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LAMB2 / Laminin subunit beta-2 · IHC design guide
Design Immunohistochemistry for LAMB2
Plan LAMB2 paraffin IHC around its extracellular matrix and basement membrane localisation (UniProt) and the cytoplasmic staining reported in selected tissues (HPA tissue IHC). Use high staining in breast adipocytes or muscle cells as a control reference (HPA tissue IHC), with the catalog antibody's 1:50–1:200 IHC dilution (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LAMB2 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 | ECM and basement membrane; cytoplasmic tissue staining (UniProt; HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in select cells; prominent in ECM and vascular smooth muscle (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Adrenal gland+4 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 | Secreted ECM deposition may differ from cellular staining (UniProt) | |
| Regulation | Expression regulators unreported (UniProt) | |
| Isoform / epitope | No isoforms annotated; mature chain 33–1798 has no cytoplasmic epitope (UniProt) |
Section 1
Recommended LAMB2 IHC & IF Protocols
The catalog antibody protocol is accompanied by published LAMB2 IHC methods for rabbit aortic arches (PMC13409397) and patient and control kidney sections (PMC7048679).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A08306-1) |
| Fixation | Image fixative and duration unreported (datasheet A08306-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-LAMB2, 1:50-1:200 (datasheet A08306-1) |
| 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 | LAMB2-positive staining in adipocytes of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in selected tissues, most abundant in extracellular matrix and vascular smooth muscles. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval at 95–98 °C for 20 min (page retrieval); the rabbit aorta protocol also reports thermal retrieval (PMC13409397).
Section 2
What Is the Expected LAMB2 Staining Pattern?
LAMB2 is a secreted basement-membrane and extracellular-matrix protein with no transmembrane segment (UniProt P55268). In paraffin-section IHC, expect extracellular matrix-associated staining, with HPA also reporting cytoplasmic expression in selected tissues and abundant staining in vascular smooth muscle (HPA tissue IHC: Supported). High staining is reported in breast adipocytes, cardiomyocytes, skeletal myocytes, and smooth muscle cells (HPA tissue IHC).
| Matrix-associated staining and staining in the expected cells, with stronger signal in muscle or breast adipocytes. | This fits the secreted basement-membrane location (UniProt P55268) and HPA's high staining in cardiomyocytes, skeletal myocytes, smooth muscle cells, and breast adipocytes (HPA tissue IHC). Assess the structure and cell being stained, rather than intensity alone (general IHC practice). |
| Strong nuclear staining dominates the slide. | A dominant nuclear pattern does not fit the secreted extracellular location (UniProt P55268) or HPA's tissue description of cytoplasmic and matrix-associated staining (HPA tissue IHC). Treat it as a possible artefact and check controls before assigning it to LAMB2 (general IHC practice). |
| Strong cellular staining appears in cells reported as undetected, such as bone-marrow hematopoietic cells. | That cell-level result conflicts with the HPA observation for hematopoietic cells (HPA tissue IHC: Not detected). Cross-reactivity or endogenous chromogenic detection activity is possible; check the antibody and detection controls (general IHC practice). The HPA cell call does not rule out nearby extracellular staining. |
| Diffuse chromogen covers tissue and empty areas, obscuring structures. | Uniform haze cannot reliably establish the matrix-associated pattern reported for LAMB2 (HPA tissue IHC; UniProt P55268). Background can arise from detection chemistry or nonspecific binding; compare a no-primary control and review the general blocking, washing, and detection workflow (general IHC practice). |
| No signal appears in a known-positive control section. | Check whether the sampled structure is one HPA reports as positive: breast adipocytes, cardiomyocytes, skeletal myocytes, and smooth muscle cells are high, while glomerular cells are low (HPA tissue IHC). A failed positive control makes the slide uninterpretable; verify section quality and the general IHC detection workflow. |
💡Expected LAMB2 appearanceCall positive when chromogen follows basement-membrane or extracellular-matrix structures (UniProt P55268; HPA tissue IHC) and expected cells show the reported level, such as high staining in smooth muscle cells or cardiomyocytes (HPA tissue IHC); dominant nuclear staining is discordant with these locations.
