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LMNB2 / Lamin-B2 · IHC design guide
Design Immunohistochemistry for LMNB2
Plan chromogenic IHC for LMNB2 around the nuclear membranous tissue pattern, with strong staining reported in CNS cells (HPA tissue IHC). Use the catalog antibody’s paraffin-section protocol and compare nuclear rim staining across consistently fixed samples (datasheet M05348; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LMNB2 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 membranous staining in tissue (HPA tissue IHC) | |
| Staining pattern | Nuclear rims in glia and cerebellar granular-layer cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M05348) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
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 M05348) | |
| Caveat | Mitotic lamina disassembly may alter nuclear rim staining (UniProt) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | 0 isoforms listed; residues 618–620 are removed (UniProt) |
Section 1
Recommended LMNB2 IHC & IF Protocols
The catalog antibody protocol uses paraffin sections; the published IHC protocols below report LMNB2 staining in colorectal, breast, and bladder cancer tissues (PMC8007612; PMC7846963; PMC9282643).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet M05348) |
| Fixation | Image fixative and duration unreported (datasheet M05348); 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 M05348); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M05348) |
| Primary antibody | Rabbit monoclonal (clone ADBF-12) anti-LMNB2, 1:50 (datasheet M05348) |
| Primary incubation | Overnight at 4 °C (datasheet M05348) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M05348) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LMNB2-positive staining in glial cells of caudate (HPA tissue IHC: High). HPA tissue profile: Nuclear membranous expression, most abundant in CNS. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet M05348); the three published excerpts do not specify a retrieval method.
Section 2
What Is the Expected LMNB2 Staining Pattern?
LMNB2 is a nuclear lamina protein, so a convincing IHC signal should outline nuclei at their perimeter (UniProt Q03252; HPA: nuclear membranous expression). HPA reports high staining in several CNS cell populations, as well as selected glandular, collecting duct, and endothelial cells (HPA: tissue IHC, Enhanced reliability). LMNB2 has no transmembrane segment; its rim pattern reflects its place in the lamina (UniProt Q03252 topology and function).
| A crisp chromogenic rim surrounds nuclei in glial cells of caudate, cerebral cortex, or hippocampus. | This matches the expected compartment and documented high staining in those cells (UniProt Q03252: nuclear lamina; HPA: High in glial cells at these sites). Score the nuclear rim separately from any diffuse staining. |
| The strongest deposit is cytoplasmic, extracellular, or spread uniformly through nuclei without a discernible rim. | That distribution does not match the reported nuclear membranous pattern (HPA: tissue IHC; UniProt Q03252: nuclear lamina). Check section morphology and detection controls before interpreting it as LMNB2. |
| Strong staining appears mainly in a cell population reported as low, while the expected nuclear rim is absent. | HPA reports low staining in, for example, duodenal glandular cells and colonic endothelial cells (HPA: tissue IHC). Review cell identification and controls; cross-reactivity or endogenous detection activity is possible, but HPA does not establish the cause. |
| Color forms a diffuse haze across tissue, including spaces outside cells. | A tissue-wide haze obscures the nuclear rim used to recognize LMNB2 (HPA: nuclear membranous expression). Assess reagent background and washing with routine IHC controls; haze alone cannot establish target expression. |
| No nuclear rim is visible in a known high-staining compartment, such as kidney collecting ducts. | Collecting ducts are reported as High by HPA (HPA: kidney collecting ducts). Check whether nuclei are preserved and whether a validated positive control stained; one blank section cannot distinguish assay failure from a sample-specific result. |
💡Expected LMNB2 appearanceCall a result positive when a clear nuclear-perimeter stain is present in the assessed cells, especially HPA High populations; diffuse cytoplasmic or extracellular color alone is unconvincing (HPA: tissue IHC; UniProt Q03252: nuclear lamina).
