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CD1B / T-cell surface glycoprotein CD1b · IHC design guide
Design Immunohistochemistry for CD1B
Plan CD1B chromogenic IHC in paraffin sections using the catalog antibody’s documented tonsil protocol (datasheet A02158-1). Assess lysosomal staining with appropriate controls, bearing in mind its low consistency with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CD1B 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 | Lysosomal staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Lysosomal staining in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A02158-1) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Bone marrow+4 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 | Staining shows low consistency with RNA (HPA tissue IHC) | |
| Regulation | Expression regulation is not specified (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unknown; check sidedness (UniProt) |
Section 1
Recommended CD1B IHC & IF Protocols
Compare the catalog antibody’s citrate pH 6 HIER protocol (datasheet A02158-1) with the published CD1B IHC protocols listed below (PMC7055113; PMC6553556; PMC13553427; PMC11629625).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A02158-1) |
| Fixation | Image fixative and duration unreported (datasheet A02158-1); 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 A02158-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02158-1) |
| Primary antibody | Rabbit anti-CD1B, 1μg/ml (datasheet A02158-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02158-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02158-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CD1B-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Lysosomal expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6 HIER for the catalog antibody (datasheet A02158-1). The sheep protocol used pronase digestion for CD1b retrieval (PMC7055113).
Section 2
What Is the Expected CD1B Staining Pattern?
CD1B can appear at the cell surface and in intracellular compartments as it traffics through endosomes and lysosomes (UniProt P29016 topology and subcellular location). In paraffin-section IHC, HPA reports lysosomal staining in several tissues, including medium staining in adipocytes and selected glandular cells (HPA tissue IHC). Treat these as observed patterns, not universal expectations: HPA rates its IHC profile Approved but reports low consistency with RNA expression (HPA tissue IHC).
| Discrete intracellular granules or a vesicular pattern in adipocytes or glandular cells, with possible cell-edge staining. | This is compatible with HPA’s lysosomal IHC profile and its medium staining in adipocytes and selected glandular cells (HPA tissue IHC). A cell-edge component is biologically plausible because CD1B has an extracellular domain and one transmembrane segment (UniProt P29016 topology). Score the compartments separately. |
| Strong, uniform nuclear staining with little cytoplasmic or membrane-associated signal. | A nuclear-dominant pattern conflicts with the reported membrane, endosome and lysosome locations (UniProt P29016 subcellular location) and HPA’s lysosomal IHC profile (HPA tissue IHC). Consider nonspecific staining or an imaging/scoring error; this pattern alone does not establish CD1B positivity. |
| Prominent staining in skeletal-muscle myocytes or bone-marrow hematopoietic cells. | HPA reports CD1B as not detected in those specific cell populations (HPA tissue IHC). Recheck cell identity and compare controls; unexpected signal could reflect cross-reactivity or endogenous chromogen-generating activity (general IHC practice). A different staining result does not, by itself, identify the cause. |
| Diffuse chromogen across cells, stroma and slide background, without a defined cellular compartment. | This does not resemble the reported lysosomal tissue pattern (HPA tissue IHC). Inspect the no-primary control and review blocking, washes and detection chemistry for background (general IHC practice). Diffuse signal cannot be scored as CD1B solely because the section contains an HPA-reported positive cell type. |
| No staining in an adipocyte or glandular-cell population selected as a positive comparator. | HPA reports medium staining in adipocytes from adipose tissue or breast and in selected glandular cells (HPA tissue IHC). Verify that the relevant cells are present, then assess the antibody and detection run with controls (general IHC practice). HPA’s Approved rating and low RNA–staining consistency limit how strongly one negative section can be interpreted (HPA tissue IHC). |
💡Expected CD1B appearanceCall positive when a relevant cell population shows discernible vesicular or lysosomal staining, potentially with a cell-edge component, around HPA’s reported medium level; nuclear-only or diffuse background is suspect (HPA tissue IHC; UniProt P29016 subcellular location and topology).
