CD1B / T-cell surface glycoprotein CD1b · IHC design guide

Design Immunohistochemistry for CD1B

Plan chromogenic CD1B IHC in paraffin sections using the catalog antibody’s 0.5–1 µg/mL range and documented tonsil staining (datasheet A02158-1). Assess the lysosomal tissue pattern while accounting for low consistency between antibody staining and RNA expression (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for CD1B (IHC for CD1B): expected localisation Lysosomal tissue staining (HPA tissue IHC); membrane trafficking expected (UniProt), antibody A02158-1, validated IHC image, and IHC protocol steps — Printable **CD1B** IHC protocol sheet — expected localisation Lysosomal tissue staining (HPA tissue IHC); membrane trafficking expected (UniProt), antibody A02158-1, controls and protocol steps. Open the full CD1B IHC guide →

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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 tissue staining (HPA tissue IHC); membrane trafficking expected (UniProt)
	Staining pattern	Lysosomal staining in several tissues; glandular cells stain (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 (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02158-1)
	Caveat	Antibody staining and RNA show low consistency (HPA tissue IHC)
	Regulation	Lymphoid tissue-enriched RNA (HPA tissue RNA)
	Isoform / epitope	2 isoforms; epitope differences unspecified. Check extracellular versus cytoplasmic location (UniProt)

Section 1

Recommended CD1B IHC & IF Protocols

Compare the catalog antibody’s citrate pH 6 heat retrieval protocol (datasheet A02158-1) with three published CD1B IHC protocols (PMC7055113; PMC6553556; PMC13553427).

Recommended immunohistochemistry (IHC-P) protocol parameters
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 heat retrieval for the catalog antibody (datasheet A02158-1). The sheep study used pronase digestion for its CD1b antibody (PMC7055113).

Section 2

What Is the Expected CD1B Staining Pattern?

CD1B can occupy the cell surface, endosomes and lysosomes (UniProt P29016: localization and topology). In paraffin IHC, expect chiefly lysosomal staining in the cell populations reported by HPA, including adipocytes and glandular or respiratory epithelial cells in selected tissues (HPA: tissue IHC). HPA rates the tissue profile Approved but reports low consistency between antibody staining and RNA expression; interpret staining with that limitation (HPA: reliability).

What am I looking at on my slide?
Granular intracellular staining in colon glandular cells or adipocytes; signal may also outline a cell membrane.	This fits HPA's lysosomal profile and Medium staining in colon glandular cells and adipocytes (HPA: tissue IHC). A membrane component is biologically plausible because CD1B traffics through the cell surface, endosomes and lysosomes (UniProt P29016: localization). Score the identified cell population and compartment together, rather than calling any brown deposit positive.
Strong nuclear staining, or staining confined to an extracellular deposit, without a credible cellular pattern.	Neither location is among CD1B's reported cell membrane, endosome and lysosome locations (UniProt P29016: localization). Treat that pattern as suspect for artefact and inspect morphology, detection controls and the antibody's staining distribution before assigning CD1B positivity (general IHC practice).
Prominent staining in bone marrow hematopoietic cells or skeletal muscle myocytes.	Those specific populations are Not detected in the supplied tissue IHC observations (HPA: tissue IHC). Consider cross-reactivity or endogenous detection activity, while allowing for sample and assay differences (general IHC practice). The HPA observations concern named cells; they do not establish that every cell in either tissue is CD1B negative.
A uniform haze covers positive cells, neighboring cells and empty slide areas.	A field-wide haze obscures the cell-restricted, chiefly lysosomal pattern described for CD1B (HPA: tissue IHC). Assess background with an appropriate reagent control, and review blocking, washing and detection conditions before scoring intensity (general IHC practice). Haze alone cannot identify which workflow step caused it.
No convincing signal in colon glandular cells or adipocytes on an otherwise interpretable section.	HPA reports Medium staining for those populations, making them useful comparison sites rather than guaranteed positive controls (HPA: tissue IHC). Check section integrity, antigen retrieval, antibody and detection performance with suitable controls (general IHC practice). A single negative section does not overturn the reported tissue profile.

💡Expected CD1B appearanceCall a result consistent with CD1B when selected HPA-reported cells show interpretable, chiefly granular lysosomal staining around the reported Medium level; a membrane component can fit CD1B trafficking, whereas strong nuclear or indiscriminate field-wide staining is suspect (HPA: tissue IHC; UniProt P29016: localization; general IHC practice).

