ITLN1 / Intelectin-1 · IHC design guide

Design Immunohistochemistry for ITLN1

Plan chromogenic IHC on paraffin sections using intestinal goblet-cell cytoplasmic staining as a reference (HPA tissue IHC). The guide covers the catalog antibody’s 0.5–1 μg/mL IHC range (datasheet PB10004) and interpretation of staining from a secreted protein (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for ITLN1 (IHC for ITLN1): expected localisation Intestinal goblet-cell cytoplasm (HPA tissue IHC), antibody PB10004, validated IHC image, and IHC protocol steps — Printable **ITLN1** IHC protocol sheet — expected localisation Intestinal goblet-cell cytoplasm (HPA tissue IHC), antibody PB10004, controls and protocol steps. Open the full ITLN1 IHC guide →

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ITLN1 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	Intestinal goblet-cell cytoplasm (HPA tissue IHC)
	Staining pattern	Intestinal goblet cells show cytoplasmic staining (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet PB10004)
	Positive control ⓘ	Colon+4 more · see all
	Negative control ⓘ	Adipose tissue+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB10004)
	Caveat	Secreted protein may stain away from its RNA source (HPA tissue IHC)
	Regulation	High in intestine; low in subcutaneous fat (UniProt)
	Isoform / epitope	No annotated isoforms; mature chain is 19–298 (UniProt)

Section 1

Recommended ITLN1 IHC & IF Protocols

Compare the catalog antibody’s IHC protocol (datasheet PB10004) with a published rectal cancer biopsy protocol (PMC11740546).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet PB10004)
Fixation	Image fixative and duration unreported (datasheet PB10004); 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 PB10004); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet PB10004)
Primary antibody	Rabbit anti-ITLN1, 0.5-1μg/ml (datasheet PB10004)
Primary incubation	Overnight at 4 °C (datasheet PB10004)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB10004)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	ITLN1-positive staining in goblet cells of colon (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in intestinal goblet cells. No signal in the no-primary control.

💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet PB10004); the published study does not specify retrieval conditions (PMC11740546).

Section 2

What Is the Expected ITLN1 Staining Pattern?

ITLN1 is secreted and associated with the cell membrane despite having no transmembrane segment (UniProt Q8WWA0 topology). In paraffin section IHC, expect predominantly cytoplasmic staining in intestinal goblet cells (HPA: tissue IHC, Enhanced reliability). Interpret other staining cautiously: HPA reports medium consistency with RNA data, possible detection of protein from more than one gene, and presumed off target binding (HPA: reliability description).

What am I looking at on my slide?
Strong cytoplasmic staining in colon, duodenal, rectal, or small intestinal goblet cells.	This matches the principal tissue pattern; HPA scores goblet cells in each of these sites High (HPA: tissue IHC). Judge the cell type and compartment together rather than treating any intestinal color as a positive result.
Prominent nuclear staining, with little goblet cell cytoplasmic signal.	A nuclear dominant pattern does not match the reported goblet cell cytoplasmic pattern (HPA: tissue IHC). Treat it as suspect staining; inspect the negative control and repeat with optimized detection before assigning it to ITLN1.
Strong staining in adipocytes or unrelated cells while nearby goblet cells are weak.	HPA reports adipocytes as Not detected, while UniProt places omental ITLN1 in stromal vascular cells rather than fat cells (HPA: tissue IHC; UniProt Q8WWA0 tissue specificity). Consider cross reactivity or endogenous detection activity and compare appropriate controls.
Widespread color obscures cell boundaries and goblet cell contrast.	A diffuse field cannot establish the cell restricted pattern reported by HPA (HPA: tissue IHC). Compare a reagent negative control, then review blocking, antibody concentration, washing, and chromogen development as general IHC troubleshooting steps.
No signal in a section containing identifiable intestinal goblet cells.	That conflicts with the High goblet cell staining reported for colon, duodenum, rectum, and small intestine (HPA: tissue IHC). Confirm tissue preservation and run a known positive section in the same batch before interpreting the sample as negative.

💡Expected ITLN1 appearanceCall the IHC result positive when goblet cell cytoplasm is distinctly High against surrounding tissue (HPA: tissue IHC); diffuse, nuclear dominant, or adipocyte dominant color is suspect (HPA: tissue IHC).