| Cell and tissue context | HPA records High staining in CNS glial cells, cerebellar granular-layer cells, kidney collecting ducts, lung endothelial cells, and epididymal or fallopian-tube glandular cells (HPA: tissue IHC). Its Low entries provide comparison sites, not proven negative controls; HPA lists no negative tissue here. |
| Antibody evidence | The tissue profile has Enhanced reliability, meaning high consistency between staining and RNA data (HPA: tissue IHC). HPA047863 and HPA062477 have Enhanced IHC validation; CAB022593 has Supported IHC validation (HPA: antibody records). These labels do not validate every staining event in a new specimen. |
| Epitope and processing | UniProt lists a lamin-B2 chain spanning residues 1–617 and a propeptide at 618–620, plus modified residues (UniProt Q03252). No antibody epitope is supplied, so these annotations cannot predict which reagent will stain a given section or explain signal loss. |
| Retrieval and detection | No LMNB2-specific antigen-retrieval condition or fixation-sensitivity result is supplied (UniProt Q03252; HPA: supplied records). Treat retrieval and chromogenic detection settings as assay variables to optimize with controls under general IHC practice; do not infer a target-specific fixation effect from HPA staining levels. |
| IF/ICC Q&A: where should signal appear? | A nuclear-membrane rim is the supported IF/ICC location (HPA: subcellular, nuclear membrane supported). HPA lists ICC-IF images in A-431, U-251MG, and U2OS, and Supported ICC validation for HPA062477 (HPA: subcellular and antibody records). These observations do not specify an IF protocol for this IHC guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in the test section or an HPA High positive-control tissue. | The result is indeterminate: the expected population may be missed, nuclear preservation may be poor, or the staining run may have failed (HPA: High tissue populations). | Confirm the relevant cells on the counterstain, then review the positive control and run records. Recheck routine IHC retrieval, reagent, and detection steps without assuming an LMNB2-specific fixation effect. |
| A faint rim appears in a tissue listed as Low. | Low is a reported staining level, not an assertion that LMNB2 is absent (HPA: tissue IHC Low entries). | Compare the same run with an HPA High population and assess whether the faint signal remains confined to nuclear perimeters. Record intensity and cell type rather than calling every faint rim nonspecific. |
| Brown deposit fills cytoplasm or lies outside nuclei. | This conflicts with the reported nuclear membranous location (HPA: tissue IHC; UniProt Q03252: nuclear lamina); the source of the deposit is unresolved. | Inspect morphology and the no-primary control, then review blocking, washing, and chromogen development using routine IHC practice. Require a discernible nuclear rim before assigning LMNB2 positivity. |
| Unexpected cells stain strongly while the anticipated high-staining cells do not. | Cell identification, cross-reactivity, or endogenous detection activity may account for the mismatch; HPA documents specific High and Low populations but cannot diagnose this section (HPA: tissue IHC). | Reidentify the cells on the counterstain and compare a known HPA High tissue on the same run. Use detection controls to investigate background before attributing the unexpected pattern to LMNB2. |
| Diffuse background makes nuclear boundaries hard to score. | Background may arise from routine chromogenic detection or incomplete washing; the haze does not reproduce the reported nuclear rim (HPA: nuclear membranous expression). | Review the no-primary control, blocking and wash steps, and development time. Score only cells whose nuclear edge can be distinguished from surrounding color; document any areas that remain uninterpretable. |
| An IF/ICC image shows a rim but the IHC section lacks one. | HPA supports a nuclear-membrane IF/ICC location, while its tissue IHC profile separately reports nuclear membranous staining (HPA: subcellular and tissue IHC). Agreement in compartment does not establish that both assays will perform alike. | Interpret each preparation against its own controls. For the IHC section, verify the relevant HPA High cells and chromogenic run controls; use the IF/ICC observation only as a location check. |
Experiment evidence
IHC & IF images showing LMNB2 staining
M05348 · IHCIHC analysis of Lamin B2 using anti-Lamin B2 antibody (M05348). Lamin B2 was detected in a paraffin-embedded section of human placenta tissu
M05348 · IHCIHC analysis of Lamin B2 using anti-Lamin B2 antibody (M05348). Lamin B2 was detected in a paraffin-embedded section of human spleen tissue.
