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LIF / Leukemia inhibitory factor · IHC design guide
Design Immunohistochemistry for LIF
Plan chromogenic IHC for LIF in paraffin sections using the catalog antibody’s documented conditions (datasheet PB9036). Interpret cytoplasmic staining against HPA’s uncertain tissue profile, accounting for LIF secretion (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LIF 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 tissue staining; LIF is secreted (HPA tissue IHC; UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet PB9036) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Smooth muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet PB9036); verify before use. | |
| Caveat | Secreted LIF may stain away from its producer cells (HPA tissue IHC; UniProt) | |
| Regulation | Regulation not specified in the record (UniProt) | |
| Isoform / epitope | 2 isoforms; signal peptide 1–22 is removed; epitope unreported (UniProt) |
Section 1
Recommended LIF IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by 4 published LIF immunohistochemistry protocols spanning paraffin and frozen sections.
| Sample | Paraffin-embedded human lung cancer tissues; fixative not specified (datasheet PB9036) |
| Fixation | Image fixative and duration unreported (datasheet PB9036); 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, 20 min (datasheet PB9036) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9036) |
| Primary antibody | Rabbit anti-LIF, 0.5-1μg/ml (datasheet PB9036) |
| Primary incubation | Overnight at 4 °C (datasheet PB9036) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9036) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LIF-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteFor paraffin sections with the catalog antibody, start with heat-mediated citrate retrieval at pH 6 (datasheet PB9036); adapt retrieval when following a published antibody-specific protocol.
Section 2
What Is the Expected LIF Staining Pattern?
LIF is a secreted protein with no transmembrane segment (UniProt P15018 topology). In tissue IHC, expect predominantly cytoplasmic staining; HPA reports medium staining in adipocytes, respiratory epithelial cells, neurons and selected glandular cells (HPA tissue IHC). Treat this as a provisional pattern: HPA rates its tissue IHC reliability Uncertain because secretion can separate protein location from RNA expression, and retesting is pending (HPA tissue IHC).
| Cytoplasmic staining in adipocytes or bronchial respiratory epithelial cells, with interpretable cell borders. | This fits HPA's reported medium staining in those cells and its broad cytoplasmic profile (HPA tissue IHC). Score the relevant cells and their intensity; the HPA pattern is provisional because tissue IHC reliability is Uncertain (HPA tissue IHC). |
| Strong, sharply nuclear or membrane-restricted staining dominates the section. | That pattern differs from HPA's cytoplasmic tissue profile and supported cytosolic ICC-IF location (HPA tissue IHC; HPA subcellular). Review morphology and controls before assigning it to LIF; secretion alone does not establish a membrane-restricted pattern (UniProt P15018 topology). |
| Smooth muscle cells stain strongly, especially when control sections show similar color. | HPA reports LIF as not detected in smooth muscle cells (HPA tissue IHC). Consider antibody cross-reactivity or endogenous detection activity; inspect controls before calling these cells positive. A single unexpected stain cannot resolve HPA's Uncertain tissue reliability (HPA tissue IHC). |
| Diffuse color covers extracellular space, tissue edges or many cell types without clear boundaries. | The distribution is difficult to score against HPA's cell-associated cytoplasmic observations (HPA tissue IHC). LIF is secreted, so extracellular material is biologically possible, but diffuse chromogen alone cannot identify its source (UniProt P15018 subcellular location; general IHC interpretation). |
| No staining appears in an adipocyte-rich area with preserved morphology. | HPA reports medium adipocyte staining in adipose tissue and breast (HPA tissue IHC). Check that the cells are present, then review assay controls and detection. Because HPA tissue IHC is Uncertain, absence in one section does not by itself establish assay failure (HPA tissue IHC). |
💡Expected LIF appearanceA provisional positive call is cell-associated cytoplasmic staining of about medium intensity in HPA-listed cells, such as adipocytes or bronchial respiratory epithelium; dominant nuclear staining or indiscriminate diffuse color needs control-based investigation (HPA tissue IHC; general IHC interpretation).