How each factor affects the staining
Tissue and cell selection	Colon, duodenum, rectum, and small intestine offer High goblet cell staining; appendix goblet cells are Medium (HPA: tissue IHC). Adipocytes are Not detected (HPA: tissue IHC). Score the named cells, not a whole section average.
Secretion and topology	ITLN1 is secreted and cell membrane associated, with no transmembrane segment (UniProt Q8WWA0 topology). HPA notes that a secreted protein's RNA and protein tissue locations can differ (HPA: reliability description); use observed IHC localization to interpret a slide.
Evidence and antibody choice	The tissue profile has Enhanced reliability, but HPA warns of medium RNA–staining consistency, possible detection of protein from more than one gene, and disregarded presumed off target binding (HPA: tissue IHC). HPA063275 is IHC Enhanced; HPA067326 and CAB012652 are IHC Supported (HPA: antibodies).
Processing and epitope interpretation	UniProt lists a signal peptide at residues 1–18, a chain at 19–298, a propeptide at 299–313, and one glycosylation site at residue 163 (UniProt Q8WWA0 processing). Without an antibody epitope map, these features do not predict a specific IHC staining difference.
What should IF/ICC show?	HPA reports vesicles and the sperm equatorial segment as main approved locations, with acrosome, mid piece, and principal piece as additional approved locations (HPA: subcellular ICC-IF). Those observations are a separate IF/ICC reference, not an alternative paraffin section IHC pattern.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Goblet cells are unstained in the positive control.	The run may have a detection, retrieval, or antibody setup problem; HPA reports High intestinal goblet cell staining (HPA: tissue IHC).	Check the positive control, antibody identity, detection reagents, and retrieval conditions together; adjust one general IHC workflow variable at a time. No ITLN1 specific retrieval requirement is supplied.
The entire section has diffuse chromogen.	Excess reagent signal or incomplete washing can obscure cell localization in standard IHC; the expected pattern is goblet cell cytoplasmic (HPA: tissue IHC).	Compare a reagent negative control, review blocking and wash steps, and reduce antibody or chromogen exposure if the controls indicate background.
Adipocytes show strong staining.	This disagrees with Not detected adipocytes in HPA IHC and with UniProt's omental stromal vascular cell localization (HPA: tissue IHC; UniProt Q8WWA0 tissue specificity).	Check cell identity on the counterstain and compare negative controls; investigate cross reactivity or endogenous detection activity before scoring adipocytes as ITLN1 positive.
Nuclei dominate the signal.	The observed compartment differs from HPA's cytoplasmic intestinal goblet cell profile (HPA: tissue IHC).	Review morphology and negative controls, then repeat the stain with controlled detection conditions; require a reproducible goblet cell cytoplasmic pattern for the intended IHC call.
A nonintestinal section stains, but the intestinal control does not.	A positive color alone is weak evidence when the known goblet cell pattern fails; HPA also cautions about possible detection of protein from more than one gene (HPA: tissue IHC, reliability description).	Resolve the failed positive control first. Then compare tissue morphology and an independent IHC validated antibody where available (HPA: antibodies).
Protein staining seems inconsistent with tissue RNA.	HPA reports medium RNA–staining consistency and cautions that secreted protein and RNA can occupy different tissue locations (HPA: reliability description); UniProt lists ITLN1 as secreted (UniProt Q8WWA0 subcellular location).	Score the protein signal by cell type and compartment, document the discrepancy, and use controls or an independent antibody to assess staining specificity (HPA: antibodies).

Source: UniProt Q8WWA0 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing ITLN1 staining

IHC analysis of ITLN1 using anti-ITLN1 antibody (PB10004). ITLN1 was detected in a paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-ITLN1 Antibody (PB10004) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

PB10004 · IHCIHC analysis of ITLN1 using anti-ITLN1 antibody (PB10004). ITLN1 was detected in a paraffin-embedded section of human intestinal cancer tiss

IF analysis of ITLN1 using anti-ITLN1 antibody (PB10004). ITLN1 was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-ITLN1 Antibody (PB10004) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

PB10004 · IFIF analysis of ITLN1 using anti-ITLN1 antibody (PB10004). ITLN1 was detected in immunocytochemical section of U20S cells. Enzyme antigen ret

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000179914-ITLN1/tissue/cerebral+cortex#imid_17639683

HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000179914-ITLN1/tissue/duodenum#imid_17639608

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000179914-ITLN1/tissue/colon#imid_17639601

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000179914-ITLN1/tissue/liver#imid_17639602

HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA063275; green = ITLN1, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for ITLN1 IHC & IF

🧪Run colon first: goblet cells should stain strongly (HPA: High in colon goblet cells). Use adrenal gland as a negative tissue (HPA: Not detected in adrenal gland glandular cells); compare neighboring nongoblet cells on the colon slide with the no-primary background rather than assuming they are ITLN1-negative.

