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LALBA / Alpha-lactalbumin · IHC design guide
Design Immunohistochemistry for LALBA
Plan LALBA IHC in lactating breast, where glandular and ductal cytoplasmic staining is reported (HPA tissue IHC). This guide covers paraffin-section handling and interpretation of plasma staining in light of LALBA secretion (datasheet A04858; HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LALBA 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 lactating breast (HPA tissue IHC) | |
| Staining pattern | Glandular and ductal cytoplasm; additional plasma staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A04858) | |
| Positive control | Lactating breast | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Use formalin-fixed paraffin sections (selected-SKU IHC image A04858); keep fixation consistent. (standard IHC practice; not target-specific) | |
| Caveat | Secreted LALBA may stain plasma beyond producer cells (UniProt; HPA tissue IHC) | |
| Regulation | Mammary gland specific; secreted in milk (UniProt) | |
| Isoform / epitope | No reported isoforms; mature chain spans residues 20–142 (UniProt) |
Section 1
Recommended LALBA IHC & IF Protocols
The catalog antibody protocol uses citrate pH 6 retrieval (datasheet: A04858). Two published IHC protocols provide breast cancer and mare mammary gland examples (PMC12898300; PMC11350905).
| Sample | FFPE human breast carcinoma tissue (datasheet A04858) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A04858); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6 (datasheet A04858); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-LALBA, 1:25 (datasheet A04858) |
| Primary incubation | 1 hours at 37°C (datasheet A04858) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LALBA-positive staining in ductal cells of lactating breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in glandular and ductal cells in lactating breast. Additional plasma positivity. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6 HIER for the catalog antibody (datasheet: A04858). The published breast cancer protocol used EDTA pH 8 with a different antibody (PMC12898300: methods).
Section 2
What Is the Expected LALBA Staining Pattern?
LALBA is a secreted, non-transmembrane protein associated with milk production (UniProt P00709). In paraffin-section IHC, expect cytoplasmic staining of glandular and ductal cells in lactating breast, with additional plasma positivity (HPA tissue IHC). HPA rates the tissue pattern Enhanced, while noting that a secreted protein’s tissue RNA and protein locations may differ (HPA tissue IHC).
| Strong cytoplasmic staining in lactating-breast ductal cells; glandular cells and plasma may also stain (HPA tissue IHC). | This matches the reported LALBA pattern (HPA tissue IHC). Assess the cellular cytoplasm separately from plasma positivity: secretion can place protein beyond the cell that produced it (UniProt P00709; HPA tissue IHC). |
| Predominantly nuclear or sharply membrane-bound staining in otherwise positive ductal cells. | Treat this as a localisation mismatch requiring review, since HPA reports cytoplasmic staining and UniProt describes secretion without a transmembrane segment (HPA tissue IHC; UniProt P00709). Check morphology, counterstain and controls before assigning biological meaning (standard IHC practice). |
| Prominent staining of adipocytes in breast or adipose tissue, or of unrelated cells reported as negative. | These cells are listed as not detected in the supplied HPA tissue data (HPA tissue IHC). Consider antibody cross-reactivity or endogenous detection activity; use a negative tissue and detection-only control to distinguish them (standard IHC practice). |
| Widespread, poorly localised colour across cells, stroma or slide background. | Diffuse colour alone cannot establish LALBA expression when the reference pattern is cell-associated cytoplasm with additional plasma positivity (HPA tissue IHC). Review detection-only and negative-tissue controls for nonspecific deposition or background (standard IHC practice). |
| No convincing ductal-cell signal in a lactating-breast section. | This conflicts with the High ductal-cell level reported for lactating breast (HPA tissue IHC). First verify that the section contains appropriate ducts, then review antibody and detection controls; a negative result alone does not identify the failed step (standard IHC practice). |
💡Expected LALBA appearanceCall the result positive when lactating-breast ductal or glandular cytoplasm stains convincingly, especially with High ductal signal; isolated nuclear, membrane-bound or adipocyte staining is suspect (HPA tissue IHC; UniProt P00709).
