8 antibody (A10947). LSM8 was detected in paraffin-embedded section of human skin cancer tissue. Heat med
A10947 · IHCIHC analysis of LSM8 using anti-LSM8 antibody (A10947). LSM8 was detected in paraffin-embedded section of mouse testis tissue. Heat mediated
A10947 · IFIF analysis of LSM8 using anti-LSM8 antibody (A10947). LSM8 was detected in immunocytochemical section of A431 cells. Enzyme antigen retriev
M10947 · IHCIHC analysis of LSM8 using anti-LSM8 antibody (M10947). LSM8 was detected in a paraffin-embedded section of human renal clear cell carcinoma
M10947 · IHCIHC analysis of LSM8 using anti-LSM8 antibody (M10947). LSM8 was detected in a paraffin-embedded section of human colonic adenocarcinoma tis
M10947 · IFIF analysis of LSM8 using anti-LSM8 antibody (M10947). LSM8 was detected in an immunocytochemical section of Hela cells. Enzyme antigen retr
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000128534-LSM8/tissue/cerebral+cortex#imid_5526160
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000128534-LSM8/tissue/colon#imid_5526078
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000128534-LSM8/tissue/liver#imid_5526071
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000128534-LSM8/tissue/kidney#imid_5526080
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA020116; green = LSM8, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA020116; green = LSM8, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LSM8 IHC & IF
🧪Run adrenal gland first: its glandular cells should stain (HPA: High in adrenal gland glandular cells). Use caudate glial cells as the negative tissue reference (HPA: Not detected in caudate glial cells); on the adrenal slide, cells without nuclear staining can help gauge background but are not validated LSM8-negative cells (UniProt O95777: nucleus).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LSM8 in A-431, with annotated localisation: Nucleoplasm (approved), Vesicles (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), a rabbit immunoglobulin isotype control matched to the primary antibody’s clonality, and LSM8-knockout material or a peptide-block control if available (caption: rabbit anti-LSM8 antibody). For the adrenal slide, block endogenous peroxidase and biotin and assess residual background because the documented IHC detection uses a biotin-based complex with DAB (caption: SABC/DAB detection).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A10947 paraffin-section caption does not state a fixative (caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 is documented for that IHC example, but whether retrieval is required has not been established (caption: heat-mediated EDTA retrieval). The supplied evidence does not establish that frozen sections or IF are easier; assess endogenous peroxidase and biotin background in the adrenal section when using the documented SABC/DAB method (caption: SABC/DAB detection).
HPA tissue IHC evidence for LSM8
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced LSM8 IHC Tips
Troubleshoot LSM8 staining by checking nuclear localisation, section quality and controls before interpreting differences in chromogenic signal (UniProt O95777; HPA tissue IHC).
What retrieval should I use when LSM8 staining is weak in paraffin sections?
Start with heat-mediated EDTA at pH 8.0 for paraffin sections (datasheet A10947). The selected tissue image used this retrieval before overnight incubation at 4°C with 2 µg/mL catalog antibody (datasheet A10947). If nuclear staining remains weak, compare retrieval heating times on adjacent sections while keeping antibody concentration, development time and counterstain constant (standard IHC practice). Include a no-primary control to distinguish increased antigen signal from retrieval-related background (standard IHC practice). Score staining in viable nuclei, since nuclear localisation is expected and the tissue IHC profile describes general nuclear expression (UniProt O95777; HPA tissue IHC).
How should I assess whether fixation is affecting LSM8 staining?
Target-specific fixation sensitivity is unknown; the selected image identifies paraffin-embedded tissue but does not report its fixative (datasheet A10947). Record fixative, fixation duration and processing history for each section before comparing staining intensity (standard IHC practice). If specimens have different fixation histories, stain matched sections in one run with the same EDTA pH 8.0 retrieval and 2 µg/mL antibody starting condition (datasheet A10947; standard IHC practice). Compare viable nuclear signal with no-primary controls and review tissue preservation before changing retrieval or antibody concentration (UniProt O95777; standard IHC practice). Do not attribute a weak result to an LSM8-specific fixation effect without a controlled comparison (standard IHC practice).
Should vesicular or cytoplasmic staining count as LSM8 positive?
Prioritise staining within morphologically intact nuclei: UniProt places LSM8 in the nucleus, and tissue IHC reports general nuclear expression (UniProt O95777; HPA tissue IHC). HPA cell imaging also lists nucleoplasm and vesicles as approved locations, so a vesicular pattern warrants review rather than automatic rejection (HPA subcellular). Segment nuclei using the counterstain and score nuclear DAB separately from cytoplasmic or punctate signal (standard IHC practice). Check whether puncta recur in the same cell compartment across adjacent sections and disappear in no-primary controls (standard IHC practice). Avoid assigning diffuse cytoplasmic DAB to LSM8 from colour alone when the expected tissue pattern is nuclear (HPA tissue IHC; standard IHC practice).
Could LSM8 processing or its epitope explain inconsistent nuclear staining?
The record lists 0 isoforms, an Sm domain at residues 1–76, and a processed chain spanning residues 2–96 (UniProt O95777). It also lists N-terminal acetylation at residue 2, no glycosylation sites and no transmembrane segment (UniProt O95777). The catalog antibody’s precise epitope is not supplied, so those features cannot establish whether fixation or retrieval masks antibody binding (datasheet A10947; UniProt O95777). Compare nuclear staining across adjacent sections using the documented EDTA pH 8.0 retrieval before changing detection conditions (datasheet A10947; standard IHC practice). If an independent antibody is available, compare its nuclear pattern while documenting its own validation and epitope information (standard IHC practice).
How can I cross-check LSM8 IHC with multiplex immunofluorescence?
