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JAG1 / Protein jagged-1 · IHC design guide
Design Immunohistochemistry for JAG1
Plan chromogenic JAG1 IHC on paraffin sections around its observed cytoplasmic and membranous pattern (HPA tissue IHC). Start the IHC-validated antibody at 5 μg/mL (datasheet: A00640), using gallbladder glandular cells as a high-staining reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
JAG1 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 and membranous in tissue (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic and membranous staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Gallbladder+4 more · see all | |
| Negative control | Bone marrow+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has medium consistency with RNA (HPA tissue IHC) | |
| Regulation | Higher in cervical squamous carcinoma (UniProt) | |
| Isoform / epitope | 2 isoforms; map extracellular vs cytoplasmic epitopes (UniProt) |
Section 1
Recommended JAG1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) is followed by published JAG1 staining protocols for colorectal tumors (PMC5132269), liver lesions (PMC5177771), and frozen patient tissue (PMC4988718).
| Sample | Paraffin-embedded Rat liver tissue; fixative not specified (datasheet M00640-1) |
| Fixation | Image fixative and duration unreported (datasheet M00640-1); 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 citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane 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 monoclonal (clone 17J22) anti-JAG1, 1:50-1:200 (datasheet M00640-1) |
| 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 | JAG1-positive staining in glandular cells of gallbladder (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic and membranous expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen retrieval); the paraffin-section studies used citrate with autoclave or microwave heating (PMC5132269; PMC5177771).
Section 2
What Is the Expected JAG1 Staining Pattern?
JAG1 is a cell-membrane protein with an extracellular region and a cytoplasmic tail (UniProt P78504 topology). In paraffin-section IHC, expect cytoplasmic and membranous staining, especially in gallbladder glandular cells and skin eccrine glands (HPA tissue IHC: High). HPA rates the tissue pattern Supported, with medium consistency between antibody staining and RNA expression; assess the pattern by cell type as well as compartment (HPA tissue IHC).
| Membranous and cytoplasmic staining in gallbladder glandular cells or skin eccrine glands. | This fits the reported high-staining cell populations and HPA's overall cytoplasmic and membranous pattern (HPA tissue IHC: High; HPA tissue IHC profile). Compare signal with neighboring cells rather than expecting every cell in the section to stain equally (HPA tissue IHC: cell-specific levels). |
| Predominantly nuclear staining, with little signal in the expected compartments. | A nuclear-only pattern does not match JAG1's membrane topology or HPA's tissue IHC profile (UniProt P78504 topology; HPA tissue IHC profile). Treat it as questionable staining and check controls and detection conditions before scoring it as JAG1. |
| Strong staining in bone-marrow hematopoietic cells or caudate or hippocampal glial cells. | Those specified cell populations were reported as not detected (HPA tissue IHC: Not detected). Consider cross-reactivity or endogenous detection activity, then check negative controls. The HPA result applies to those cells, not every cell in each tissue. |
| Diffuse chromogen across cells and surrounding section, without a discernible cell pattern. | This is hard to reconcile with HPA's cell-resolved staining levels (HPA tissue IHC). Review background on the negative control and the blocking, washing, and detection steps (general IHC practice) before assigning a JAG1 score. |
| No detectable signal in gallbladder glandular cells or skin eccrine glands. | These are reported high-staining reference populations (HPA tissue IHC: High). Check whether tissue and run controls worked, then review the IHC-validated antibody's stated conditions (general IHC practice). Absence in one run does not establish biological absence. |
💡Expected JAG1 appearanceCall a positive result when identifiable gallbladder glandular cells or skin eccrine glands show high cytoplasmic and membranous staining (HPA tissue IHC: High; HPA tissue IHC profile); predominantly nuclear staining or strong signal in reported not-detected cell populations is suspect (UniProt P78504 topology; HPA tissue IHC: Not detected).