How each factor affects the staining
Trafficking and compartment	CD1B moves among the cell surface, endosomes and lysosomes (UniProt P29016: localization). HPA describes lysosomal tissue staining (HPA: tissue IHC). Read a mixed membrane and intracellular pattern in that context; a nuclear pattern lacks support from either source.
Epitope position	The mature chain spans residues 18–333; residues 18–303 are extracellular and 304–324 form the transmembrane segment (UniProt P29016: processing and topology). These facts cannot establish where an unspecified antibody binds or predict its paraffin IHC performance.
Tissue and cell context	HPA reports Medium staining in selected adipocytes, glandular cells and respiratory epithelial cells, but no detection in specified bone marrow hematopoietic cells or myocytes (HPA: tissue IHC). Compare the same named cell populations; whole-tissue labels can hide that distinction.
Strength of the tissue evidence	The listed antibody's IHC status is Approved, and HPA reports low consistency between antibody staining and RNA expression (HPA: antibody validation and tissue reliability). Use morphology and controls when interpreting unexpected sites; Approved does not mean every reported cell pattern is independently confirmed.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected cells are unstained.	The assay or section may have failed, or this specimen may differ from HPA's observed examples (HPA: tissue IHC; general IHC practice).	Compare a suitable HPA-reported Medium population and run detection controls; review retrieval and antibody conditions as general IHC checks, without assuming a CD1B-specific fixation effect.
DAB signal is strong in an HPA-listed negative cell population.	Cross-reactivity or endogenous peroxidase can mimic a positive stain (general IHC practice); the named population is Not detected by HPA (HPA: tissue IHC).	Inspect the exact cell identity and run appropriate antibody-omission and detection controls (general IHC practice).
Signal appears predominantly nuclear.	Nuclear localization is absent from the supplied CD1B locations (UniProt P29016: localization; HPA: tissue IHC).	Treat the pattern as suspect; review counterstain, tissue morphology and control slides before scoring it (general IHC practice).
Granules cannot be distinguished from widespread background.	Background can mask compartment-specific staining (general IHC practice); HPA describes lysosomal tissue expression (HPA: tissue IHC).	Check reagent controls and review blocking, washing and detection settings; score only signal tied to identifiable cells (general IHC practice).
Only membrane staining is visible in a candidate positive cell.	Surface localization is plausible, but HPA's tissue summary emphasizes lysosomes (UniProt P29016: localization; HPA: tissue IHC).	Confirm the cell population and compare morphology and controls before deciding whether the membrane signal is credible (general IHC practice).
What if IF/ICC shows Golgi or vesicular signal instead of the IHC pattern?	HPA reports approved Golgi and vesicle locations, plus cytosol as an additional location, in ICC-IF (HPA: subcellular ICC-IF).	Interpret that result within its separate IF/ICC assay and controls; do not use it alone to redefine the lysosomal tissue IHC profile (HPA: subcellular ICC-IF and tissue IHC).

Source: UniProt P29016 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing CD1B staining

IHC analysis of CD1b using anti-CD1b antibody (A02158-1). CD1b was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CD1b Antibody (A02158-1) 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.

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.

← swipe / scroll for more images →

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

Sample controls for CD1B IHC & IF

🧪Run bronchus first: respiratory epithelial cells are reported at Medium CD1B staining (HPA: bronchus). Use skeletal muscle myocytes as the negative tissue because CD1B is not detected there (HPA: skeletal muscle); on the bronchus slide, internal negative cells should lack specific chromogen staining, but verify their identity and signal before treating them as negative controls (standard IHC practice).

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 a no-primary (secondary-only) control, a concentration-matched rabbit IgG isotype control, and, if available, a CD1B knockout biological control (caption: rabbit anti-CD1b antibody; standard IHC practice). Block endogenous peroxidase and assess background in inflammatory cells on bronchus sections before interpreting DAB signal (caption: DAB detection; standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02158-1 tissue-IHC caption does not state the fixative (caption: fixative not stated). The paraffin-section example uses heat-mediated citrate retrieval at pH 6 for 20 minutes, so start with that condition while checking retrieval dependence experimentally (caption: citrate retrieval). The supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC; bronchial inflammatory cells can contribute endogenous-peroxidase background to chromogenic detection (caption: paraffin IHC with DAB; standard IHC 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

Use the catalog antibody’s paraffin-section protocol as the starting point, then assess CD1B staining by compartment, morphology, and controls.

What retrieval should I use if CD1B staining is weak in paraffin sections?

Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A02158-1). Keep heating and cooling conditions consistent between sections so a change in signal can be attributed to the antibody or tissue rather than retrieval (standard IHC practice). The catalog image used paraffin-embedded human tonsil, followed by 1 μg/ml primary antibody overnight at 4°C (datasheet A02158-1). If staining remains weak, check section adhesion, antibody incubation, and detection controls before adjusting retrieval time on matched sections; prolonged heating can damage morphology (standard IHC practice).

How should I assess fixation effects on CD1B IHC staining?

The selected paraffin-section caption does not state a fixative, so CD1B-specific fixation sensitivity is unknown for this antibody (datasheet A02158-1). Record the fixative, fixation duration, processing history, and section age for every specimen before comparing staining (standard IHC practice). Compare similarly processed positive-control sections alongside test sections, using the same citrate pH 6 retrieval for 20 minutes (datasheet A02158-1). If signal differs across processing batches, repeat staining with matched controls and inspect morphology; do not attribute the difference to a CD1B-specific fixation effect without direct evidence (standard IHC practice).

Should CD1B stain the cell surface or intracellular structures?