M05348 · IFImmunofluorescent analysis of Hela cells, using Lamin B2 Antibody .
PA1636 · IHCIHC analysis of Lamin B2 using anti-Lamin B2 antibody (PA1636). Lamin B2 was detected in a paraffin-embedded section of Rat Intestine tissue
PA1636 · IHCIHC analysis of Lamin B2 using anti-Lamin B2 antibody (PA1636). Lamin B2 was detected in frozen section of rat intestine tissues. The tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000176619-LMNB2/tissue/cerebral+cortex#imid_6040768
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000176619-LMNB2/tissue/colon#imid_17758801
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000176619-LMNB2/tissue/liver#imid_17758802
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000176619-LMNB2/tissue/kidney#imid_17758803
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA062477; green = LMNB2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA062477; green = LMNB2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LMNB2 IHC & IF
🧪Run cerebellum first: cells in the granular layer should show nuclear-rim staining (HPA: High in cells in the granular layer; UniProt Q03252: nuclear lamina). HPA detects LMNB2 in all 45 scored tissues, so there is no validated negative tissue; no cell type on the positive slide is established as negative, and cells without specific staining should lack a defined nuclear rim (HPA: no negative tissue rows; HPA: detected in all 45 scored tissues).
Positive control tissue: Caudate (Glial cells, HPA High)
Negative control tissue: None in HPA: LMNB2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LMNB2 in A-431, U-251MG, U2OS, with annotated localisation: Nuclear membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a concentration-matched rabbit IgG isotype control, then confirm specificity with LMNB2 knockout material or peptide blocking if a suitable peptide is available (selected M05348 caption: rabbit primary; standard IHC controls). Block endogenous peroxidase for DAB detection and assess background in the densely cellular granular layer (selected M05348 caption: peroxidase/DAB detection; HPA: High in cerebellar granular-layer cells).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected M05348 paraffin-section caption does not state its fixative (selected M05348 caption). That caption uses heat retrieval in EDTA at pH 8.0, but it does not establish whether LMNB2 staining depends on retrieval or whether frozen sections or IF are easier (selected M05348 caption). Closely packed granular-layer nuclei may make individual nuclear rims harder to score; use the expected nuclear-lamina pattern when assessing staining (HPA: High in cerebellar granular-layer cells; UniProt Q03252: nuclear lamina; standard IHC interpretation).
HPA tissue IHC evidence for LMNB2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: LMNB2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced LMNB2 IHC Tips
Troubleshoot LMNB2 staining by checking nuclear rim localisation, tissue context and the documented conditions for the IHC-validated antibody.
What retrieval should I try first if LMNB2 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA, pH 8.0 (datasheet M05348). The documented paraffin-section example then uses 10% goat serum, antibody at 1:50 overnight at 4°C, and peroxidase-based DAB detection (datasheet M05348). If the nuclear rim remains faint, check that heating reaches the sections evenly and compare retrieval duration on adjacent sections while holding antibody and detection conditions constant (standard IHC practice). Assess the rim in intact nuclei rather than using diffuse nuclear colour as the endpoint, because LMNB2 belongs to the nuclear lamina (UniProt Q03252; HPA subcellular). Record retrieval duration and tissue integrity alongside staining intensity (standard IHC practice).
Could fixation be responsible for inconsistent LMNB2 staining?