| Secretion and processing (UniProt P15018 topology) | LIF has a signal peptide at residues 1–22 and a mature chain at 23–202; it lacks a transmembrane segment (UniProt P15018). Cytoplasmic IHC can reflect cellular protein, while extracellular staining requires cautious interpretation because secreted protein may move beyond its producing cell (UniProt P15018; HPA tissue IHC). |
| Glycosylation and isoforms (UniProt P15018) | UniProt lists six glycosylation sites and two isoforms (UniProt P15018). Without an antibody epitope or isoform-specific validation in this payload, these annotations cannot predict which form the stain detects or justify a particular retrieval condition. |
| Tissue distribution and confidence (HPA tissue IHC) | HPA reports medium staining in eight listed tissue and cell combinations, low staining in six others, and no detection in smooth muscle cells (HPA tissue IHC). Its overall tissue IHC reliability is Uncertain, so these are comparison patterns rather than definitive positive and negative standards (HPA tissue IHC). |
| IHC antibody evidence (HPA antibodies) | The supplied HPA antibody record marks HPA018844 IHC as Uncertain; CAB025985 has no IHC status listed (HPA antibodies). Neither entry supplies an Enhanced IHC validation designation, so agreement with the reported tissue pattern should be interpreted with that limitation. |
| IF/ICC: what localization is expected? (HPA subcellular) | Mainly cytosolic signal is supported in HPA ICC-IF; images are listed for A-431, U-251MG and U2OS, and both supplied antibodies have Supported ICC status (HPA subcellular; HPA antibodies). That ICC evidence does not establish IHC reliability or an IF/ICC protocol (HPA antibodies). |
| Antigen retrieval choice (general IHC practice) | Evaluate retrieval against the IHC-validated antibody's instructions and assay controls (general IHC practice). Neither UniProt nor the supplied HPA records reports LIF-specific fixation sensitivity or an optimal retrieval condition; HPA staining levels cannot establish either effect (UniProt P15018; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal appears in the test section or an HPA-listed medium-staining tissue. | The sampled cells may be absent, or the IHC workflow may have failed; HPA's medium calls are provisional (HPA tissue IHC; general IHC practice). | Confirm the expected cell type and morphology, then check reagent and detection controls and the antibody's IHC instructions (general IHC practice). Repeat before interpreting a single blank section as biological absence. |
| A low-staining HPA tissue gives a faint, patchy result. | HPA lists several cell types at Low intensity, including caudate glia and duodenal glandular cells (HPA tissue IHC). Faint staining is therefore harder to separate from background. | Compare with a listed medium-staining cell type and the negative control under the same scoring criteria (HPA tissue IHC; general IHC practice). Record ambiguous cells as indeterminate rather than assigning a strong positive call. |
| Nuclei or cell membranes stain more strongly than cytoplasm. | A dominant nuclear or membrane-restricted pattern conflicts with HPA's cytoplasmic tissue profile and supported cytosolic ICC-IF localization (HPA tissue IHC; HPA subcellular). | Inspect counterstain, morphology and detection controls, then reassess antibody specificity (general IHC practice). Do not use the receptor-signaling description to label membrane staining as LIF: LIF itself is secreted (UniProt P15018). |
| Smooth muscle cells stain, despite an otherwise plausible section. | HPA reports smooth muscle cells as Not detected; unexpected color could reflect cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). | Compare smooth muscle with adjacent expected cell types, and check a no-primary control and the detection-system controls (general IHC practice). Treat persistent staining as unresolved until specificity is supported. |
| Broad diffuse chromogen obscures cellular localization. | Background from blocking, detection or washing can obscure cell-associated staining (general IHC practice). LIF secretion also makes extracellular signal possible, but its source cannot be inferred from diffuse color alone (UniProt P15018). | Review negative controls, blocking and wash steps, then score only interpretable cell-associated staining (general IHC practice). Document extracellular color separately from HPA's reported cytoplasmic pattern (HPA tissue IHC). |
| IHC and IF/ICC appear to disagree. | HPA rates tissue IHC Uncertain while reporting supported cytosolic ICC-IF localization; the two observations have different validation status (HPA tissue IHC; HPA subcellular). | Compare compartment and cell identity within each assay, check its controls, and report the discordance with those source-specific confidence levels (HPA tissue IHC; HPA subcellular; general IHC practice). |
Experiment evidence
IHC & IF images showing LIF staining
PB9036 · IHCIHC analysis of LIF using anti-LIF antibody (PB9036). LIF was detected in paraffin-embedded section of human lung cancer tissues. Heat media
PB9036 · IHCIHC analysis of LIF using anti-LIF antibody (PB9036). LIF was detected in paraffin-embedded section of human placenta tissues. Heat mediated
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000128342-LIF/tissue/cerebral+cortex#imid_5004428
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000128342-LIF/tissue/colon#imid_5004362
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000128342-LIF/tissue/liver#imid_5004355
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000128342-LIF/tissue/gallbladder#imid_5004339
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018844; green = LIF, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018844; green = LIF, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LIF IHC & IF
🧪Run adipose tissue first and look for staining in adipocytes (HPA: Medium in adipocytes). Use smooth muscle cells as the negative tissue (HPA: Not detected in smooth muscle cells); on the adipose slide, visibly unstained neighboring cells can show background, but HPA does not identify a verified internal-negative cell type.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Smooth muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LIF in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched irrelevant rabbit IgG control matched to the primary antibody’s clonality (PB9036 caption: rabbit primary). Use LIF knockout tissue or peptide competition as a target-specific negative control; for adipose IHC, block endogenous peroxidase and assess endogenous biotin before using the caption’s biotin-based DAB detection (PB9036 caption: biotinylated secondary, SABC, DAB).