| Positive and negative tissue context | Lactating-breast ductal cells provide a reported High reference; adipocytes in breast and adipose tissue are not detected (HPA tissue IHC). Compare these compartments within the relevant morphology before scoring (standard IHC practice). |
| Secretion and cellular location | LALBA has a signal peptide at residues 1–19 and a mature chain at 20–142; UniProt lists it as secreted with no transmembrane segment (UniProt P00709). Interpret plasma positivity alongside the cytoplasmic tissue pattern (HPA tissue IHC). |
| Protein processing and glycosylation | UniProt lists glycosylation sites at residues 64 and 90 and no propeptide or reported isoforms (UniProt P00709). These annotations describe the protein but do not establish an antibody epitope or a target-specific retrieval requirement. |
| Evidence behind the IHC pattern | HPA rates the tissue result Enhanced; two listed rabbit polyclonal antibodies have Enhanced IHC validation, and one listed rabbit monoclonal antibody is Supported (HPA tissue IHC; HPA antibodies). HPA also says no paired tissue staining or RNA data are available for comparison (HPA tissue IHC). |
| Fixation and antigen retrieval | Target-specific fixation sensitivity and retrieval conditions are unreported in the supplied UniProt and HPA records. If staining fails, assess retrieval within a controlled IHC optimisation using the same known-positive tissue; do not infer a LALBA-specific effect from topology or glycosylation (standard IHC practice; UniProt P00709). |
| IF/ICC: what localisation should be expected? | HPA lists LALBA as secreted but supplies no main ICC-IF location, cell-line images or ICC validation for the listed antibodies (HPA subcellular; HPA antibodies). Use the separate IF/ICC guide for assay design; the IHC tissue pattern alone cannot define an ICC-IF image. |
| Situation | Likely cause | Next action |
|---|---|---|
| Lactating-breast ductal cells appear negative. | The expected High ductal reference is absent (HPA tissue IHC); an unsuitable section or assay failure remains possible (standard IHC practice). | Confirm lactating-breast ductal morphology and inspect positive and detection controls; optimise retrieval or antibody conditions only against those controls (standard IHC practice). |
| Adipocytes stain as strongly as ductal cells. | Breast adipocytes are reported not detected, so this result is discordant with the reference pattern (HPA tissue IHC). | Compare adipocytes with lactating-breast ducts and a negative tissue; check detection-only staining before attributing colour to LALBA (standard IHC practice). |
| Nuclei dominate the signal. | Nuclear staining does not match HPA’s cytoplasmic description or UniProt’s secreted localisation (HPA tissue IHC; UniProt P00709). | Recheck focus, counterstain and compartment boundaries, then compare antibody-stained and detection-only sections (standard IHC practice). |
| Colour is diffuse or obscures cell boundaries. | Nonspecific chromogen deposition or detection background can make the reported cellular pattern unreadable (standard IHC practice; HPA tissue IHC). | Inspect negative-tissue and detection-only controls; review blocking, detection and wash conditions before scoring compartments (standard IHC practice). |
| Plasma is positive but cellular signal is weak. | Additional plasma positivity is reported, and secreted protein can be found away from producer cells (HPA tissue IHC; UniProt P00709). | Score plasma and cellular cytoplasm separately; confirm ductal morphology and compare with a known-positive section before calling ductal cells negative (standard IHC practice). |
| An IF/ICC image is assigned a precise organelle pattern. | HPA provides no main ICC-IF location, images or ICC validation for the listed antibodies (HPA subcellular; HPA antibodies). | Record the observed distribution without claiming HPA-supported organelle localisation; use assay-specific controls and the separate IF/ICC guide for interpretation (standard IF practice). |
Experiment evidence
IHC & IF images showing LALBA staining
A04858 · IHCLALBA Antibody (Center) (A04858)immunohistochemistry analysis in formalin fixed and paraffin embedded human breast carcinoma followed by per
A04858 · IHCA04858 staining LALBA in Human breast tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). T
M04858 · IHCImmunohistochemical analysis of paraffin-embedded Human-liver tissue. 1, alpha Lactalbumin Monoclonal Antibody (9E9) was diluted at 1:200 (4
M04858 · IFImmunofluorescence analysis of Human-breast tissue. 1, alpha Lactalbumin Monoclonal Antibody (9E9) (red) was diluted at 1:200 (4°C, overnigh
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000167531-LALBA/tissue/cerebral+cortex#imid_9522848
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000167531-LALBA/tissue/colon#imid_9522774
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000167531-LALBA/tissue/liver#imid_9522767
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000167531-LALBA/tissue/kidney#imid_9522776
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LALBA IHC & IF
🧪Run lactating breast first and require staining in ductal cells (High; HPA: lactating breast). Use breast adipocytes as a negative comparison (Not detected; HPA: breast); adipocytes present on the positive slide should lack specific cellular staining (HPA: breast adipocytes).
Positive control tissue: Lactating breast (Ductal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for LALBA; derive a cell-line control from the positive tissue's cell type (Ductal cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and host-matched immunoglobulin controls, using an isotype-matched control for a monoclonal antibody or nonimmune IgG for a polyclonal antibody (standard IHC control practice). Include a LALBA knockout specimen or immunizing-peptide competition where available; block endogenous peroxidase for DAB detection and assess endogenous biotin if using avidin-based detection (standard IHC control practice; selected tissue-IHC caption: peroxidase-conjugated secondary and DAB).
