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MAP1LC3B / Microtubule-associated protein 1 light chain 3 beta · IHC design guide
Design Immunohistochemistry for MAP1LC3B
Plan chromogenic IHC for MAP1LC3B in paraffin sections using the catalog antibody’s validated conditions (datasheet A01524-2). Compare cytoplasmic staining with the tissue profile, while accounting for the antibody specificity warning (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MAP1LC3B 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 staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in neuronal and glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01524-2) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Appendix+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody may detect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | No isoforms annotated; residues 121–125 are cleaved (UniProt) |
Section 1
Recommended MAP1LC3B IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by 4 published MAP1LC3B IHC protocols spanning gastric, breast, and xenograft tumor tissue, plus a tissue staining review (PMC10883967; PMC8615094; PMC5045373; PMC5617963).
| Sample | Paraffin-embedded human glioma tissue; fixative not specified (datasheet A01524-2) |
| Fixation | Image fixative and duration unreported (datasheet A01524-2); 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 A01524-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01524-2) |
| Primary antibody | Rabbit anti-MAP1LC3B, 1:50 recommended; image 1:100 (datasheet A01524-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01524-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01524-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MAP1LC3B-positive staining in neuronal cells of caudate (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with the catalog antibody’s heat-mediated EDTA pH 8.0 retrieval (datasheet: A01524-2); optimize against the published retrieval conditions for the chosen tissue (PMC5045373; PMC8615094; PMC5617963).
Section 2
What Is the Expected MAP1LC3B Staining Pattern?
MAP1LC3B is associated with cytoplasmic vesicles and autophagosome membranes; it has no transmembrane segment (UniProt Q9GZQ8). In paraffin-section IHC, expect cytoplasmic staining in selected cell populations, including caudate neuronal cells and thyroid glandular cells, both scored High by HPA (HPA: tissue IHC). Interpret these patterns cautiously: HPA rates its tissue IHC reliability Uncertain because the antibodies may target proteins from more than one gene (HPA: tissue IHC reliability).
| Cytoplasmic staining in caudate neuronal cells or thyroid glandular cells, with clearer signal in the listed positive cells than nearby unstained areas (HPA: High in both cell populations). | This agrees with HPA tissue IHC and with MAP1LC3B's cytoplasmic vesicle and autophagosome association (HPA: tissue IHC; UniProt Q9GZQ8 localization). It is a plausible positive pattern, but HPA's Uncertain reliability prevents attribution to MAP1LC3B alone (HPA: tissue IHC reliability). |
| Predominantly nuclear or extracellular chromogen, without a discernible cytoplasmic component (UniProt Q9GZQ8 localization). | That compartment conflicts with the supplied MAP1LC3B localization evidence and warrants an artefact check (UniProt Q9GZQ8 localization). Compare the slide with a control processed in parallel and inspect counterstain, precipitate, and tissue edges (general IHC practice). |
| Strong staining in a cell population listed as Not detected, such as appendix glandular cells or colon endothelial cells (HPA: tissue IHC). | Investigate antibody cross-reactivity or endogenous detection activity before calling it MAP1LC3B (HPA: Uncertain IHC reliability; general IHC practice). An HPA Not detected call is specific to the listed cells and assay; it does not prove that an entire organ lacks protein (HPA: tissue IHC). |
| Broad, similar chromogen across cells and tissue structures, obscuring cytoplasmic detail (HPA: cytoplasmic tissue profile). | This is poor evidence for a cell-specific positive result (HPA: tissue IHC profile). Check a control without primary antibody, blocking, reagent concentration, washing, and development time to distinguish detection background from interpretable staining (general IHC practice). |
| No cytoplasmic signal in caudate neuronal cells or thyroid glandular cells (HPA: High in those cells). | First check section quality and the staining run with a suitable control (general IHC practice). HPA's positive examples make these useful comparison tissues, but its Uncertain reliability means a negative result cannot by itself establish loss of MAP1LC3B (HPA: tissue IHC reliability). |
💡Expected MAP1LC3B appearanceCall a result plausible when cytoplasmic staining is evident in caudate neuronal cells or thyroid glandular cells, where HPA reports High staining; predominantly nuclear or extracellular signal is suspect given UniProt localization, and every assignment remains provisional under HPA's Uncertain IHC reliability (HPA: tissue IHC; UniProt Q9GZQ8 localization).