⚠️Feasibility: No supplied evidence reports a target-specific fixation window or fixation effect, and the selected PB9036 paraffin-section caption does not state the fixative (PB9036 caption: fixative unreported). Heat retrieval in citrate buffer at pH 6 for 20 minutes was used, but whether retrieval is required is unreported; the evidence does not establish that frozen sections or IF are easier (PB9036 caption: retrieval conditions). In paraffin adipose sections, lipid extraction can leave clear vacuoles, so score cellular staining rather than the vacuoles (standard IHC practice).
HPA tissue IHC evidence for LIF
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex. Pending retesting.). 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LIF IHC Tips
Use the catalog antibody’s paraffin-section workflow as the IHC starting point, and interpret LIF staining cautiously because secreted protein can appear away from its producing cells (UniProt P15018; HPA tissue IHC).
Which retrieval conditions should I start with when LIF staining is weak?
Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet PB9036). Keep section thickness, heating, cooling, and subsequent incubation conditions consistent while comparing retrieval runs, so changes in staining can be attributed to the retrieval step (standard IHC practice). The selected paraffin-section example used 1 μg/mL primary antibody overnight at 4°C, providing a documented starting concentration after retrieval (datasheet PB9036). If staining remains weak, compare a small retrieval time series alongside the documented condition and check tissue integrity, because excessive heating can damage morphology and make faint signal difficult to assess (standard IHC practice).
How should I troubleshoot suspected fixation effects on LIF IHC?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative (datasheet PB9036). Record the actual fixative, fixation duration, processing history, and section age for each specimen before comparing staining, because these variables can affect antigen preservation and access in IHC (standard IHC practice). Run specimens with different processing histories through the same citrate pH 6, 20-minute retrieval and detection workflow to limit procedural variation (datasheet PB9036; standard IHC practice). If a group stains weakly, compare it with a similarly processed reference section and check morphology and control staining before attributing the difference to LIF abundance (standard IHC practice).
Where should LIF signal appear in a paraffin section?
Assess cellular and extracellular staining separately: LIF is secreted and has no annotated transmembrane segment (UniProt P15018 topology). Cytoplasmic staining is reported in most tissues, although the tissue IHC profile has uncertain reliability (HPA tissue IHC). For a cellular reference, HPA reports medium staining in adipocytes of adipose tissue and respiratory epithelial cells of bronchus, with smooth muscle cells listed as not detected (HPA tissue IHC). When staining surrounds cells, document its distribution without assigning the producing cell, because a secreted protein can be detected away from its site of synthesis (UniProt P15018 secreted; standard IHC interpretation).
Can this stain distinguish LIF isoforms or processing states?
Do not assign an isoform from this stain alone: the record lists 2 isoforms, but the supplied antibody evidence does not define an isoform-specific epitope (UniProt P15018 isoforms; datasheet PB9036). The annotated signal peptide spans residues 1–22, and the mature chain spans 23–202, so epitope position would matter when interpreting recognition of processed LIF (UniProt P15018 processing). LIF also has 6 annotated glycosylation sites, which makes antibody access a reasonable question to investigate without assuming that glycosylation changes this antibody’s staining (UniProt P15018 glycosylation; standard IHC practice). Compare documented epitope information and orthogonal evidence before claiming isoform or processing-state specificity (standard IHC practice).
How can I adapt LIF detection to multiplex immunofluorescence?
Treat IF as a separately optimized application; the selected antibody example documents chromogenic IHC in a paraffin section, not an IF protocol (datasheet PB9036). Pair LIF with a marker identifying the cell population under study, and inspect each channel alone before interpreting overlap, since secreted LIF may lie outside its producing cell (UniProt P15018 secreted; standard IF practice). Measure unstained tissue autofluorescence first, then choose a fluorophore channel with adequate separation from that background and the other marker (standard IF practice). If the antibody recognizes an intracellular epitope, optimize permeabilisation for access to it; an accessible extracellular epitope may require a different approach, and the supplied evidence does not locate this antibody’s epitope (standard IF practice; datasheet PB9036).
What should I check when LIF IHC shows diffuse brown background?
First inspect a section processed without primary antibody to assess detection-system background, then check the peroxidase block and DAB development time (standard chromogenic IHC practice). The selected example used 10% goat serum, a biotinylated secondary antibody, a streptavidin–biotin detection complex, and DAB; these are documented workflow components, not proof that every brown deposit represents LIF (datasheet PB9036). Compare background in tissue edges, damaged areas, and intact interiors, and shorten development or adjust blocking only while preserving the documented reference run (standard IHC practice). Because LIF is secreted, diffuse staining needs spatial and control-based assessment before it is interpreted as extracellular protein (UniProt P15018 secreted; standard IHC interpretation).