⚠️Feasibility: The selected tissue-IHC caption reports formalin-fixed, paraffin-embedded human breast carcinoma, but gives no target-specific fixation window (selected tissue-IHC caption). Retrieval dependence is unreported, and the supplied evidence does not establish whether frozen sections or IF are easier; HPA lists no ICC-IF cell-line images (HPA: subcellular record). Because LALBA is secreted in milk, luminal or extracellular signal may complicate cellular scoring in lactating breast (UniProt P00709: secreted in milk; HPA: lactating breast ductal cells).
HPA tissue IHC evidence for LALBA
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — No tissue staining or RNA expression data available for comparison. Secreted protein, tissue location of RNA and protein is expected to differ.). 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 |
|---|---|---|---|---|
| Lactating breast | Ductal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LALBA IHC Tips
Troubleshoot LALBA staining in paraffin sections by checking retrieval, controls, cellular localisation and detection artefacts before comparing staining scores.
Which retrieval should I start with for LALBA in paraffin sections?
Start with heat-mediated citrate retrieval at pH 6 (datasheet A04858). Process positive and negative control sections alongside the study sections, and keep heating and cooling conditions consistent across the batch (standard IHC practice). If staining is weak, compare a longer heating interval with the starting condition in a small control series, while checking for tissue damage or increased background (standard IHC practice). LALBA is secreted and has a signal peptide spanning residues 1–19 (UniProt P00709), so judge retrieval by staining in expected cells and material, rather than requiring a membrane outline (HPA tissue IHC; UniProt P00709).
How should I troubleshoot variable LALBA staining after fixation?
The product image shows staining in formalin-fixed, paraffin-embedded human breast carcinoma (A04858 tissue-IHC caption), but LALBA-specific sensitivity to fixation duration is unknown. Record fixation conditions, section thickness and storage history for each case before comparing staining intensity (standard IHC practice). When signal varies, stain matched control sections in the same run and change one processing variable at a time (standard IHC practice). Do not infer fixation tolerance from lactating-breast staining patterns (HPA tissue IHC) or from LALBA secretion, glycosylation or disulfide bonds (UniProt P00709); none of those observations measures the effect of fixation on this antibody.
Where should convincing LALBA staining appear?
Look first for cytoplasmic staining in glandular and ductal cells of lactating breast; extracellular or plasma positivity can also occur (HPA tissue IHC). LALBA has no transmembrane segment and is secreted into milk (UniProt P00709), so a continuous cell-surface rim is not the expected defining pattern. Compare cell-associated staining with adjacent secreted material and assess each compartment separately (HPA tissue IHC; standard IHC practice). If only an isolated rim or diffuse staining appears, review the negative reagent control, counterstain and tissue morphology before assigning it to LALBA (standard IHC practice).
Could LALBA processing or glycosylation explain discordant antibody staining?
LALBA has no listed isoforms; its 142-residue precursor loses a signal peptide at residues 1–19, leaving the 20–142 chain (UniProt P00709). Glycosylation sites are listed at residues 64 and 90 (UniProt P00709), but these annotations do not establish whether either modification affects this antibody. The product is described as a center-directed antibody without a precise epitope sequence in the supplied caption (A04858 tissue-IHC caption). If another antibody gives a different pattern, compare their documented immunogens and controls, then test matched sections under the same retrieval and detection conditions (standard IHC practice).
How can I check an IF signal against the chromogenic LALBA pattern?
Use a marker identifying the expected glandular or ductal cell population for multiplex IF, then compare its distribution with LALBA signal (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and an imaging channel with low tissue autofluorescence, using single-stain and unstained controls to check bleed-through and background (standard IF practice). LALBA is secreted and has no transmembrane segment (UniProt P00709); if the IF antibody recognizes an intracellular epitope, optimise gentle permeabilisation on control sections, while assessing extracellular signal separately (standard IF practice). IF conditions need their own validation: the supplied A04858 image documents chromogenic tissue IHC, not IF performance (A04858 tissue-IHC caption).
What should I check when DAB obscures weak LALBA staining?
Run a no-primary control and review endogenous peroxidase blocking before attributing diffuse DAB deposition to LALBA (standard chromogenic IHC practice). Titrate primary antibody and detection reagent on matched control sections, keeping retrieval and DAB development consistent (standard IHC practice). Inspect section edges, folds and damaged areas separately because these can collect stain or develop unevenly (standard IHC practice). Expected staining is cytoplasmic in lactating-breast glandular and ductal cells, with additional plasma positivity reported (HPA tissue IHC); widespread staining of unrelated cells should prompt a background check before scoring (HPA tissue IHC; standard IHC practice).