| Cell population and reference intensity | High staining is reported in gallbladder glandular cells and skin eccrine glands; medium staining occurs in several other listed glandular populations and colon endothelial cells (HPA tissue IHC). Choose a reference cell population, not merely a positive organ. |
| Topology and antibody epitope | JAG1 spans the membrane at residues 1068–1093, with an extracellular region at 34–1067 and a cytoplasmic region at 1094–1218 (UniProt P78504 topology). The antibody epitope is unspecified here, so the record does not support an epitope-specific staining prediction. |
| Isoforms and processing | UniProt lists 2 isoforms and a signal peptide at residues 1–33 (UniProt P78504). Without an antibody epitope or isoform-coverage record, do not attribute a staining difference to an isoform or processing event. |
| Strength of tissue IHC evidence | HPA calls its tissue IHC profile Supported and reports medium consistency with RNA expression; the listed IHC antibodies are Supported, not Enhanced (HPA tissue IHC; HPA antibodies). Treat a matching pattern as supporting evidence, not proof of specificity. |
| IF/ICC Q&A: Where should JAG1 appear? | HPA reports approved plasma-membrane and Golgi-apparatus localization in ICC-IF (HPA subcellular ICC-IF). This answers the localization question; IF/ICC conditions belong in its separate guide. Golgi staining alone does not redefine the paraffin-section IHC profile (HPA tissue IHC profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported high-staining reference cell population is blank. | The staining run or detection may have failed; one blank slide cannot establish JAG1 absence (HPA tissue IHC: High; general IHC practice). | Check tissue identity and run controls, then review the IHC-validated antibody's stated dilution, retrieval, and detection conditions (general IHC practice). No target-specific fixation sensitivity is reported here. |
| Staining is mostly nuclear. | That compartment conflicts with the reported membrane topology and cytoplasmic or membranous tissue pattern (UniProt P78504 topology; HPA tissue IHC profile). | Review the negative control, antibody conditions, and chromogen distribution before scoring; document any remaining nuclear signal as discordant (general IHC practice). |
| Bone-marrow hematopoietic cells stain strongly. | HPA reports JAG1 as not detected in that cell population; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC: Not detected; general IHC practice). | Inspect appropriate negative and detection controls, then assess signal in the identified cells separately from other marrow cells (general IHC practice). |
| The whole section has diffuse background. | Nonspecific staining or detection background can obscure the cell-specific HPA pattern (general IHC practice; HPA tissue IHC). | Compare with a negative control and review blocking, washing, and detection conditions before changing the biological interpretation (general IHC practice). |
| Only weak signal appears in cerebral-cortex endothelial or lung alveolar cells. | HPA reports Low staining in those cell populations, so weak signal may fit the reference pattern (HPA tissue IHC: Low). | Identify the relevant cells and compare them with a reported high-staining reference and run controls before calling the assay failed (HPA tissue IHC: High; general IHC practice). |
| Cytoplasmic staining is present without a sharp membrane outline. | HPA describes both cytoplasmic and membranous tissue staining, so cytoplasmic signal alone is not automatically discordant (HPA tissue IHC profile). | Confirm that staining follows the expected cell populations and compare background controls; avoid requiring an exclusively membrane-rim pattern (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing JAG1 staining
M00640-1 · IHCImmunohistochemical analysis of paraffin-embedded Rat liver, using the Antibody at 1:200 dilution.
M00640-1 · IHCImmunohistochemical analysis of paraffin-embedded Human lung large cell cancer, using the Antibody at 1:50 dilution.
M00640-1 · IFImmunofluorescent analysis using the Antibody at 1:50 dilution.
A00640 · IHCImmunohistochemistry of JAG1 in human spleen tissue with JAG1 antibody at 5 μg/ml.
A00640 · IFImmunofluorescence of JAG1 in human spleen tissue with JAG1 antibody at 20 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000101384-JAG1/tissue/cerebral+cortex#imid_2865384
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000101384-JAG1/tissue/colon#imid_2865326
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000101384-JAG1/tissue/liver#imid_2865319
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000101384-JAG1/tissue/gallbladder#imid_2865303
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB010343; green = JAG1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB010343; green = JAG1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for JAG1 IHC & IF
🧪Run gallbladder first; its glandular cells should stain (HPA: High in gallbladder glandular cells). Use bone marrow hematopoietic cells as the negative tissue (HPA: Not detected in bone marrow hematopoietic cells); on the gallbladder slide, cells without specific staining should show only counterstain or background, without assigning them a known JAG1-negative identity.
Positive control tissue: Gallbladder (Glandular cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show JAG1 in A-431, U-251MG, U2OS, with annotated localisation: Golgi apparatus (approved), Plasma membrane (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched isotype control matched to the primary antibody’s host species and subclass, and JAG1-knockout tissue as a biological negative (standard IHC controls). For chromogenic detection, block endogenous peroxidase and check gallbladder background; block endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; optimize retrieval on matched sections (supplied evidence; standard IHC practice). The selected catalog antibody caption shows paraffin-section staining of rat liver at 1:200, but its fixative is unreported (caption: M00640-1). The supplied evidence does not establish that frozen sections or IF are easier, or identify a gallbladder-specific artefact; assess background against the controls (supplied evidence; standard IHC practice).