Assess membrane staining and intracellular puncta separately: CD1B traffics among the cell membrane, endosomes, and lysosomes (UniProt P29016 localisation). Its extracellular region spans residues 18–303, its transmembrane segment spans 304–324, and its cytoplasmic tail spans 325–333 (UniProt P29016 topology). Tissue IHC has been described as lysosomal, while cell imaging reports Golgi and vesicular localisation with additional cytosolic signal (HPA tissue IHC; HPA subcellular). Use cell boundaries, morphology, and a counterstain to distinguish true membranous or punctate staining from diffuse chromogen; compare compartments only under matched processing and scoring conditions (standard IHC practice).

Can this stain distinguish CD1B isoforms or reveal epitope masking?

CD1B has 2 reported isoforms, but the supplied antibody caption does not identify its epitope or establish isoform selectivity (UniProt P29016 isoforms; datasheet A02158-1). The precursor has a signal sequence at residues 1–17, and the mature chain begins at residue 18 (UniProt P29016 processing). Reported glycosylation sites are 38, 75, 146, and 258; their effect on this antibody’s staining is unknown (UniProt P29016 glycosylation; datasheet A02158-1). Interpret a negative paraffin-section stain only after checking retrieval, positive-control tissue, and detection, and do not assign an isoform from chromogenic intensity alone (standard IHC practice).

How would I investigate CD1B localisation with a complementary IF experiment?

Use IF as a separate localisation check while keeping chromogenic paraffin-section IHC as this antibody’s documented tissue workflow (datasheet A02158-1). For cortical thymocytes, multiplex CD1B with a T-cell marker such as CD3 and assess cell identity alongside membrane and intracellular signal (UniProt P29016 tissue specificity; standard IF practice). Choose a far-red fluorophore when tissue autofluorescence obscures shorter wavelengths, and include single-label and no-primary controls (standard IF practice). If the antibody recognises an extracellular epitope, compare staining before and after permeabilisation; access to the 325–333 cytoplasmic tail or intracellular pools requires permeabilisation (UniProt P29016 topology; standard IF practice). The epitope’s location is unreported for this antibody (datasheet A02158-1).

How can I separate CD1B signal from DAB background?

The documented paraffin-section workflow used 10% goat serum block, biotinylated secondary antibody, streptavidin-biotin detection, and DAB (datasheet A02158-1). Check endogenous peroxidase and endogenous biotin background with appropriate control sections, since either can produce chromogen independent of primary-antibody binding in this workflow (standard IHC practice). Include a no-primary control and compare it with tissue stained at the documented 1 μg/ml primary concentration (datasheet A02158-1; standard IHC practice). If diffuse staining persists, assess washing, blocking, section edges, and chromogen development time while preserving the same settings across controls and specimens (standard IHC practice).

How should I quantify CD1B-positive cells in chromogenic sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the scored cell population and compartment before measuring CD1B, because both surface and endosomal or lysosomal localisation are reported (UniProt P29016 localisation). For a specified cell population, record the percentage of positive viable cells and an intensity-based H-score from 0–300 using fixed thresholds (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of viable tissue and state whether membrane, punctate cytoplasmic, or combined signal was counted (standard IHC practice). Normalise counts to the eligible cells or viable tissue area actually examined, excluding necrosis and section artefacts under the same rules for every specimen (standard IHC practice).

What makes a CD1B-positive IHC pattern convincing?

A convincing pattern has staining in intact cells with interpretable membrane or intracellular punctate localisation, consistent with reported CD1B trafficking (UniProt P29016 localisation). Evaluate cell identity cautiously: lymphoid-tissue RNA enrichment and cortical-thymocyte expression provide context, but HPA reports low consistency between tissue antibody staining and RNA data (HPA tissue IHC; UniProt P29016 tissue specificity). Compare the pattern with no-primary and positive-control sections, and reject edge staining, necrotic deposits, or endogenous peroxidase signal as evidence of CD1B (standard IHC practice). The catalog tonsil image establishes a paraffin-section staining example, but its caption does not identify the stained cell type or fixative (datasheet A02158-1).

Boster reagents

Best CD1B / T-cell surface glycoprotein CD1b IHC Antibodies

The IHC-validated catalog antibody has a chromogenic IHC figure from human paraffin-embedded tonsil; no IF/ICC figure or non-human reactivity is listed (A02158-1 image caption; catalog applications and reactivity).

Real IHC data

Anti-CD1b Antibody ®

Cat # A02158-1

View product →

A02158-1 is listed for human IHC and WB, with an IHC figure from a paraffin-embedded human tonsil section (catalog applications and reactivity; A02158-1 image caption). The caption reports citrate retrieval at pH 6 and DAB detection; it does not report the fixative (A02158-1 image caption).

Which to pick: For tissue IHC, choose A02158-1, a rabbit polyclonal antibody listed for human IHC and illustrated on a paraffin-embedded human tonsil section (catalog host, clonality, applications and reactivity; A02158-1 image caption). No IF/ICC-validated SKU is listed: A02158-1 lists IHC and WB and has no IF figure (catalog applications and IF images). No SKU is listed for cross-species work; A02158-1 lists human reactivity only (catalog reactivity).

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.