| Compartment and topology | CD1B spans the membrane at residues 304–324, with residues 18–303 extracellular and 325–333 cytoplasmic (UniProt P29016 topology). Its trafficking between the surface, endosomes and lysosomes permits both cell-edge and intracellular staining (UniProt P29016 subcellular location). The record does not locate the catalog antibody’s epitope. |
| Choice of tissue and cell population | HPA reports medium staining in adipocytes and several glandular-cell populations, but no detection in skeletal-muscle myocytes and bone-marrow hematopoietic cells (HPA tissue IHC). Use the named cell population when comparing sections; a whole-tissue label can conceal mixed cells and focal staining (general IHC interpretation). |
| Strength of the tissue evidence | The tissue profile is Approved, yet HPA explicitly notes low consistency between antibody staining and RNA expression; the listed antibody HPA021824 is IHC Approved, without an IHC Enhanced designation in the supplied record (HPA tissue IHC; HPA antibody validation). Interpret an unexpected distribution cautiously and retain controls. |
| Antigen retrieval and detection | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| Situation | Likely cause | Next action |
|---|---|---|
| A presumed positive section has no chromogen. | The expected cell population may be absent, or the IHC run may have failed (general IHC practice); HPA’s positive calls apply to named cells and staining levels (HPA tissue IHC). | Locate the named cells, inspect a run control and confirm primary-antibody and detection steps (general IHC practice). Avoid calling the tissue CD1B-negative until the run is valid. |
| The no-primary control is stained. | Endogenous detection activity or nonspecific chromogen deposition is possible (general IHC practice). This control cannot demonstrate CD1B binding. | Address the relevant endogenous activity, blocking or wash issue, then repeat the control alongside the primary-antibody section (general IHC practice). |
| Signal fills nuclei or appears uniformly across unrelated cell types. | The pattern conflicts with CD1B’s reported membrane and intracellular trafficking locations (UniProt P29016) and HPA’s lysosomal IHC profile (HPA tissue IHC). | Check counterstain, focus, control sections and background before scoring; require a defined cellular compartment in the named cell population (general IHC practice). |
| Skeletal-muscle myocytes or bone-marrow hematopoietic cells stain prominently. | Those cell populations are listed as not detected (HPA tissue IHC); possible explanations include mistaken cell identification, nonspecific antibody binding or detection background (general IHC practice). | Confirm the stained cell type and compare no-primary and tissue controls. Report the unexpected pattern instead of treating it as established CD1B expression. |
| A positive cell population shows only faint or scattered granules. | HPA’s medium staining examples describe observed levels, not a guaranteed intensity for every section (HPA tissue IHC). Section quality and run conditions can complicate interpretation (general IHC practice). | Score the fraction of relevant cells, intensity and compartment together; compare the same run’s controls and document uncertainty when the pattern is below a clear positive call (general IHC practice). |
| Q: Should ICC-IF Golgi or vesicle staining dictate the paraffin-section IHC call? | HPA’s ICC-IF summary places CD1B mainly in Golgi apparatus and vesicles, with additional cytosol, whereas its tissue IHC summary describes lysosomal expression (HPA subcellular ICC-IF; HPA tissue IHC). | A: Use the tissue IHC pattern and cell population for this chromogenic section. Treat the ICC-IF observations as context, not an IHC protocol or a requirement to assign every IHC granule to the Golgi. |
Experiment evidence
IHC & IF images showing CD1B staining
A02158-1 · IHCIHC analysis of CD1b using anti-CD1b antibody (A02158-1). CD1b was detected in paraffin-embedded section of human tonsil tissue. Heat mediat
A02158-1 · IHCIHC analysis of CD1b using anti-CD1b antibody (A02158-1). CD1b was detected in paraffin-embedded section of human tonsil tissue. Heat mediat
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000158485-CD1B/tissue/cerebral+cortex#imid_5834644
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000158485-CD1B/tissue/colon#imid_5834570
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000158485-CD1B/tissue/liver#imid_5834563
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000158485-CD1B/tissue/kidney#imid_5834556
HPA · ICC-IF JURKATHuman Protein Atlas immunofluorescence, JURKAT, antibody HPA021824; green = CD1B, blue = nucleus, red = microtubules.
HPA · ICC-IF JURKATHuman Protein Atlas immunofluorescence, JURKAT, antibody HPA021824; green = CD1B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CD1B IHC & IF
🧪Run adipose tissue first: adipocytes should stain (HPA: Medium in adipocytes). Use skeletal muscle myocytes as the negative tissue (HPA: Not detected in myocytes); non-adipocyte cells on the adipose slide should show only background where they lack specific staining, but their CD1B-negative status requires independent validation.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CD1B in JURKAT, U2OS, with annotated localisation: Golgi apparatus (approved), Vesicles (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls, plus a CD1B knockout biological negative (selected tissue-IHC caption: rabbit anti-CD1b antibody). Quench endogenous peroxidase and assess endogenous biotin background because the selected tissue-IHC caption uses avidin–biotin detection with DAB.
