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LITAF / Lipopolysaccharide-induced tumor necrosis factor-alpha factor · IHC design guide
Design Immunohistochemistry for LITAF
Plan chromogenic IHC for LITAF in paraffin sections using its cytoplasmic tissue pattern and strong lymphoid staining (HPA tissue IHC). This guide covers positive controls, antibody dilution, and interpretation of macrophage-rich regions (HPA tissue IHC; datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LITAF 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; endosomal membranes are expected (HPA tissue IHC; UniProt) | |
| Staining pattern | Cytoplasmic staining, most abundant in lymphoid tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Esophagus+3 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 | Macrophage-rich regions may dominate the signal (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | 3 isoforms; extracellular versus cytoplasmic epitope topology is unassigned (UniProt) |
Section 1
Recommended LITAF IHC & IF Protocols
The catalog antibody IHC-P protocol is accompanied by four published LITAF IHC procedures covering pancreatic specimens, tonsil follicles, and mouse brain (PMC5790452; PMC4111142; PMC10447466; PMC11705406).
| Sample | Paraffin-embedded human prostate tissue; fixative not specified (datasheet A03043) |
| Fixation | Image fixative and duration unreported (datasheet A03043); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-LITAF, 1:50-1:200 (datasheet A03043) |
| 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 | LITAF-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, most abundant in lymphoid tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval); use citrate when reproducing a cited procedure (PMC5790452; PMC11705406).
Section 2
What Is the Expected LITAF Staining Pattern?
LITAF is membrane-associated in endosomal and lysosomal compartments and can enter the nucleus despite having no transmembrane segment (UniProt Q99732 localization and topology). In tissue IHC, expect mainly cytoplasmic staining, especially in lymphoid tissue, lung macrophages, and bone marrow hematopoietic cells (HPA tissue IHC). HPA rates its tissue IHC evidence Approved, with medium consistency between staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining is strong in lung macrophages, bone marrow hematopoietic cells, or lymph node non-germinal center cells (HPA tissue IHC: High). | This fits the reported tissue pattern (HPA tissue IHC). Compare cell populations within the section: the expected result is cellular staining with recognizable boundaries, while reported intensity varies by tissue and cell type (HPA tissue IHC; general IHC practice). |
| Punctate or perinuclear cytoplasmic staining accompanies a broader cytoplasmic signal (UniProt Q99732 localization; HPA tissue IHC). | An endosomal or lysosomal distribution is plausible because LITAF associates with those membranes (UniProt Q99732 localization). Tissue IHC alone cannot identify the stained organelle; reserve that assignment for localization evidence beyond chromogenic morphology (general IHC practice). |
| Strong staining appears only in an unexpected compartment, such as extracellular material (UniProt Q99732 localization). | Review it as possible nonspecific signal because extracellular localization is absent from the supplied record (UniProt Q99732 localization). Nuclear staining alone is inconclusive: UniProt reports nuclear translocation, and HPA ICC-IF supports nucleoplasmic localization (UniProt Q99732; HPA subcellular ICC-IF). |
| Squamous epithelium or placental trophoblasts stain strongly while expected positive cells remain faint (HPA tissue IHC: Not detected in those cell types). | The distribution conflicts with HPA tissue IHC and warrants checks for antibody cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). Interpret placenta by cell type: UniProt reports placental expression, whereas HPA reports trophoblasts as undetected (UniProt Q99732 tissue specificity; HPA tissue IHC). |
| Color spreads across many cell types or obscures tissue architecture, including areas expected to be weak (HPA tissue IHC: variable cell-type levels). | Diffuse background limits compartment and cell-type scoring (general IHC practice). Compare the test section with an appropriate detection control before attributing widespread color to LITAF; low or absent HPA staining is a useful comparison, not proof of a universally negative tissue (HPA tissue IHC; general IHC practice). |
💡Expected LITAF appearanceCall a section positive when identifiable cells show predominantly cytoplasmic staining, strongest in reported high-expression populations; broad background or strong signal confined to HPA-undetected cell types needs review (HPA tissue IHC; general IHC practice).