| Extracellular location | LAMB2 is secreted into basement membrane and extracellular matrix, without a transmembrane segment (UniProt P55268). Interpret staining along tissue structures as well as staining assigned to cells; do not require a cell-surface outline. |
| Tissue and cell choice | HPA reports high staining in cardiomyocytes, skeletal myocytes, smooth muscle cells, and breast adipocytes, but low staining in glomerular cells (HPA tissue IHC). Select and score controls by the named cell or structure; a weak glomerular-cell result alone is a limited positive control. |
| Protein processing | UniProt annotates a signal peptide at residues 1–32 and a mature chain at 33–1798, with no propeptide (UniProt P55268). The payload gives no antibody epitope, so it cannot predict whether this processing changes the catalog antibody's IHC signal. |
| Antibody validation | HPA lists IHC as Supported for HPA001895, CAB000053, and CAB078160 (HPA antibodies). That status supports their reported IHC use; it does not validate an unspecified catalog antibody or establish that every tissue and compartment will stain identically. |
| Antigen retrieval | The payload gives no LAMB2-specific retrieval condition or fixation-sensitivity result. If the positive control fails, review the antibody's validated IHC-P procedure and include a known-positive section when assessing a general retrieval adjustment (general IHC practice). |
| IF/ICC Q: Should its pattern be scored like paraffin IHC? | A: HPA reports approved vesicular localization, with additional cytosol staining, in ICC-IF (HPA subcellular). UniProt places secreted LAMB2 in extracellular matrix (UniProt P55268). Interpret each assay in its own context; the ICC-IF observation is not an IHC-P scoring rule. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive muscle section is blank. | This conflicts with HPA's high staining in cardiomyocytes, skeletal myocytes, and smooth muscle cells (HPA tissue IHC); the cause cannot be assigned from the tissue result alone. | Confirm the relevant cells are present, then check primary-antibody application, chromogenic detection, and the general IHC-P workflow with the positive control (general IHC practice). |
| Glomerular cells stain weakly. | Low glomerular-cell staining is reported by HPA (HPA tissue IHC: Kidney, cells in glomeruli, Low). | Score the glomerular cells separately from surrounding structures; compare with a high-staining tissue before calling the assay failed (HPA tissue IHC; general IHC practice). |
| An expected negative cell population stains strongly. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA reports no staining in bone-marrow hematopoietic cells and several specified glandular populations (HPA tissue IHC). | Identify the stained cell type precisely and review a no-primary control. Recheck specificity before treating cellular staining as LAMB2 (general IHC practice). |
| The slide has widespread brown haze. | Background from nonspecific binding or chromogenic detection can obscure an interpretable matrix pattern (general IHC practice; UniProt P55268 location). | Compare the no-primary control, then review general blocking, washing, and detection steps; reassess only where tissue structures remain distinguishable (general IHC practice). |
| Nuclear signal is stronger than matrix or expected-cell signal. | Nuclear dominance is discordant with UniProt's secreted extracellular location and HPA's matrix, vascular smooth-muscle, and selected cytoplasmic observations (UniProt P55268; HPA tissue IHC). | Check whether the same nuclear signal appears in controls and confirm the antibody's IHC validation before interpreting it as target staining (general IHC practice). |
| ICC-IF shows vesicles while IHC-P emphasizes tissue matrix. | HPA reports vesicles and additional cytosol in ICC-IF, whereas its tissue IHC profile emphasizes extracellular matrix and vascular smooth muscle (HPA subcellular; HPA tissue IHC). | Record the assay and compartment with each observation. Use the tissue IHC evidence to interpret the paraffin section and the ICC-IF evidence to describe the cellular image. |
Experiment evidence
IHC & IF images showing LAMB2 staining
A08306-1 · IHCImmunohistochemistry (IHC) analyzes of Laminin β-2 (C91) pAb in paraffin-embedded human breast carcinoma tissue at 1:100.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000172037-LAMB2/tissue/cerebral+cortex#imid_137010
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000172037-LAMB2/tissue/colon#imid_136944
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000172037-LAMB2/tissue/liver#imid_136945
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000172037-LAMB2/tissue/kidney#imid_136954
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA050033; green = LAMB2, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA050033; green = LAMB2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LAMB2 IHC & IF
🧪Run breast first and assess staining around adipocytes (HPA: High in breast adipocytes; UniProt P55268: basement-membrane localisation). Use bone marrow hematopoietic cells as the negative comparison (HPA: Not detected); on the breast slide, treat non-adipocyte areas lacking specific basement-membrane staining as internal background, without assuming every other breast cell is negative (UniProt P55268: basement-membrane localisation).
Positive control tissue: Breast (Adipocytes, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LAMB2 in HeLa, U-251MG, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species and clonality; and LAMB2-knockout tissue as a biological negative where available (standard IHC practice). Quench endogenous peroxidase for chromogenic detection and assess breast tissue background on the no-primary slide (standard IHC practice).
⚠️Feasibility: The selected A08306-1 caption reports paraffin-section breast carcinoma IHC at 1:100, but its fixative is unreported (selected-SKU tissue-IHC caption). A target-specific fixation window, fixation effect, and retrieval dependency are unreported in the supplied evidence; retrieval therefore needs empirical optimization (supplied target/application evidence). Frozen sections and IF are not established as easier by these sources; interpret staining against the expected extracellular basement-membrane pattern and check adipose-rich areas for background (UniProt P55268 localisation; standard IHC practice).