Target-specific LMNB2 sensitivity to fixation is unknown from the supplied evidence; the paraffin-section caption does not state a fixative (datasheet M05348). Compare sections with documented fixation and processing histories before changing retrieval or antibody concentration, since those variables can alter epitope access in IHC (standard IHC practice). On adjacent sections, keep EDTA, pH 8.0 retrieval and the documented 1:50 overnight incubation constant while assessing staining and morphology (datasheet M05348; standard IHC practice). Avoid attributing a faint rim to an LMNB2-specific fixation effect without a controlled comparison, and record the actual fixative and exposure time for each specimen (standard IHC practice).
Where should a convincing LMNB2 IHC signal appear?
Expect a nuclear membranous rim: LMNB2 is a nuclear lamina protein, and HPA reports supported nuclear membrane localisation (UniProt Q03252; HPA subcellular). Evaluate the rim around morphologically intact nuclei at high magnification, distinguishing it from diffuse nucleoplasmic staining or chromogen deposited over the cytoplasm (standard IHC practice). A transmembrane cell-surface pattern would conflict with the record's absence of a transmembrane segment and its nuclear lamina assignment (UniProt Q03252 topology and subcellular location). Use a nuclear counterstain to identify the nuclear boundary, then compare signal across cells on the same section under identical illumination and development conditions (standard IHC practice).
Can epitope position explain differing LMNB2 staining patterns?
The supplied LMNB2 record lists 0 isoforms and identifies an intermediate filament rod at residues 46–402 and an LTD at 462–579 (UniProt Q03252). It also lists modified residues, including phosphorylation at 23, 34, 37 and 316, plus processing of residues 618–620 (UniProt Q03252). The antibody epitope is not specified in the supplied caption, so these features cannot establish why a particular section stains weakly (datasheet M05348; UniProt Q03252). Compare retrieval conditions on adjacent sections and verify the expected nuclear rim before interpreting intensity differences as epitope masking (standard IHC practice; HPA subcellular).
How should I plan a follow-up IF experiment after chromogenic LMNB2 IHC?
Treat IF/ICC as a separate validation workflow; the documented 1:50, overnight, 4°C incubation and EDTA, pH 8.0 retrieval describe paraffin-section IHC (datasheet M05348). For multiplex IF, pair LMNB2 with a marker for the cell population being evaluated, such as glial cells in caudate, where HPA reports high LMNB2 staining (HPA tissue IHC; standard IF practice). Choose a fluorophore channel after checking tissue autofluorescence and include single-colour controls to assess bleed-through (standard IF practice). Because the expected epitope is at the nuclear lamina on the nucleoplasmic side of the inner nuclear membrane, optimise permeabilisation to allow nuclear access without disrupting the rim (UniProt Q03252; standard IF practice).
How can I reduce diffuse DAB background without losing the LMNB2 rim?
First compare the section with a no-primary control to separate antibody-associated signal from staining produced by the detection workflow (standard IHC practice). The documented example uses 10% goat serum, a peroxidase-conjugated secondary for 30 minutes at 37°C, and DAB development (datasheet M05348). Apply a peroxidase block and adjust wash stringency or DAB development using matched sections if background persists; these are general chromogenic IHC steps, not LMNB2-specific findings (standard IHC practice). Judge improvement by preservation of a distinct nuclear rim, the expected LMNB2 location, while noting whether nonspecific colour persists outside nuclei (UniProt Q03252; HPA subcellular; standard IHC practice).
What should I score when comparing LMNB2 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define positivity as a discernible nuclear membranous rim before counting cells, consistent with LMNB2's reported location (UniProt Q03252; HPA subcellular). Report the percentage of rim-positive, morphologically intact nuclei within each predefined cell population; an H-score can additionally capture staining intensity if the same threshold is used throughout (standard IHC practice). Normalise positive-cell counts to all evaluable nuclei in that population, or report rim-positive nuclei per mm² of viable tissue with area measured consistently (standard IHC practice). Keep retrieval, counterstain, DAB development and image settings comparable, and document excluded damaged or necrotic regions before comparing specimens (standard IHC practice).
How do I distinguish true LMNB2 staining from section artefacts?