| Compartment and topology (UniProt Q99732; HPA subcellular ICC-IF) | LITAF associates with endosome and lysosome membranes but has no transmembrane segment (UniProt Q99732). HPA tissue IHC describes general cytoplasmic expression; HPA ICC-IF additionally supports nucleoplasm, nuclear speckles, Golgi apparatus, and plasma membrane localization (HPA tissue IHC; HPA subcellular ICC-IF). |
| Cell-type variation (HPA tissue IHC) | HPA reports High staining in lung macrophages, spleen red-pulp cells, and tonsil non-germinal center cells, but Medium staining in adipocytes and Low staining in several neuronal or glandular populations (HPA tissue IHC). Score the relevant cells rather than averaging an entire mixed tissue (general IHC practice). |
| Placental evidence (UniProt Q99732; HPA tissue IHC) | UniProt describes placenta among predominant expression sites, while HPA reports trophoblastic cells as Not detected by tissue IHC (UniProt Q99732 tissue specificity; HPA tissue IHC). Neither statement identifies every placental cell population; avoid treating trophoblasts as an assured positive control (HPA tissue IHC). |
| Isoforms and processing (UniProt Q99732) | UniProt lists 3 isoforms and a chain spanning residues 1–161, with no signal peptide or propeptide (UniProt Q99732). The supplied evidence gives no antibody epitope or isoform coverage, so a staining difference cannot be assigned to a particular isoform (UniProt Q99732; supplied antibody record). |
| Antibody validation (HPA antibodies; HPA tissue IHC) | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| IF/ICC Q: Should the same compartment rule apply? (HPA subcellular ICC-IF) | A: Evaluate IF/ICC against its own localization evidence: HPA supports nucleoplasm and also reports nuclear speckles, Golgi apparatus, vesicles, and plasma membrane (HPA subcellular ICC-IF). This tissue IHC pattern is a separate readout (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in lung macrophages or bone marrow hematopoietic cells (HPA tissue IHC: High). | The result conflicts with reported high staining; the cause is undetermined from the tissue profile (HPA tissue IHC). | Check section integrity, antibody application, retrieval, and detection controls using the catalog antibody’s IHC-P instructions where available (general IHC practice); do not infer LITAF-specific retrieval sensitivity (HPA tissue IHC). |
| Strong color occurs throughout a section, including HPA-undetected squamous epithelium (HPA tissue IHC). | Nonspecific binding or endogenous detection activity could contribute (general IHC practice). | Review a no-primary control and the detection system’s appropriate blocking step; compare staining by cell type with the HPA profile (general IHC practice; HPA tissue IHC). |
| Nuclear staining accompanies cytoplasmic staining (UniProt Q99732; HPA tissue IHC). | Nuclear localization is possible, although tissue IHC chiefly reports cytoplasmic expression (UniProt Q99732; HPA tissue IHC). | Record nuclear and cytoplasmic scores separately and inspect morphology and controls before interpreting the nuclear component (general IHC practice); HPA ICC-IF supports nucleoplasmic localization (HPA subcellular ICC-IF). |
| Trophoblasts are negative in an otherwise stained placental section (HPA tissue IHC: Not detected). | That cell-type result matches HPA, even though UniProt lists placenta among predominant expression sites (HPA tissue IHC; UniProt Q99732 tissue specificity). | Identify the cells being scored and choose a reported high-staining population for a positive comparison (HPA tissue IHC; general IHC practice). |
| Signal is patchy across lymphoid or mixed-cell tissue (HPA tissue IHC: cell-type variation). | Different populations can have different reported levels; HPA highlights non-germinal center cells in lymph node and tonsil (HPA tissue IHC). | Score named cell populations separately and compare like regions across sections (HPA tissue IHC; general IHC practice). |
| Fine vesicle-like staining cannot be assigned confidently on chromogenic IHC (UniProt Q99732 localization). | LITAF has several reported membrane-associated locations, and chromogenic morphology alone may not resolve them (UniProt Q99732; general IHC practice). | Report the visible cytoplasmic pattern without naming an organelle; use the separate IF/ICC guide when subcellular localization is the question (HPA subcellular ICC-IF; general IHC practice). |
Experiment evidence
IHC & IF images showing LITAF staining
A03043 · IHCImmunohistochemistry of paraffin-embedded human prostate using LITAF Antibody at dilution of 1:100 .Perform microwave antigen retrieval with
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000189067-LITAF/tissue/cerebral+cortex#imid_2165660
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000189067-LITAF/tissue/colon#imid_2165578
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000189067-LITAF/tissue/liver#imid_2165579
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000189067-LITAF/tissue/kidney#imid_2165580
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA006960; green = LITAF, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA006960; green = LITAF, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LITAF IHC & IF
🧪Run prostate first: its glandular cells must stain, with a matched paraffin-section example available (HPA: High in prostate glandular cells; SKU A03043 caption). Use esophageal squamous epithelium as the negative tissue (HPA: Not detected in esophageal squamous epithelial cells); on the prostate slide, treat cells without staining as internal background comparators only if present, since HPA identifies no internal negative cell population there (HPA: prostate glandular cells High).