HPA tissue IHC evidence for JAG1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Skin | Eccrine glands | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced JAG1 IHC Tips
Use compartment, cell type, and matched controls to troubleshoot JAG1 staining in paraffin sections (UniProt P78504 topology; HPA tissue IHC).
How should I optimize retrieval when JAG1 staining is weak in paraffin sections?
Start with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval setting). Allow sections to cool in retrieval buffer before washing, and keep heating and cooling consistent across the comparison set (standard IHC practice). Use a glandular-cell positive control, such as gallbladder, to judge whether weak staining reflects the run or the specimen (HPA: high in gallbladder glandular cells). If signal remains weak, vary retrieval time in a small controlled series while holding antibody concentration and detection constant (standard IHC practice). Assess membrane and cytoplasmic signal separately because both appear in tissue IHC (HPA: ubiquitous cytoplasmic and membranous expression).
Could fixation explain variable JAG1 staining between my paraffin blocks?
The selected rat-liver paraffin-section caption does not state a fixative, and target-specific JAG1 fixation sensitivity is unknown from the supplied evidence (caption: M00640-1). Record each block’s fixative, fixation interval, processing history, and section age before comparing staining intensity (standard IHC practice). Run blocks with known processing histories together, using the same retrieval, antibody incubation, and chromogenic development conditions (standard IHC practice). If one block is weak, repeat it alongside a positive-control section before changing retrieval or antibody concentration (standard IHC practice; HPA: high in gallbladder glandular cells). Do not assign a fixation effect from a single negative section without comparable controls (standard IHC practice).
Should JAG1 staining appear at the cell surface or in the cytoplasm?
Score membrane staining explicitly: JAG1 has an extracellular region at residues 34–1067, one transmembrane segment at 1068–1093, and a cytoplasmic tail at 1094–1218 (UniProt P78504 topology). Cytoplasmic signal can also be plausible in tissue sections, where the reported profile is cytoplasmic and membranous (HPA: tissue IHC profile). Cell-based localisation data place JAG1 at the plasma membrane and Golgi apparatus (HPA: approved subcellular locations). Compare staining within the same identified cell population and inspect the membrane pattern at high magnification (standard IHC practice). Predominantly nuclear staining requires control-based scrutiny because the supplied localisation evidence does not place JAG1 in nuclei (UniProt P78504 subcellular; HPA subcellular).
How can an unknown antibody epitope affect JAG1 staining interpretation?
JAG1 has 2 recorded isoforms, but the supplied product caption does not identify this antibody’s epitope or establish which isoforms it recognizes (UniProt P78504 isoforms; caption: M00640-1). The extracellular region contains DSL and EGF-like domains, while the cytoplasmic tail follows the transmembrane segment (UniProt P78504 domains and topology). Nine annotated glycosylation sites occur in the extracellular region; whether they affect this antibody’s staining is untested here (UniProt P78504 glycosylation; supplied product evidence). Seek an epitope map before attributing compartment differences or negative cells to a particular isoform (standard IHC interpretation practice). Compare any epitope-dependent hypothesis against matched positive and negative controls (standard IHC practice).
How should I investigate JAG1 localisation with a separate IF assay?
For the separate IF/ICC assay, pair JAG1 with a marker identifying the expected cell population, such as an epithelial marker in gallbladder glands (HPA: high in gallbladder glandular cells; standard IF practice). Select a spectrally separated, preferably far-red JAG1 fluorophore after checking tissue autofluorescence and single-stain controls (standard IF practice). An extracellular epitope may be accessible without permeabilisation, whereas a cytoplasmic-tail epitope requires membrane access; the catalog antibody’s epitope is unspecified (UniProt P78504 topology; caption: M00640-1). Compare permeabilised and unpermeabilised conditions while keeping imaging settings fixed (standard IF practice). No directly relevant IF fixation evidence is supplied, so establish fixation conditions with assay controls (supplied evidence; standard IF practice).
What should I check when JAG1 chromogenic staining looks diffuse?
Check a no-primary control and inspect whether colour follows tissue structures or appears over the whole section (standard IHC practice). Block endogenous peroxidase before HRP and DAB detection, then compare background under identical development conditions (standard chromogenic IHC practice). Reduce nonspecific binding with an appropriate protein block and titrate the primary antibody if background persists (standard IHC practice). The selected antibody was pictured on paraffin-embedded rat liver at 1:200; that caption does not establish the best dilution for other specimens (caption: M00640-1). Judge any remaining diffuse signal against the reported cytoplasmic and membranous pattern within identifiable cells (HPA: tissue IHC profile).