Give greatest weight to a nuclear rim in intact cells, matching LMNB2's nuclear lamina assignment and HPA's nuclear membranous profile (UniProt Q03252; HPA tissue IHC). Interpret cell populations separately: HPA reports high staining in caudate glial cells but low staining in placental decidual cells, while the catalog example demonstrates detection in a paraffin section of placenta (HPA tissue IHC; datasheet M05348). Treat isolated colour at section edges, necrotic areas or outside nuclear boundaries cautiously and compare it with a no-primary control (standard IHC practice). Check whether residual endogenous peroxidase accounts for DAB colour before calling an unexpected cellular pattern LMNB2-positive (standard IHC practice).
Boster reagents
Best LMNB2 / Lamin-B2 IHC Antibodies
LMNB2 antibodies have paraffin-section IHC images from human placenta and rat intestine, and an IF image from HeLa cells (M05348 and PA1636 IHC captions; M05348 IF caption).
Real IHC data
Anti-Lamin B2 Monoclonal Antibody
Cat # M05348
Real IHC data
Anti-Lamin B2/LMNB2 Antibody ®
Cat # PA1636
M05348 has IHC images from human placenta, spleen, breast cancer and lung cancer paraffin sections, plus an IF image from HeLa cells (M05348 image captions). PA1636 has IHC images from rat intestine paraffin and frozen sections and a human brain paraffin section (PA1636 image captions).
Which to pick: For tissue IHC across human and rat samples, choose PA1636: its images cover human brain and rat intestine paraffin sections, and it lists Human, Mouse and Rat reactivity (PA1636 IHC captions; PA1636 catalog reactivity). For IF/ICC, choose the rabbit monoclonal M05348: IF and ICC are listed applications, and its IF caption shows HeLa cells (M05348 catalog applications and clone; M05348 IF caption). M05348 also has human paraffin-section IHC images and lists Human, Mouse and Rat reactivity; neither SKU’s paraffin-section caption reports a fixative (M05348 catalog reactivity and IHC captions; PA1636 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q03252 (LMNB2_HUMAN, Lamin-B2).
- Human Protein Atlas. LMNB2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LMNB2 subcellular location (ICC-IF): Localized to the nuclear membrane..
- Human Protein Atlas. LMNB2 antibody validation summary (3 antibodies).
- LMNB2 promotes the progression of colorectal cancer by silencing p21 expression. Cell death & disease 2021 — PMC8007612.
- Lamin B2 promotes the progression of triple negative breast cancer via mediating cell proliferation and apoptosis. Bioscience reports 2021 — PMC7846963.
- Lamin B2 contributes to the proliferation of bladder cancer cells via activating the expression of cell division cycle‑associated protein 3. International journal of molecular medicine 2022 — PMC9282643.
- Identification of disulfidptosis-related molecular subtypes in pleural mesothelioma demonstrates a correlation with the immune infiltration, prognosis and therapy efficacy. Discover oncology 2026 — PMC13498538.
- PubMed PMID:15057824 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:1630457 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LMNB2 (Lamin-B2). Expected localisation Nuclear membranous staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M05348). Validated IHC antibody: M05348 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Glial cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human placenta tissue; fixative not specified (datasheet M05348); 2. Fixation: Image fixative and duration unreported (datasheet M05348); 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 M05348); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M05348); 8. Primary antibody: Rabbit monoclonal (clone ADBF-12) anti-LMNB2, 1:50 (datasheet M05348); 9. Primary incubation: Overnight at 4 °C (datasheet M05348). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M05348); Caveat: Mitotic lamina disassembly may alter nuclear rim staining (UniProt); Isoform / epitope: 0 isoforms listed; residues 618–620 are removed (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt Q03252, HPA tissue IHC · Enhanced, HPA ICC-IF · nuclear membrane., Datasheet M05348, PMC8007612, PMC7846963. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LMNB2/immunohistochemistry-ihc-guide.