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A02158-1 paraffin-section caption does not state the fixative (selected tissue-IHC caption: fixative not stated). The reported IHC procedure uses citrate retrieval at pH 6 for 20 minutes (selected tissue-IHC caption); whether frozen sections or IF are easier is unreported. Adipose sections can lose lipid during processing, leaving empty spaces that should not be scored as staining (standard histology practice).
HPA tissue IHC evidence for CD1B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CD1B IHC Tips
Troubleshoot CD1B staining in paraffin sections by checking retrieval, compartment, cell identity and controls before interpreting chromogenic signal.
How should I adjust retrieval when CD1B staining is weak or uneven?
Begin with heat-mediated citrate retrieval at pH 6 for 20 minutes in paraffin sections (datasheet A02158-1). Run a known positive section alongside the test section, and keep heating, cooling and wash conditions consistent so differences are attributable to the specimen (standard IHC practice). If staining remains weak, compare a modestly longer retrieval time on adjacent sections while checking whether tissue morphology or background deteriorates (standard IHC practice). Record both signal intensity and its location: CD1B traffics among the surface, endosomes and lysosomes, so a change in compartmental staining needs assessment before accepting a stronger signal (UniProt P29016 localisation).
Could fixation explain a negative CD1B result in paraffin sections?
The selected paraffin-section caption does not report its fixative, so target-specific CD1B fixation sensitivity is unknown (datasheet A02158-1). Confirm the specimen's recorded fixative and fixation duration, then compare it with a concurrently processed positive control before attributing absent staining to antigen loss (standard IHC practice). Keep section thickness, citrate retrieval at pH 6 and detection conditions matched during that comparison (datasheet A02158-1; standard IHC practice). If the control stains but the test section does not, examine tissue preservation, cell identity and reagent controls; a negative result alone cannot establish a CD1B-specific fixation effect (standard IHC practice).
Should CD1B appear at the surface or inside cells?
Assess both membranous and punctate intracellular DAB signal: CD1B traffics between the cell membrane, endosomes and lysosomes (UniProt P29016 localisation). Its extracellular region spans residues 18–303, followed by a transmembrane segment at 304–324 and a short cytoplasmic tail at 325–333 (UniProt P29016 topology). HPA describes lysosomal tissue staining, while its cellular imaging places signal mainly in Golgi apparatus and vesicles, with additional cytosolic signal (HPA tissue IHC; HPA subcellular). Compare the observed pattern with a positive control and tissue morphology; diffuse DAB across damaged areas needs scrutiny before it is scored as CD1B (standard IHC practice).
Can I interpret different staining patterns as CD1B isoforms?
CD1B has 2 listed isoforms, but the supplied evidence does not map this antibody's epitope to either one (UniProt P29016 isoforms; datasheet A02158-1). Check the antibody's documented immunogen or epitope before claiming isoform selectivity, and report staining as CD1B when that information is unavailable (standard IHC practice). CD1B contains an Ig-like region at residues 185–295 and four listed glycosylation sites, yet these annotations do not establish which feature this antibody recognizes (UniProt P29016 domains and glycosylation). Compare adjacent sections under identical retrieval and detection conditions before interpreting a pattern difference as biological rather than technical (standard IHC practice).
How can IF help assess ambiguous CD1B staining in tissue?
Use IF as a complementary localisation check, while evaluating it separately from the catalog antibody's paraffin-section chromogenic result (datasheet A02158-1; standard IF practice). Multiplex CD1B with a marker for the expected cell population, such as CD3 when examining cortical thymocytes, and include a nuclear counterstain to identify individual cells (UniProt P29016 tissue specificity; standard IF practice). Choose fluorophores after inspecting an unstained section for autofluorescence, reserving a spectrally distinct channel for a weak CD1B signal (standard IF practice). For an extracellular epitope, assess surface staining before permeabilisation; for an intracellular epitope, compare controlled permeabilisation because CD1B also occupies endosomal and lysosomal membranes (UniProt P29016 topology and localisation; standard IF practice).
What should I check when DAB obscures CD1B staining?
Inspect a no-primary control and the section margins first; widespread color in those controls or damaged regions argues against assigning every deposit to CD1B (standard IHC practice). In a biotin-based DAB workflow, assess endogenous peroxidase and biotin contributions with appropriate controls, since the selected caption uses a biotinylated secondary and avidin–biotin detection (datasheet A02158-1; standard IHC practice). The caption used 10% goat serum blocking and 1 µg/ml primary antibody overnight at 4°C; use those documented conditions as a comparison point (datasheet A02158-1). If background persists, titrate primary antibody and detection exposure on adjacent sections while preserving a positive control (standard IHC practice).