How should I quantify LIF staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define comparable intact tissue regions and score cellular staining separately from extracellular deposits, because LIF is secreted (UniProt P15018 secreted; standard IHC practice). For cellular staining, report the percentage of positive cells and an H-score based on intensity categories 0–3; for spatially discrete positive cells, density per mm² can also be informative (standard IHC practice). Normalize counts or stained area to the relevant viable tissue area or eligible cell population, and apply identical thresholds and imaging settings across sections (standard IHC practice). Report cell type and compartment with each result, since the HPA tissue IHC profile is marked uncertain and RNA location need not match protein location (HPA tissue IHC).
How do I distinguish plausible LIF staining from artefact?
Look for reproducible staining in intact cells or plausible extracellular spaces while recording the compartment, because LIF is secreted and HPA reports predominantly cytoplasmic tissue staining with uncertain reliability (UniProt P15018 secreted; HPA tissue IHC). Check whether signal tracks tissue edges or necrotic areas, where staining can reflect section artefact rather than interpretable antigen distribution (standard IHC practice). Compare a no-primary section to identify detection-system signal, including residual endogenous peroxidase activity in a DAB workflow (standard chromogenic IHC practice). Use cell identity and independent evidence before assigning a producer: HPA lists bronchial respiratory epithelial cells as medium and smooth muscle cells as not detected, but its tissue profile remains uncertain (HPA tissue IHC).
Boster reagents
Best LIF / Leukemia inhibitory factor IHC Antibodies
PB9036 has real IHC staining data from human paraffin-embedded lung cancer and placenta sections (catalog image captions). The catalog supplies no IF/ICC data (catalog applications and image captions).
Real IHC data
Anti-LIF Antibody ®
Cat # PB9036
PB9036 is listed for human IHC and WB (catalog applications and reactivity). Its IHC captions show staining of human paraffin-embedded lung cancer and placenta sections (catalog image captions).
Which to pick: Choose PB9036 for human paraffin-section IHC: its stated range is 0.5–1 μg/ml, and its captions use citrate retrieval at pH 6 for 20 minutes (datasheet dilution; PB9036 IHC captions). The captions do not report the fixative, and no clone is specified (PB9036 IHC captions; catalog clone field). No listed SKU is validated here for IF/ICC or cross-species use: PB9036 lists human reactivity and IHC/WB applications only (catalog reactivity and applications).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P15018 (LIF_HUMAN, Leukemia inhibitory factor).
- Human Protein Atlas. LIF tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. LIF subcellular location (ICC-IF): Mainly localized to the cytosol..
- Human Protein Atlas. LIF antibody validation summary (2 antibodies).
- Effects of "Bu Shen Huo Xue Decoction" on the Endometrial Morphology and Expression of Leukaemia Inhibitory Factor in the Rat Uterus during the Oestrous Cycle. Evidence-based complementary and alternative medicine : eCAM 2013 — PMC3655613.
- Investigation of the effects of recombinant and urinary FSH used for ovulation induction on pregnancy results in an experimental animal model in terms of endometrial receptivity markers. BMC pregnancy and childbirth 2026 — PMC12911375.
- Immunohistochemical expression pattern of leukemia inhibitory factor and its receptor may mark a tipping point in the progression of oral cancer. Scientific reports 2025 — PMC12508188.
- Expression of leukemia inhibitory factor (LIF) and its receptor gp190 in human liver and in cultured human liver myofibroblasts. Cloning of new isoforms of LIF mRNA. Comparative hepatology 2004 — PMC538256.
- PubMed PMID:3143918 — UniProt-cited evidence.
- PubMed PMID:2475312 — UniProt-cited evidence.
- PubMed PMID:1692837 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LIF (Leukemia inhibitory factor). Expected localisation Cytoplasmic tissue staining; LIF is secreted (HPA tissue IHC; UniProt). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet PB9036). Validated IHC antibody: PB9036 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Smooth muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissues; fixative not specified (datasheet PB9036); 2. Fixation: Image fixative and duration unreported (datasheet PB9036); 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 , 20 min (datasheet PB9036); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9036); 8. Primary antibody: Rabbit anti-LIF, 0.5-1μg/ml (datasheet PB9036); 9. Primary incubation: Overnight at 4 °C (datasheet PB9036). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet PB9036); verify before use.; Caveat: Secreted LIF may stain away from its producer cells (HPA tissue IHC; UniProt); Isoform / epitope: 2 isoforms; signal peptide 1–22 is removed; epitope unreported (UniProt); Regulation: Regulation not specified in the record (UniProt). Evidence: UniProt P15018, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the cytosol., Datasheet PB9036, PMC3655613, PMC12911375. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LIF/immunohistochemistry-ihc-guide.