| Subcellular distribution (UniProt Q9GZQ8 localization). | LC3-II binds autophagic membranes, while the record also places MAP1LC3B in the cytoplasm and endomembrane system (UniProt Q9GZQ8). Thus a cytoplasmic IHC pattern is biologically coherent; a punctate appearance may be informative, but puncta alone do not establish autophagic flux (UniProt Q9GZQ8 function; general IHC interpretation). |
| Cell selection and apparent tissue differences (HPA: tissue IHC; UniProt Q9GZQ8 tissue specificity). | HPA scores caudate neuronal and thyroid glandular cells High, yet heart cardiomyocytes Not detected; UniProt describes abundant expression in heart (HPA: tissue IHC; UniProt Q9GZQ8 tissue specificity). Keep those observations separate when choosing controls; neither source resolves the discrepancy (HPA: Uncertain IHC reliability). |
| Antibody validation and processing (HPA: antibody validation; UniProt Q9GZQ8 processing). | The four listed HPA IHC antibodies have Uncertain validation, and the tissue profile warns of recognition of proteins from multiple genes (HPA: antibodies; HPA: tissue IHC reliability). UniProt records a 1–120 chain and 121–125 propeptide; epitope position for an unspecified antibody is unknown here, so processing effects cannot be predicted (UniProt Q9GZQ8 processing). |
| IF/ICC Q&A: What pattern should a separate IF experiment assess? (HPA: subcellular ICC-IF). | Look primarily for vesicles; HPA additionally reports cytosol and basal body, both with uncertain localization, and cautions that its antibodies may target proteins from multiple genes (HPA: subcellular ICC-IF). This ICC-IF observation does not establish the appearance of chromogenic paraffin-section IHC (HPA: subcellular ICC-IF; HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| A positive reference section shows no interpretable cytoplasmic staining (HPA: High in caudate neurons and thyroid glandular cells). | A failed staining run, unsuitable antibody conditions, or an unsuitable section is possible (general IHC practice). | Review section integrity and run controls; check the catalog antibody's IHC-P instructions for its validated conditions before adjusting the assay (general IHC practice). |
| Chromogen appears mainly nuclear despite expected cytoplasmic distribution (UniProt Q9GZQ8 localization). | Non-specific signal, precipitate, or counterstain confusion may distort compartment assignment (general IHC practice). | Inspect morphology at higher magnification, compare a control without primary antibody, and score cytoplasmic signal separately (general IHC practice). |
| Appendix glandular cells stain strongly despite HPA's Not detected call (HPA: tissue IHC). | Cross-reactivity is plausible given HPA's multi-gene targeting warning; endogenous detection activity is another assay-level possibility (HPA: tissue IHC reliability; general IHC practice). | Compare an independent antibody when available and include a control without primary antibody and an appropriate detection control (general IHC practice). |
| Diffuse background masks differences between cell populations (HPA: cytoplasmic tissue profile). | Incomplete blocking or washing, excessive reagent concentration, or overdevelopment can raise general background (general IHC practice). | Check controls without primary antibody, optimize blocking and washing, and follow the validated IHC-P detection conditions (general IHC practice). |
| Heart cardiomyocytes are negative despite UniProt's high heart expression statement (HPA: Not detected in cardiomyocytes; UniProt Q9GZQ8 tissue specificity). | The supplied sources disagree at different levels of observation; the record does not identify an assay or fixation cause (HPA: tissue IHC; UniProt Q9GZQ8 tissue specificity). | Report the cardiomyocyte observation as measured, verify run controls, and avoid treating heart as an assured positive IHC control (HPA: Uncertain IHC reliability; general IHC practice). |
| Many small cytoplasmic dots are visible and are being scored as increased autophagy (UniProt Q9GZQ8 localization). | Membrane-associated LC3-II can produce a vesicular pattern, but a static stain does not measure formation and clearance rates (UniProt Q9GZQ8 localization; general autophagy interpretation). | Record puncta and diffuse cytoplasmic staining separately; limit the IHC conclusion to observed localization and seek an independent flux assay for a flux claim (general autophagy interpretation). |
Experiment evidence
IHC & IF images showing MAP1LC3B staining
A01524-2 · IHCIHC analysis of LC3B/MAP1LC3B using anti-LC3B/MAP1LC3B antibody (A01524-2). LC3B/MAP1LC3B was detected in a paraffin-embedded section of hum
A01524-2 · IHCIHC analysis of LC3B/MAP1LC3B using anti-LC3B/MAP1LC3B antibody (A01524-2). LC3B/MAP1LC3B was detected in a paraffin-embedded section of mou
M01524 · IHCIHC analysis of LC3B using anti-LC3B antibody (M01524). LC3B was detected in a paraffin-embedded section of human glioma tissue. Heat mediat
M01524 · IHCIHC analysis of LC3B using anti-LC3B antibody (M01524). LC3B was detected in a paraffin-embedded section of mouse brain tissue. Heat mediate
M01524 · IFImmunofluorescent analysis using the Antibody at 1:150 dilution.