Use IF as a separate localisation check: HPA cell imaging includes A-431 images and lists nucleoplasm and vesicles as approved LSM8 locations (HPA subcellular). For respiratory epithelium, pair LSM8 with an epithelial marker such as cytokeratin and a nuclear counterstain to identify the expected cell population (HPA tissue IHC: high in respiratory epithelial cells; standard IF practice). Choose a far-red detection channel when tissue autofluorescence obscures shorter-wavelength signal, and inspect single-stain controls before interpreting overlap (standard IF practice). Because the expected main signal is nuclear and LSM8 has no transmembrane segment, optimise permeabilisation for access across the nuclear envelope; 0.1% Triton X-100 is a general starting condition, not an LSM8-validated setting (UniProt O95777; standard IF practice).
How do I distinguish LSM8 DAB signal from chromogenic background?
The selected image used 10% goat serum blocking, biotinylated secondary antibody, a streptavidin–biotin detection complex and DAB (datasheet A10947). Run a no-primary section through the same detection steps to reveal background from secondary reagents, endogenous biotin or chromogen deposition (standard IHC practice). Include a peroxidase block before DAB development and check whether staining persists in blood-rich or damaged areas on the control section (standard IHC practice). If background rises with retrieval, compare adjacent sections while holding the 2 µg/mL primary concentration and DAB development time constant (datasheet A10947; standard IHC practice). Assess nuclear signal against local background within viable tissue, rather than judging the darkest area alone (UniProt O95777; standard IHC practice).
What should I measure when comparing LSM8 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score viable nuclei because the tissue IHC profile describes general nuclear expression and UniProt assigns LSM8 to the nucleus (HPA tissue IHC; UniProt O95777). Record the percentage of positive nuclei and intensity on a 0–3 scale; their weighted H-score spans 0–300 (standard IHC scoring practice). Normalise positive counts to all evaluable nuclei in the same cell population, or report positive-cell density per mm² of viable tissue (standard IHC scoring practice). Keep retrieval, antibody concentration, DAB development and image thresholds consistent across comparison groups (standard IHC practice). Report cell type and section quality alongside the score because HPA tissue staining has uncertain reliability and only medium agreement with RNA expression (HPA tissue IHC).
When is an apparent LSM8-positive section likely to be misleading?
A convincing result shows reproducible nuclear staining in intact cells, consistent with UniProt localisation and the general nuclear tissue IHC profile (UniProt O95777; HPA tissue IHC). Compare the stained cell type with HPA observations: respiratory epithelial cells are high, whereas ovarian stroma cells are listed as not detected (HPA tissue IHC). Treat strong staining confined to cut edges, necrotic areas or poorly preserved cells as suspect and compare it with interior viable tissue (standard IHC practice). Check no-primary and peroxidase-block controls before assigning DAB deposits to LSM8 (standard IHC practice). Interpret unexpected compartment or cell-type patterns cautiously because the HPA tissue IHC reliability is uncertain (HPA tissue IHC).
Boster reagents
Best LSM8 / U6 snRNA-associated Sm-like protein LSm8 IHC Antibodies
Anti-LSM8 antibodies have paraffin-section IHC images from human tissues and mouse and rat testis, plus ICC/IF images from A431 and HeLa cells (catalog image captions).
Real IHC data
Anti-LSM8 Antibody ®
Cat # A10947
Real IHC data
Anti-LSM8 Antibody ® (monoclonal, 6B11)
Cat # M10947
A10947 has IHC images from human skin and ovary cancers and mouse and rat testis, plus an ICC/IF image from A431 cells (A10947 image captions). M10947 has IHC images from human renal and colonic cancers, placenta, and spleen, plus an ICC/IF image from HeLa cells (M10947 image captions).
Which to pick: For paraffin-section tissue IHC across human, mouse, and rat samples, choose rabbit antibody A10947: its IHC captions show all three species, and its listed reactivity includes all three (A10947 IHC captions; catalog reactivity). For human tissue IHC where a monoclonal is preferred, choose mouse clone 6B11, M10947; its IHC captions show human paraffin sections, while A10947 has the stronger cross-species IHC image record (M10947 catalog clone and IHC captions; A10947 IHC captions). Both list IF/ICC and have cell images, so choose A10947 for A431 or M10947 for HeLa as the documented starting point; the fixative is unreported for both paraffin-section IHC examples (catalog applications; A10947 and M10947 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95777 (LSM8_HUMAN, U6 snRNA-associated Sm-like protein LSm8).
- Human Protein Atlas. LSM8 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. LSM8 subcellular location (ICC-IF): Localized to the nucleoplasm and vesicles..
- Human Protein Atlas. LSM8 antibody validation summary (1 antibodies).
- Identification of LSM family members as potential chemoresistance predictive and therapeutic biomarkers for gastric cancer. Frontiers in oncology 2023 — PMC10064066.
- PubMed PMID:10523320 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LSM8 (U6 snRNA-associated Sm-like protein LSm8). Expected localisation General nuclear staining observed (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A10947). Validated IHC antibody: A10947 at 1-2μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Caudate. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human skin cancer tissue; fixative not specified (datasheet A10947); 2. Fixation: Image fixative and duration unreported (datasheet A10947); 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 A10947); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A10947); 8. Primary antibody: Rabbit anti-LSM8, 1-2μg/ml (datasheet A10947); 9. Primary incubation: Overnight at 4 °C (datasheet A10947). Watch for: Fixation: Keep fixation conditions consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; epitope map unknown (UniProt); Regulation: No specific regulation annotated (UniProt). Evidence: UniProt O95777, HPA tissue IHC · Uncertain, HPA ICC-IF · nucleoplasm and vesicles., Datasheet A10947, PMC10064066. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LSM8/immunohistochemistry-ihc-guide.