HPA tissue IHC evidence for LAMB2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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 |
|---|---|---|---|---|
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | High | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LAMB2 IHC Tips
Use the IHC evidence and LAMB2 localisation to assess staining in paraffin sections; treat IF findings as a separate application.
What retrieval should I start with for LAMB2 in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 minutes (page retrieval specification). Allow sections to cool in buffer, then compare staining with a control section processed in the same run (standard IHC practice). LAMB2 is a secreted basement-membrane protein, so assess whether retrieval reveals coherent extracellular outlines rather than relying on total brown area alone (UniProt P55268 localisation). If staining remains weak, test an alternative retrieval condition on adjacent sections while keeping antibody and detection settings constant (standard IHC practice). Record tissue preservation alongside signal, since harsh retrieval can make extracellular patterns difficult to judge (standard IHC practice).
How should I troubleshoot weak LAMB2 staining when fixation details are missing?
The selected image documents paraffin-embedded human breast carcinoma at 1:100, but its caption does not state the fixative (A08306-1 caption). Target-specific fixation sensitivity is therefore unknown; neither tissue staining patterns nor protein features establish which fixation condition preserves this antibody’s epitope (A08306-1 caption; HPA tissue IHC; UniProt P55268). Compare sections with documented processing histories and run them through the same retrieval, antibody incubation and chromogenic detection steps (standard IHC practice). Include a positive control with recognizable extracellular staining and inspect morphology before calling a faint result negative (UniProt P55268 localisation; standard IHC practice). Record fixation details when available so later runs can be compared fairly (standard IHC practice).
Where should convincing LAMB2 staining appear in tissue IHC?
Expect staining associated with extracellular matrix and basement membranes because LAMB2 is secreted and has no transmembrane segment (UniProt P55268 localisation and topology). The tissue profile reports strongest abundance in extracellular matrix and vascular smooth muscles, while selected cells show cytoplasmic staining (HPA tissue IHC). Evaluate outlines around structures and vessels separately from cytoplasmic signal, using the section’s anatomy to decide whether the distribution is plausible (UniProt P55268 localisation; HPA tissue IHC; standard IHC practice). LAMB2 is also concentrated in the neuromuscular junction synaptic cleft, a specialised extracellular location (UniProt P55268 localisation). Diffuse nuclear staining alone warrants investigation with controls before biological interpretation (UniProt P55268 localisation; standard IHC practice).
Could epitope accessibility explain inconsistent LAMB2 staining?
The record lists 0 annotated isoforms, but that does not identify the catalog antibody’s epitope or establish that every tissue preparation exposes it (UniProt P55268 isoforms; supplied product caption). LAMB2 is processed from a precursor with a 1–32 signal peptide to a 33–1798 chain, so an epitope’s position matters when interpreting staining (UniProt P55268 processing). It also contains laminin domains, disulfide-linked complex structure and seven annotated glycosylation sites that may affect accessibility (UniProt P55268 domains, subunit and glycosylation). Check the antibody’s documented immunogen region before attributing weak staining to a particular modification (standard IHC practice). Compare controlled retrieval conditions on adjacent sections if accessibility remains a concern (standard IHC practice).
How can IF help resolve ambiguous LAMB2 localisation?
For a separate IF experiment, multiplex LAMB2 with a smooth-muscle cell marker to relate signal to the vascular smooth-muscle pattern reported in tissue (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and place the weaker channel away from the tissue’s strongest autofluorescence; inspect unstained and single-stained controls first (standard IF practice). For an extracellular epitope, begin without detergent permeabilisation, since mature LAMB2 is secreted and lacks a transmembrane segment (UniProt P55268 localisation and topology; standard IF practice). If testing a cytosolic or vesicular signal, evaluate gentle permeabilisation separately because HPA reports those IF locations (HPA subcellular; standard IF practice). Do not transfer the paraffin IHC caption into an IF validation claim (A08306-1 caption).
What should I check when LAMB2 chromogenic staining looks diffuse?
First compare the stained section with a no-primary control to identify signal from the detection system, and check whether brown precipitate follows tissue edges or damaged areas (standard IHC practice). Include a peroxidase block before chromogenic detection and assess residual endogenous enzyme staining with the control (standard IHC practice). LAMB2 can occupy extracellular matrix and basement membranes, so broad extracellular staining can be genuine when it follows coherent anatomy (UniProt P55268 localisation). If background persists, titrate the primary antibody around the image-caption condition of 1:100 while holding retrieval and detection constant (A08306-1 caption; standard IHC practice). Compare the same anatomical regions across runs before adjusting interpretation (standard IHC practice).