M01524-2 · IHCMouse brain was stained with anti-LC3B rabbit antibody
M01524-2 · IHCRat brain was stained with anti-LC3B rabbit antibody
A01524 · IHCImmunohistochemistry of MAP1LC3B in human brain tissue with MAP1LC3B antibody at 5 μg/mL.
A01524 · IFImmunofluorescence of MAP1LC3B in human brain tissue with MAP1LC3B antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000140941-MAP1LC3B/tissue/cerebral+cortex#imid_18806628
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000140941-MAP1LC3B/tissue/colon#imid_18806554
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000140941-MAP1LC3B/tissue/liver#imid_18806563
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000140941-MAP1LC3B/tissue/kidney#imid_18806556
HPA · ICC-IF SK-MEL-30Human Protein Atlas immunofluorescence, SK-MEL-30, antibody HPA042824; green = MAP1LC3B, blue = nucleus, red = microtubules.
HPA · ICC-IF SK-MEL-30Human Protein Atlas immunofluorescence, SK-MEL-30, antibody HPA042824; green = MAP1LC3B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MAP1LC3B IHC & IF
🧪Run caudate first; neuronal cells should stain (HPA: caudate neuronal cells High). Use appendix glandular cells as the negative tissue (HPA: Not detected); on the caudate slide, cells without neuronal staining should lack specific DAB signal, but their target-negative status is unverified (HPA: caudate neuronal cells High).
Positive control tissue: Caudate (Neuronal cells, HPA High)
Negative control tissue: Appendix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MAP1LC3B in SK-MEL-30, SiHa, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Vesicles (supported) (HPA subcellular).
Technical controls: Include a no-primary secondary-only control, a concentration-matched rabbit IgG isotype control, and MAP1LC3B-knockout tissue or cells as a biological negative (selected-SKU caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and inspect caudate neuronal pigment before interpreting brown DAB deposits (selected-SKU caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption). The caption demonstrates heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required; comparative ease of frozen-section IHC or IF is unreported (selected-SKU caption; HPA: ICC-IF images available). Neuronal pigment can complicate DAB interpretation in caudate, so compare its appearance with the no-primary slide (standard IHC practice).
HPA tissue IHC evidence for MAP1LC3B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Caution, targets protein from more than one gene. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Colon | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MAP1LC3B IHC Tips
Use the documented paraffin section workflow as the starting point, then assess cytoplasmic staining and puncta with controls that account for antibody specificity.
Which retrieval conditions should I try first for MAP1LC3B staining in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A01524-2). Apply the same retrieval consistently across comparison sections, then follow the documented 1:100 primary antibody incubation overnight at 4°C when establishing the assay (datasheet A01524-2). If staining is weak, adjust heating time in small increments and inspect morphology before considering a different retrieval buffer (standard IHC practice). Compare each change against a section processed with the original conditions, because stronger diffuse cytoplasmic color alone does not establish better detection of MAP1LC3B associated vesicles (UniProt Q9GZQ8 subcellular location; standard IHC practice).
How should I troubleshoot weak MAP1LC3B staining when fixation history is uncertain?