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Esophagus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LITAF in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and an isotype control matched to the primary antibody’s host species and clonality; use LITAF-knockout material or an immunogen-peptide block, if available, to test target specificity (standard IHC practice). For chromogenic detection, block endogenous peroxidase and check the prostate no-primary slide for background signal (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected paraffin-section caption does not state a fixative (SKU A03043 caption). That example uses microwave retrieval in 10 mM PBS, pH 7.2, before staining at 1:100; it supports a starting condition but does not establish retrieval dependence (SKU A03043 caption). Whether frozen sections or IF are easier is unreported; inspect prostate gland lumina for nonspecific chromogen deposits when scoring glandular cells (standard IHC practice; HPA: High in prostate glandular cells).
HPA tissue IHC evidence for LITAF
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Prostate | Glandular cells | High | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LITAF IHC Tips
Troubleshoot LITAF staining by comparing compartment, cell type, and control sections with the documented paraffin IHC image and tissue expression data.
Which retrieval condition should I try first for LITAF paraffin IHC?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule). If staining is weak or tissue morphology suffers, compare serial sections using microwave retrieval in 10 mM PBS, pH 7.2, the condition documented for the catalog antibody in paraffin-embedded prostate at 1:100 (A03043 caption). Keep section thickness, detection chemistry, and development time matched so retrieval is the principal variable (standard IHC practice). Judge the result in glandular cells and against a no-primary control, while checking that stronger signal does not simply reflect increased background (HPA: High in prostate glandular cells; standard IHC practice).
How should I troubleshoot fixation-related loss of LITAF staining?
The catalog image documents paraffin-embedded prostate but does not report its fixative, so target-specific LITAF fixation sensitivity is unknown (A03043 caption). Record fixative, fixation duration, processing history, and section age for each specimen before comparing staining outcomes (standard IHC practice). On matched sections, first hold retrieval and the 1:100 antibody dilution constant, then assess whether altered processing correlates with reduced signal or damaged morphology (A03043 caption; standard IHC practice). Include a comparably processed positive tissue and a no-primary section; a failed positive control points to a workflow problem but does not establish a LITAF-specific fixation effect (HPA: High in prostate glandular cells; standard IHC practice).
Should LITAF stain the cytoplasm, nucleus, or both?
Evaluate cytoplasmic staining first in tissue sections because the tissue IHC profile reports general cytoplasmic expression, most abundant in lymphoid tissues (HPA tissue IHC). Nuclear staining remains plausible: LITAF can move from cytoplasm to nucleus, and cell imaging reports nucleoplasm as its main location (UniProt Q99732 subcellular; HPA subcellular). Vesicular, Golgi, and membrane-associated patterns also fit reported locations, although chromogenic IHC may not resolve individual small vesicles (HPA subcellular; standard IHC practice). Compare the same cell population across serial sections and controls; a sharp nuclear-only or diffuse extracellular pattern needs independent antibody or orthogonal evidence before assignment to LITAF (standard IHC practice).
Can this stain distinguish LITAF isoforms or reveal a masked epitope?
The record lists 3 LITAF isoforms, a 161-residue reference chain, and a LITAF domain spanning residues 76–160 (UniProt Q99732 record). No antibody epitope or isoform-specific binding information is supplied, so chromogenic staining cannot be assigned to one isoform from this evidence (A03043 caption; UniProt Q99732 record). The record lists no signal peptide, propeptide, transmembrane segment, or glycosylation site; none of those annotations establishes whether fixation masks this antibody’s epitope (UniProt Q99732 record). If retrieval changes the pattern, compare matched sections and seek an independently mapped antibody or isoform-resolving assay before interpreting the difference as isoform expression (standard IHC practice).
How can I check LITAF localisation with multiplex IF?
For a secondary IF experiment, pair LITAF with a validated marker for the cell population under study, such as a macrophage marker when examining lung macrophages (HPA: High in lung macrophages; standard IF practice). Choose spectrally separated fluorophores and place the weaker signal in a far-red channel when tissue autofluorescence interferes, then inspect single-stain and no-primary controls (standard IF practice). Because the antibody epitope and its membrane-facing side are unspecified, compare mild permeabilisation with an unpermeabilised condition before treating a missing vesicular or nuclear signal as biological absence (A03043 caption; UniProt Q99732 topology; standard IF practice). Keep IF conclusions separate from the paraffin IHC image, which documents chromogenic tissue staining only (A03043 caption).
What explains diffuse or patchy background in LITAF chromogenic IHC?
First compare a no-primary section with the stained section to identify secondary-reagent signal, endogenous enzyme activity, or pigment that persists without primary antibody (standard IHC practice). For peroxidase-based chromogenic detection, check the peroxide block and shorten DAB development if background rises across many cell types; these are general workflow adjustments (standard IHC practice). Titrate the catalog antibody around its documented 1:100 prostate-image dilution while keeping retrieval and detection constant, and assess tissue edges separately from intact interiors (A03043 caption; standard IHC practice). Broad cytoplasmic staining alone is inconclusive because LITAF is widely expressed; use cell-level patterns and controls to distinguish plausible signal from nonspecific deposit (UniProt Q99732 tissue specificity; HPA tissue IHC; standard IHC practice).