Positive control tissue: Colon (Goblet cells, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ITLN1 in HEL, Sperm, with annotated localisation: Vesicles (approved), Equatorial segment (approved) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) slide, a rabbit IgG isotype control matched to the primary antibody’s format (caption: rabbit anti-ITLN1), and an ITLN1-knockout biological negative if available. For colon, check endogenous peroxidase and biotin background before interpreting the biotin-based DAB signal (caption: biotinylated secondary and SABC–DAB).

⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the fixative for the PB10004 paraffin-section image is unreported (caption: fixative not stated). The caption uses heat-mediated EDTA retrieval at pH 8.0, but does not establish that retrieval is required (caption: EDTA retrieval). Whether frozen sections or IF/ICC are easier is unreported; assess staining around mucus-rich colon goblet cells against the background controls (HPA: High in colon goblet cells).

HPA tissue IHC evidence for ITLN1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. Secreted protein, tissue location of RNA and protein is expected to differ. Presumed off target binding observed and disregarded.). 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
Colon	Goblet cells	High	Protein (IHC)	HPA →
Duodenum	Goblet cells	High	Protein (IHC)	HPA →
Rectum	Goblet cells	High	Protein (IHC)	HPA →
Small intestine	Goblet cells	High	Protein (IHC)	HPA →
Testis	Spermatogonia 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 →
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced ITLN1 IHC Tips

Troubleshoot ITLN1 staining in paraffin sections by checking retrieval, cell identity, and compartment before comparing staining intensity across samples.

What retrieval should I try first if ITLN1 staining is weak?

Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet PB10004). The selected paraffin-section image used this retrieval before staining with 1 μg/ml catalog antibody overnight at 4°C (caption PB10004). If signal is weak, keep the antibody concentration and detection method constant while adjusting heating time and checking whether sections remain attached (standard IHC practice). Include a section containing intestinal goblet cells as a positive reference, since these cells show high ITLN1 staining (HPA: colon, duodenum, rectum and small intestine goblet cells). Document any alternative retrieval buffer as a fallback tested on matched sections, because altered retrieval can change both signal and background (standard IHC practice).

Could fixation explain inconsistent ITLN1 staining between paraffin blocks?

Target-specific sensitivity of ITLN1 to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (caption PB10004). Record the fixative, fixation duration, tissue thickness and processing history for each block before attributing differences to biology (standard IHC practice). Compare matched sections using EDTA at pH 8.0 and the same antibody and detection conditions, so processing is the main variable under review (datasheet PB10004; standard IHC practice). Use an intestinal goblet-cell reference in each run to detect changes in assay performance (HPA: high goblet-cell staining; standard IHC practice). Avoid treating a weak block alone as evidence of reduced ITLN1 expression (standard IHC practice).

Which staining compartments are plausible for ITLN1 in tissue sections?

Expect cytoplasmic staining in intestinal goblet cells when assessing tissue IHC, since that is the reported tissue pattern (HPA: cytoplasmic expression in intestinal goblet cells). ITLN1 is annotated as secreted and cell-membrane associated, enriched in lipid rafts, with no transmembrane segment (UniProt Q8WWA0: subcellular location and topology). Thus, record intracellular, apical and extracellular staining separately instead of calling every stained region a positive cell (UniProt Q8WWA0: location; standard IHC practice). Check whether apparent membrane outlines follow intact cells or deposited material, using the counterstain and adjacent sections (standard IHC practice). The reported vesicular and sperm-region IF locations should not be imposed on every paraffin tissue compartment (HPA: subcellular IF locations).

How should epitope uncertainty affect ITLN1 IHC troubleshooting?

Do not assume an isoform-specific staining pattern: the supplied record lists 0 isoforms and gives no antibody epitope (UniProt Q8WWA0: isoforms; caption PB10004). The annotated mature chain spans residues 19–298, with a signal peptide at 1–18 and a propeptide at 299–313 (UniProt Q8WWA0: processing). One glycosylation site is annotated at residue 163, but its effect on this antibody's tissue staining has not been established (UniProt Q8WWA0: glycosylation; caption PB10004). If staining changes with retrieval, compare matched sections and examine cell-level localisation before proposing epitope masking (standard IHC practice). Seek epitope documentation or an independent antibody before assigning a discrepant pattern to protein processing (standard IHC practice).

How can I check the IHC localisation with a separate IF experiment?

For a separate IF experiment, pair ITLN1 with a validated goblet-cell marker when examining intestine, where goblet cells show high tissue staining (HPA: intestine goblet cells; standard IF practice). Choose a fluorophore channel with low tissue autofluorescence and include unstained and secondary-only controls before interpreting dim signal (standard IF practice). ITLN1 is secreted or membrane associated and lacks a transmembrane segment, while IF annotations also report vesicles; the antibody's epitope side is unspecified (UniProt Q8WWA0: location and topology; HPA: subcellular location; caption PB10004). Compare permeabilised and non-permeabilised preparations if intracellular versus accessible surface signal matters (standard IF practice). Treat the paraffin IHC retrieval and chromogenic conditions as evidence for this IHC assay only (caption PB10004).