The documented image uses a paraffin embedded human glioma section but does not report its fixative, so target specific fixation sensitivity is unknown (datasheet A01524-2). Record the available fixation history and compare sections with similar processing before attributing weak staining to antigen loss (standard IHC practice). Run the documented EDTA pH 8.0 retrieval and 1:100 overnight primary incubation on a known staining control alongside the problem section (datasheet A01524-2). If that control stains while the problem section remains weak, examine tissue preservation, section adhesion, and detection performance before changing antibody concentration; those checks cannot by themselves identify a MAP1LC3B specific fixation effect (standard IHC practice).
What staining pattern is plausible for MAP1LC3B in chromogenic tissue IHC?
Expect cytoplasmic staining with possible discrete vesicular puncta, since MAP1LC3B is associated with autophagosome membranes and cytoplasmic vesicles (UniProt Q9GZQ8 subcellular location; HPA: cytoplasmic expression in several tissues). Score diffuse cytoplasmic color and puncta separately when resolution permits, and use adjacent morphology to identify the stained cell population (standard IHC practice). The HPA subcellular assessment supports vesicles but cautions that its antibodies target proteins from multiple genes (HPA: subcellular summary). A nuclear only pattern, or uniform color covering tissue spaces, warrants review of controls and chromogen distribution before it is called MAP1LC3B localisation (UniProt Q9GZQ8 subcellular location; standard IHC practice).
Can this antibody distinguish lipidated LC3-II or other LC3 family proteins in sections?
Do not infer LC3-II specificity from punctate chromogenic staining: the supplied record places LC3-II on autophagic membranes, but the antibody epitope and form selectivity are unspecified (UniProt Q9GZQ8 subcellular location; datasheet A01524-2). MAP1LC3B has a processed chain spanning residues 1–120 and a 121–125 propeptide, with no annotated alternative isoforms in this record (UniProt Q9GZQ8 processing and isoforms). Ask for epitope and cross reactivity information before treating staining as gene specific, especially because the HPA tissue assessment warns about antibodies recognizing proteins from more than one gene (HPA: reliability uncertain). Interpret puncta as a localisation observation rather than direct proof of lipidation (standard IHC practice).
How can IF help check cell identity and puncta seen by chromogenic IHC?
On a separate IF section, multiplex MAP1LC3B with a marker for the cell population being evaluated, selecting that marker from the section’s morphology and experimental question (standard IF practice). Choose fluorophores after inspecting unstained tissue autofluorescence, and place the signal of greatest interest in a channel with lower background (standard IF practice). Use a mild, controlled permeabilisation step to expose cytoplasmic antigen while preserving puncta; MAP1LC3B has no transmembrane segment, but the supplied record does not establish this antibody’s epitope orientation (UniProt Q9GZQ8 topology; standard IF practice). Include single stain and secondary only controls to check bleed through and nonspecific fluorescence before comparing IF puncta with DAB deposits (standard IF practice).
How do I reduce diffuse brown background without losing MAP1LC3B puncta?
First compare a no primary control with the stained section to identify background from detection reagents or endogenous peroxidase (standard IHC practice). The documented tissue workflow used 10% goat serum blocking, 1:100 primary antibody overnight at 4°C, and DAB detection (datasheet A01524-2). If background persists, verify the peroxidase block, wash thoroughly, and shorten chromogen development while keeping a staining control in the same run (standard IHC practice). Adjust primary concentration only after these checks, because diffuse cytoplasmic MAP1LC3B staining is plausible and eliminating all diffuse color could discard signal (HPA: cytoplasmic expression in several tissues; standard IHC practice).
How should I score MAP1LC3B IHC across sections with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring, then keep retrieval, antibody incubation, imaging, and DAB development consistent across sections (standard IHC practice). Report the percentage of positive cells and an H-score for cytoplasmic intensity; when puncta are resolvable, report puncta density per mm² of viable tissue or per counted target cell separately (standard IHC practice). Normalise counts to the number of evaluable cells in the named population and exclude necrotic or folded regions using preset rules (standard IHC practice). Document cell composition because reported staining varies among tissue cell types, and treat HPA comparisons cautiously given its uncertain antibody specificity (HPA: tissue IHC profile and reliability).
When should apparent MAP1LC3B positive cells be treated as artefacts?