How should I quantify heterogeneous LITAF staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring, since tissue IHC describes predominantly cytoplasmic expression while cell imaging also supports nucleoplasmic localisation (HPA tissue IHC; HPA subcellular). For chromogenic sections, report percent positive cells and an H-score from 0–300, calculated from the percentages at intensity grades 0–3 (standard IHC practice). If counting discrete positive cells, report density per mm² of evaluable tissue and exclude necrotic or folded areas using the same rule for every section (standard IHC practice). Normalise cell-based measures to the relevant identified cell population, and compare batches only after checking matched controls and comparable DAB development (standard IHC practice).
When is an unexpected LITAF-positive area likely to be artefact?
Check whether staining lies inside intact cells and matches a plausible compartment: cytoplasm in tissue, with nuclear or vesicular localisation also supported by other records (HPA tissue IHC; HPA subcellular). Strong staining in lung macrophages or prostate glandular cells has cell-level support, whereas signal confined to squamous epithelial cells of esophagus or oral mucosa warrants extra scrutiny (HPA: High in lung macrophages and prostate glandular cells; HPA: Not detected in esophagus and oral mucosa squamous epithelial cells). Inspect edges, necrotic regions, and the no-primary control for staining caused by section damage or endogenous enzyme activity (standard IHC practice). Treat a positive call as provisional when compartment, cell identity, and controls disagree; verify it with independent evidence (standard IHC practice).
Boster reagents
Best LITAF / Lipopolysaccharide-induced tumor necrosis factor-alpha factor IHC Antibodies
The catalog lists human- and mouse-reactive LITAF antibodies for IHC (catalog: A03043, A03043-2), with a paraffin-section image for A03043 (A03043 image caption). A03043-2 also lists rat reactivity and IF/ICC (catalog: A03043-2).
Real IHC data
Anti-LITAF Antibody
Cat # A03043
A03043 lists human and mouse IHC reactivity and has a human prostate paraffin-section IHC image at 1:100 (catalog: A03043; A03043 image caption). A03043-2 lists human, mouse and rat IHC reactivity plus human IF/ICC, but has no IHC or IF image in the supplied catalog (catalog: A03043-2).
Which to pick: For tissue IHC, choose A03043 when a matching paraffin-section example matters: its human prostate caption specifies microwave retrieval in 10 mM PBS at pH 7.2, but does not report the fixative (A03043 image caption). For IF/ICC, choose A03043-2, which lists human IF/ICC at 5 μg/ml without an IF image (catalog: A03043-2). For rat tissue IHC, A03043-2 lists rat reactivity and IHC at 2–5 μg/ml; both catalog antibodies are rabbit polyclonals (catalog: A03043, A03043-2).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q99732 (LITAF_HUMAN, Lipopolysaccharide-induced tumor necrosis factor-alpha factor).
- Human Protein Atlas. LITAF tissue IHC expression (reliability: Approved).
- Human Protein Atlas. LITAF subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nuclear speckles, plasma membrane, the Golgi apparatus and vesicles..
- Human Protein Atlas. LITAF antibody validation summary (1 antibodies).
- LITAF is a potential tumor suppressor in pancreatic cancer. Oncotarget 2018 — PMC5790452.
- LITAF, a BCL6 target gene, regulates autophagy in mature B-cell lymphomas. British journal of haematology 2013 — PMC4111142.
- Single cell transcriptomic analyses implicate an immunosuppressive tumor microenvironment in pancreatic cancer liver metastasis. Nature communications 2023 — PMC10447466.
- Impact of LITAF on Mitophagy and Neuronal Damage in Epilepsy via MCL-1 Ubiquitination. CNS neuroscience & therapeutics 2025 — PMC11705406.
- PubMed PMID:9305847 — UniProt-cited evidence.
- PubMed PMID:10200294 — UniProt-cited evidence.
- PubMed PMID:11274176 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LITAF (Lipopolysaccharide-induced tumor necrosis factor-alpha factor). Expected localisation Cytoplasmic tissue staining; endosomal membranes are expected (HPA tissue IHC; UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A03043 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Esophagus. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate tissue; fixative not specified (datasheet A03043); 2. Fixation: Image fixative and duration unreported (datasheet A03043); 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 Tris-EDTA buffer, pH 9.0 , 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-LITAF, 1:50-1:200 (datasheet A03043); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Macrophage-rich regions may dominate the signal (HPA tissue IHC); Isoform / epitope: 3 isoforms; extracellular versus cytoplasmic epitope topology is unassigned (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt Q99732, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A03043, PMC5790452, PMC4111142. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LITAF/immunohistochemistry-ihc-guide.