How do I separate ITLN1 staining from chromogenic background?

Inspect no-primary and secondary-only sections alongside the ITLN1 stain to identify detection-system background (standard IHC practice). The selected image used a biotinylated secondary, streptavidin–biotin complex and DAB, so test the detection system's background on the same tissue type (caption PB10004; standard IHC practice). Apply a peroxidase block as a general chromogenic IHC step, and check whether pigment or precipitate persists without primary antibody (standard IHC practice). Compare background with the expected goblet-cell pattern rather than accepting diffuse colour as ITLN1 signal (HPA: cytoplasmic goblet-cell expression; standard IHC practice). If background rises with stronger retrieval or antibody concentration, adjust one condition at a time on matched sections (standard IHC practice).

What should I measure when comparing ITLN1 across sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the scored compartment before analysis: intestinal goblet cells are a documented positive population, whereas adipocytes are reported as undetected (HPA: goblet-cell and adipocyte tissue IHC). For goblet-cell staining, report the percentage of positive goblet cells and an H-score using intensity grades 0–3, giving a possible range of 0–300 (standard IHC scoring practice). Normalise cell-based scores to the number of evaluable goblet cells, and report stained-area density per mm² only for a consistently defined tissue region (standard IHC practice). Keep retrieval, detection, counterstain and imaging settings consistent across compared sections (standard IHC practice). Exclude folds, necrosis and damaged edges using rules fixed before scoring (standard IHC practice).

When is an unexpected ITLN1-positive region likely to be artefact?

A convincing intestinal signal should track identifiable goblet cells and their cytoplasm, the reported tissue IHC pattern (HPA: cytoplasmic intestinal goblet-cell expression). Interpret staining in adipocytes cautiously because they are reported as undetected, while omental ITLN1 is described in stromal vascular cells rather than fat cells (HPA: adipocytes not detected; UniProt Q8WWA0: tissue specificity). Check edge-only colour, necrotic regions and DAB deposits against intact central tissue and no-primary controls (standard IHC practice). Signal in an unexpected cell or compartment warrants confirmation with an independent antibody or another assay before assigning it to ITLN1 (standard IHC practice). HPA also notes presumed off-target binding and that secreted protein can appear away from its RNA-producing cells (HPA: tissue-IHC reliability description).

Boster reagents

Best ITLN1 / Intelectin-1 IHC Antibodies

PB10004 has human-reactive IHC and IF/ICC applications (catalog: PB10004); its figures show paraffin-section IHC in human intestinal cancer tissue and IF/ICC in U20S cells (PB10004 image captions).

Real IHC data

Anti-ITLN1 Antibody ®

Cat # PB10004

View product →

PB10004 is the only SKU shown (catalog: PB10004); its IHC figure uses a paraffin-embedded section of human intestinal cancer tissue (PB10004 IHC caption). Its IF/ICC figure uses U20S cells (PB10004 IF caption).

Which to pick: Choose PB10004 for tissue IHC: its own figure documents paraffin-section staining, but does not report the fixative (PB10004 IHC caption). Choose PB10004 for IF/ICC based on its listed applications and U20S-cell figure (catalog: PB10004; PB10004 IF caption). No cross-species option is documented: PB10004 lists human reactivity only, and its clonality is unspecified (catalog: PB10004).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry Q8WWA0 (ITLN1_HUMAN, Intelectin-1).
Human Protein Atlas. ITLN1 tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. ITLN1 subcellular location (ICC-IF): Mainly localized to vesicles and equatorial segment. In addition localized to the acrosome, mid piece and principal piece..
Human Protein Atlas. ITLN1 antibody validation summary (3 antibodies).
High Intelectin-1 Expression Associated with Aggressive Tumor Behavior and Worse Survival in Rectal Cancer. OncoTargets and therapy 2025 — PMC11740546.
Characterization of an intelectin-1 (Itln1) knockout mouse model. Frontiers in immunology 2022 — PMC9441953.
Value analysis of ITLN1 in the diagnostic and prognostic assessment of colorectal cancer. Translational cancer research 2024 — PMC11231763.
Omentin-1 promotes wound healing in diabetic mice by improving vascular endothelial cell function. Scientific reports 2025 — PMC12635396.
PubMed PMID:11747454 — UniProt-cited evidence.
PubMed PMID:11181563 — UniProt-cited evidence.
PubMed PMID:11313366 — UniProt-cited evidence.