How should I score LALBA when secreted material also stains? ⚠ ANSWER MARKED FOR VERIFICATION
Define a cellular scoring region before analysis and record the percentage of positive glandular or ductal cells plus staining intensity, or calculate an H-score from those categories (HPA tissue IHC; standard IHC scoring practice). Normalise cell counts to the number of eligible epithelial cells, and report any extracellular or plasma staining as a separate compartment (HPA tissue IHC; standard IHC scoring practice). If measuring positive-cell density, express it per mm² of analysed tissue and state the excluded regions (standard IHC scoring practice). Apply one intensity threshold and one counterstain-based cell definition across all slides in the comparison (standard IHC scoring practice).
How do I distinguish credible LALBA positivity from artefact?
Credible staining should fit the sampled tissue and the reported cytoplasmic pattern in lactating-breast glandular and ductal cells; plasma positivity may accompany it (HPA tissue IHC). LALBA is a secreted protein without a transmembrane segment (UniProt P00709), so a membrane-only pattern needs additional scrutiny. Check unexpected staining in other cell types against morphology, a no-primary control and a matched positive control (standard IHC practice). Exclude section-edge staining, necrotic regions and signal persisting when the primary antibody is omitted; the latter can reflect endogenous enzyme activity or detection background (standard chromogenic IHC practice).
Boster reagents
Best LALBA / Alpha-lactalbumin IHC Antibodies
Two human-reactive anti-LALBA antibodies have IHC images from breast and liver tissue; one also has an IF image from human breast tissue (catalog reactivity; IHC and IF image captions).
Real IHC data
Anti-LALBA Antibody (Center)
Cat # A04858
Real IHC data
Anti-alpha Lactalbumin LALBA Monoclonal Antibody
Cat # M04858
A04858 will render with an IHC image of formalin-fixed, paraffin-embedded human breast carcinoma (A04858 IHC caption). M04858 will render with an IHC image of paraffin-embedded human liver; its separate IF image shows human breast tissue (M04858 IHC and IF captions).
Which to pick: For breast tissue IHC, A04858 is a rabbit polyclonal with a formalin-fixed, paraffin-embedded breast carcinoma example (catalog host; A04858 IHC caption). For liver tissue IHC or breast tissue IF, M04858 is a mouse monoclonal with those respective images; its liver caption specifies paraffin embedding but does not report a fixative, and ICC appears in its short description without an ICC image (catalog host and short description; M04858 IHC and IF captions). Neither SKU has documented cross-species reactivity: both list human only (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P00709 (LALBA_HUMAN, Alpha-lactalbumin).
- Human Protein Atlas. LALBA tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LALBA subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. LALBA antibody validation summary (3 antibodies).
- Expression Patterns of LALBA and Nucleolin and Their Clinical, Prognostic, and Immune Relevance in Breast Cancer Tissues of Mexican Patients. International journal of molecular sciences 2026 — PMC12898300.
- A Comparative Analysis of the Gene Expression Profiles in the Mammary Glands of Lactating and Nonlactating Mares at the Second Month of Gestation. Animals : an open access journal from MDPI 2024 — PMC11350905.
- Polyherbal formula (ASILACT®) induces Milk production in lactating rats through Upregulation of α-Lactalbumin and aquaporin expression. BMC complementary medicine and therapies 2020 — PMC7690098.
- PubMed PMID:6285305 — UniProt-cited evidence.
- PubMed PMID:2954544 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LALBA (Alpha-lactalbumin). Expected localisation Cytoplasmic in lactating breast (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A04858). Validated IHC antibody: A04858 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Lactating breast (Ductal cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: FFPE human breast carcinoma tissue (datasheet A04858); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A04858); 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: Citrate pH 6 (datasheet A04858); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-LALBA, 1:25 (datasheet A04858); 9. Primary incubation: 1 hours at 37°C (datasheet A04858). Watch for: Fixation: Use formalin-fixed paraffin sections (selected-SKU IHC image A04858); keep fixation consistent. (standard IHC practice; not target-specific); Caveat: Secreted LALBA may stain plasma beyond producer cells (UniProt; HPA tissue IHC); Isoform / epitope: No reported isoforms; mature chain spans residues 20–142 (UniProt); Regulation: Mammary gland specific; secreted in milk (UniProt). Evidence: UniProt P00709, HPA tissue IHC · Enhanced, HPA ICC-IF · Secreted, Datasheet A04858, PMC12898300, PMC11350905. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LALBA/immunohistochemistry-ihc-guide.