How should I quantify heterogeneous CD1B IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because CD1B can occur at the surface and in intracellular trafficking compartments (UniProt P29016 localisation; standard IHC practice). Report the percentage of positive target cells and staining intensity, or calculate an H-score using intensity categories 0–3 and their respective cell percentages (standard IHC practice). For focal populations, positive-cell density per mm² can be useful; normalise it to the measured viable tissue area and report the counted cell population (standard IHC practice). Keep retrieval, illumination, chromogen development and scoring thresholds consistent, and exclude folds or necrotic areas from the denominator (standard IHC practice).
How do I distinguish genuine CD1B staining from artefact?
Seek reproducible staining in intact cells with membranous or punctate intracellular distribution, consistent with CD1B trafficking through the cell surface, endosomes and lysosomes (UniProt P29016 localisation). Check cell identity rather than treating every brown cell as expected: UniProt reports cortical thymocytes, while HPA lists several other stained cell populations and notes low agreement between staining and RNA data (UniProt P29016 tissue specificity; HPA tissue IHC). Discount staining confined to section edges, necrosis or the no-primary control, and investigate endogenous enzyme signal before assigning positivity (standard IHC practice). Interpret intensity against a concurrently stained positive control, with uncertain patterns recorded as indeterminate (standard IHC practice).
Boster reagents
Best CD1B / T-cell surface glycoprotein CD1b IHC Antibodies
One human-reactive antibody has real IHC data from a paraffin-embedded tonsil section (catalog image: A02158-1). No IF/ICC data are provided (catalog: applications, image alts).
Real IHC data
Anti-CD1b Antibody ®
Cat # A02158-1
A02158-1 is listed for human IHC (catalog: applications, reactivity). Its IHC image shows CD1b staining in a paraffin-embedded human tonsil section (catalog image: A02158-1).
Which to pick: Choose A02158-1 for human paraffin-section IHC: it is rabbit polyclonal (catalog: host, dilution_raw), and its own image documents staining after citrate retrieval (catalog image: A02158-1). The fixative is unreported (catalog image: A02158-1). No listed SKU has IF/ICC validation or nonhuman reactivity, so neither use has a supported pick here (catalog: applications, reactivity, IF image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P29016 (CD1B_HUMAN, T-cell surface glycoprotein CD1b).
- Human Protein Atlas. CD1B tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CD1B subcellular location (ICC-IF): Mainly localized to the Golgi apparatus and vesicles. In addition localized to the cytosol..
- Human Protein Atlas. CD1B antibody validation summary (1 antibodies).
- Characterization of dendritic cells and follicular dendritic cells in the hepatic lymph nodes and liver of sheep experimentally infected with Fasciola hepatica. Veterinary research 2020 — PMC7055113.
- Vitamin A Metabolism by Dendritic Cells Triggers an Antimicrobial Response against Mycobacterium tuberculosis. mSphere 2019 — PMC6553556.
- Identifying dendritic cell heterogeneity and potential risk genes in atopic dermatitis: integrative scRNA-seq, bulk RNA-seq analyses and experimental validation. Frontiers in medicine 2026 — PMC13553427.
- Establishment of CD1b-restricted immunity to lipid antigens in the pulmonary response to Mycobacterium tuberculosis infection. Infection and immunity 2024 — PMC11629625.
- PubMed PMID:2447586 — UniProt-cited evidence.
- PubMed PMID:2701945 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CD1B (T-cell surface glycoprotein CD1b). Expected localisation Lysosomal staining in several tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A02158-1). Validated IHC antibody: A02158-1 at 1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Bone marrow. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A02158-1); 2. Fixation: Image fixative and duration unreported (datasheet A02158-1); 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 retrieval: Citrate pH 6 , 20 min (datasheet A02158-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02158-1); 8. Primary antibody: Rabbit anti-CD1B, 1μg/ml (datasheet A02158-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02158-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining shows low consistency with RNA (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unknown; check sidedness (UniProt); Regulation: Expression regulation is not specified (UniProt). Evidence: UniProt P29016, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the Golgi apparatus and vesicles. In addition…, Datasheet A02158-1, PMC7055113, PMC6553556. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CD1B/immunohistochemistry-ihc-guide.