How should I quantify JAG1 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring compartment before reviewing outcomes, because tissue IHC reports both cytoplasmic and membranous JAG1 staining (HPA: tissue IHC profile; standard scoring practice). For intensity scoring, calculate an H-score from the percentages at 0, 1+, 2+, and 3+ intensity, yielding 0–300 (standard IHC scoring practice). Alternatively, report percentage-positive cells or positive-cell density per mm² within a prespecified tissue region (standard IHC scoring practice). Normalize counts to the number of evaluable cells or measured viable area, and report compartments separately (standard IHC scoring practice). Hold retrieval, detection, development, and image thresholds constant across specimens (standard IHC practice).
How can I distinguish convincing JAG1 positivity from staining artefacts?
Convincing staining should occur in identifiable cells with membrane or cytoplasmic signal, consistent with the tissue IHC profile (HPA: ubiquitous cytoplasmic and membranous expression). Check whether the relevant cell type matches the reference: gallbladder glandular cells stain highly, while bone-marrow hematopoietic cells were not detected in the cited profile (HPA: gallbladder and bone marrow tissue IHC). Exclude section-edge accentuation and necrotic regions before calling a specimen positive (standard IHC interpretation practice). Investigate nuclear-only or structureless deposits against the no-primary control and expected JAG1 localisation (UniProt P78504 subcellular; standard IHC practice). Persistent colour in the control can indicate endogenous enzyme activity or nonspecific detection (standard chromogenic IHC practice).
Boster reagents
Best JAG1 / Protein jagged-1 IHC Antibodies
IHC images show JAG1 staining in human spleen and paraffin-embedded rat liver (A00640 image caption; M00640-1 image caption); IF imagery shows human spleen (A00640 image caption).
Real IHC data
Anti-JAG1 Rabbit Monoclonal Antibody
Cat # M00640-1
Real IHC data
Anti-Protein jagged-1 JAG1 Antibody
Cat # A00640
M00640-1 will render with a paraffin-embedded rat liver IHC image, although its listed reactivity is Human (M00640-1 image caption; catalog reactivity). A00640 will render with a human spleen IHC image; its applications include IHC-P and IF, with Human, Mouse and Rat listed as reactive species (A00640 image caption; catalog applications and reactivity).
Which to pick: For paraffin-section tissue IHC, M00640-1 has a paraffin-section image and is a rabbit monoclonal; its fixative is unreported, and its listed reactivity is Human despite the rat liver caption (M00640-1 image caption; catalog host, clone and reactivity). Choose A00640 for IF or a cross-species starting point because IF is listed and Human, Mouse and Rat are listed as reactive; its human spleen IHC caption does not report the fixative, and ICC is not listed (A00640 catalog applications and reactivity; A00640 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P78504 (JAG1_HUMAN, Protein jagged-1).
- Human Protein Atlas. JAG1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. JAG1 subcellular location (ICC-IF): Localized to the plasma membrane and the Golgi apparatus..
- Human Protein Atlas. JAG1 antibody validation summary (2 antibodies).
- High expression of the Notch ligand Jagged-1 is associated with poor prognosis after surgery for colorectal cancer. Cancer science 2016 — PMC5132269.
- Jagged-1 induced molecular alterations in HPV associated invasive squamous cell and adenocarcinoma of the human uterine cervix. Scientific reports 2018 — PMC6008329.
- Jagged 1 is a major Notch ligand along cholangiocarcinoma development in mice and humans. Oncogenesis 2016 — PMC5177771.
- Jagged1 Instructs Macrophage Differentiation in Leprosy. PLoS pathogens 2016 — PMC4988718.
- PubMed PMID:9268641 — UniProt-cited evidence.
- PubMed PMID:9207788 — UniProt-cited evidence.
- PubMed PMID:9462510 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for JAG1 (Protein jagged-1). Expected localisation Cytoplasmic and membranous in tissue (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: M00640-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Gallbladder (Glandular cells); negative Bone marrow. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Rat liver tissue; fixative not specified (datasheet M00640-1); 2. Fixation: Image fixative and duration unreported (datasheet M00640-1); 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 citrate buffer, pH 6.0 , 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 monoclonal (clone 17J22) anti-JAG1, 1:50-1:200 (datasheet M00640-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Antibody staining has medium consistency with RNA (HPA tissue IHC); Isoform / epitope: 2 isoforms; map extracellular vs cytoplasmic epitopes (UniProt); Regulation: Higher in cervical squamous carcinoma (UniProt). Evidence: UniProt P78504, HPA tissue IHC · Supported, HPA ICC-IF · plasma membrane and the Golgi apparatus., Datasheet M00640-1, PMC5132269, PMC5177771. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/JAG1/immunohistochemistry-ihc-guide.