A credible result has cytoplasmic or vesicular staining in identifiable cells, consistent with reported MAP1LC3B localisation (UniProt Q9GZQ8 subcellular location; HPA: cytoplasmic tissue profile). Recheck staining confined to cut edges, folds, necrotic areas, or tissue spaces, and compare it with a no primary control for endogenous enzyme or reagent signal (standard IHC practice). Do not assign a brown deposit to a target cell until its nucleus and boundaries can be identified on the counterstained section (standard IHC practice). Finally, neither stronger puncta nor a higher positive cell fraction alone measures autophagic flux, and the HPA tissue reference has uncertain gene specificity (UniProt Q9GZQ8 function; HPA: reliability uncertain).
Boster reagents
Best MAP1LC3B / Microtubule-associated protein 1 light chain 3 beta IHC Antibodies
Anti-MAP1LC3B antibodies have IHC images from human tissue and mouse and rat brain, plus IF images from human brain tissue and HeLa cells (catalog image captions).
Real IHC data
Anti-LC3B/MAP1LC3B Antibody
Cat # A01524-2
Real IHC data
Anti-LC3B MAP1LC3B Rabbit Monoclonal Antibody
Cat # M01524
Real IHC data
Anti-LC3B Rabbit Monoclonal Antibody
Cat # M01524-2
Real IHC data
Anti-MAP1LC3B Antibody
Cat # A01524
A01524 has IHC and IF images from human brain tissue; A01524-2 has paraffin-section IHC images from human glioma and mouse and rat brain (catalog image captions). M01524 has paraffin-section IHC images from human glioma and mouse and rat brain, plus IF images including HeLa cells; M01524-2 has IHC images from human, mouse and rat brain (catalog image captions).
Which to pick: For tissue IHC, choose A01524-2 when a documented paraffin-section workflow matters: its images describe EDTA retrieval at pH 8.0 and primary antibody at 1:100 overnight at 4°C; the fixative is unreported (A01524-2 IHC captions). For IF/ICC, M01524 lists both applications and has an IF image of HeLa cells, although IHC is absent from its application list despite its paraffin-section images (M01524 applications and image captions). For IHC across species, A01524-2 has separate paraffin-section images from human glioma and mouse and rat brain; the fixative is unreported (A01524-2 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9GZQ8 (MLP3B_HUMAN, Microtubule-associated protein 1 light chain 3 beta).
- Human Protein Atlas. MAP1LC3B tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. MAP1LC3B subcellular location (ICC-IF): Mainly localized to vesicles. In addition localized to the cytosol and basal body. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. MAP1LC3B antibody validation summary (5 antibodies).
- Identifying CTH and MAP1LC3B as ferroptosis biomarkers for prognostic indication in gastric cancer decoding. Scientific reports 2024 — PMC10883967.
- Standard Immunohistochemical Assays to Assess Autophagy in Mammalian Tissue. Cells 2017 — PMC5617963.
- Autophagy-related cell death by pan-histone deacetylase inhibition in liver cancer. Oncotarget 2016 — PMC5045373.
- Combined Evaluation of MAP1LC3B and SQSTM1 for Biological and Clinical Significance in Ductal Carcinoma of Breast Cancer. Biomedicines 2021 — PMC8615094.
- PubMed PMID:12932674 — UniProt-cited evidence.
- PubMed PMID:14713272 — UniProt-cited evidence.
- PubMed PMID:12740394 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MAP1LC3B (Microtubule-associated protein 1 light chain 3 beta). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01524-2). Validated IHC antibody: A01524-2 at 1:50 recommended; image 1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Neuronal cells); negative Appendix. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human glioma tissue; fixative not specified (datasheet A01524-2); 2. Fixation: Image fixative and duration unreported (datasheet A01524-2); 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 A01524-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01524-2); 8. Primary antibody: Rabbit anti-MAP1LC3B, 1:50 recommended; image 1:100 (datasheet A01524-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01524-2). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Antibody may detect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: No isoforms annotated; residues 121–125 are cleaved (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt Q9GZQ8, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to vesicles. In addition localized to the cytosol…, Datasheet A01524-2, PMC10883967, PMC5617963. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MAP1LC3B/immunohistochemistry-ihc-guide.