How should I score LAMB2 staining when much of it is extracellular? ⚠ ANSWER MARKED FOR VERIFICATION
Define anatomical regions before scoring and measure basement-membrane or extracellular staining as positive area or signal density per mm² of eligible tissue (UniProt P55268 localisation; standard image-analysis practice). Normalise to the relevant tissue area or number of assessable structures, excluding folds, necrosis and blank space by a prespecified rule (standard IHC quantification practice). If reporting cytoplasmic staining in a defined cell population, record percent positive cells or an H-score separately from the extracellular measurement (HPA tissue IHC; standard IHC scoring practice). Keep retrieval, exposure, chromogen development and threshold settings consistent across sections (standard IHC quantification practice). Report the compartments separately so a change in tissue composition does not masquerade as altered LAMB2 expression (UniProt P55268 localisation; standard IHC quantification practice).
How do I distinguish LAMB2 signal from artefact in a paraffin section?
Give greatest weight to staining that follows basement membranes or organised extracellular matrix, consistent with secreted LAMB2 (UniProt P55268 localisation). HPA also reports vascular smooth-muscle abundance and cytoplasmic staining in selected tissues, so judge those patterns against local anatomy rather than requiring one appearance everywhere (HPA tissue IHC). Treat isolated nuclear colour, section-edge accentuation and necrotic debris as suspect until controls and adjacent intact tissue support the pattern (UniProt P55268 localisation; standard IHC practice). A no-primary section helps reveal endogenous enzyme or detection background, particularly when interpreting diffuse brown staining (standard IHC practice). Finally, distinguish a low cell-associated score from absent extracellular deposition when reporting the result (UniProt P55268 localisation; standard IHC scoring practice).
Boster reagents
Best LAMB2 / Laminin subunit beta-2 IHC Antibodies
A08306-1 has an IHC image from paraffin-embedded human breast carcinoma tissue (catalog image caption); listed reactivity is human, mouse and rat (catalog). No IF image is supplied (catalog).
Real IHC data
Anti-Laminin Beta-2 (C91) LAMB2 Antibody
Cat # A08306-1
A08306-1 is listed for IHC and WB in human, mouse and rat (catalog applications and reactivity). Its IHC image shows paraffin-embedded human breast carcinoma tissue at 1:100; the fixative is unreported (catalog image caption).
Which to pick: For tissue IHC, choose A08306-1: it is a rabbit polyclonal antibody listed for IHC at 1:50–1:200, with a paraffin-section image at 1:100; the fixative is unreported (catalog; catalog image caption). No SKU in this payload is listed for IF/ICC, so there is no IF/ICC pick here (catalog applications). For mouse or rat IHC, A08306-1 has listed reactivity, but its supplied IHC image shows human tissue only (catalog reactivity; catalog image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P55268 (LAMB2_HUMAN, Laminin subunit beta-2).
- Human Protein Atlas. LAMB2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. LAMB2 subcellular location (ICC-IF): Mainly localized to vesicles. In addition localized to the cytosol..
- Human Protein Atlas. LAMB2 antibody validation summary (4 antibodies).
- LAMB2 p.E991K Mutation-Mediated Atherosclerosis in Rabbit. Genes 2026 — PMC13409397.
- LAMB2 novel variant c.2885-9 C>A affects RNA splicing in a minigene assay. Molecular genetics & genomic medicine 2021 — PMC8372075.
- Mutations in LAMB2 Are Associated With Albuminuria and Optic Nerve Hypoplasia With Hypopituitarism. The Journal of clinical endocrinology and metabolism 2020 — PMC7048679.
- PubMed PMID:7698745 — UniProt-cited evidence.
- PubMed PMID:7795887 — UniProt-cited evidence.
- PubMed PMID:19159218 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LAMB2 (Laminin subunit beta-2). Expected localisation ECM and basement membrane; cytoplasmic tissue staining (UniProt; HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A08306-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Adipocytes); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A08306-1); 2. Fixation: Image fixative and duration unreported (datasheet A08306-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-LAMB2, 1:50-1:200 (datasheet A08306-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Secreted ECM deposition may differ from cellular staining (UniProt); Isoform / epitope: No isoforms annotated; mature chain 33–1798 has no cytoplasmic epitope (UniProt); Regulation: Expression regulators unreported (UniProt). Evidence: UniProt P55268, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to vesicles. In addition localized to the cytosol., Datasheet A08306-1, PMC13409397, PMC7048679. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LAMB2/immunohistochemistry-ihc-guide.