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IL2RB / Interleukin-2 receptor subunit beta · IHC design guide
Design Immunohistochemistry for IL2RB
Plan paraffin-section IL2RB staining around selective cytoplasmic signal in immune cells (HPA tissue IHC). The catalog antibody’s IHC example uses 2 μg/mL primary antibody with chromogenic detection (datasheet A03401-3).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IL2RB 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 staining in a subset of immune cells (HPA tissue IHC) | |
| Staining pattern | Selective cytoplasmic signal in some immune cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03401-3) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Adipose tissue+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 | Sparse positive cells may be missed during scoring (HPA tissue IHC) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; extracellular versus cytoplasmic epitope matters (UniProt) |
Section 1
Recommended IL2RB IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A03401-3) with the published MSI-H colon cancer tissue microarray protocol (PMC8057496).
| Sample | Paraffin-embedded human appendicitis tissue; fixative not specified (datasheet A03401-3) |
| Fixation | Image fixative and duration unreported (datasheet A03401-3); 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 A03401-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03401-3) |
| Primary antibody | Rabbit anti-IL2RB, 2-5μg/ml (datasheet A03401-3) |
| Primary incubation | Overnight at 4 °C (datasheet A03401-3) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03401-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | IL2RB-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: Medium). HPA tissue profile: Selective cytoplasmic expression in a subset of immune cells. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A03401-3); the published protocol used ER2 for 20 min (PMC8057496).
Section 2
What Is the Expected IL2RB Staining Pattern?
IL2RB is a cell-membrane receptor with an extracellular segment at residues 27–240 and a cytoplasmic segment at 266–551 (UniProt P14784 topology). In tissue IHC, expect selective cytoplasmic staining in a subset of immune cells (HPA tissue IHC: Enhanced reliability). Interpret that observed IHC pattern alongside the membrane assignment; do not require a crisp membrane rim in every positive paraffin-section cell (HPA tissue IHC; UniProt P14784 subcellular location).
| Subset of immune cells stain at medium intensity in lymphoid tissue or bone marrow. | This fits the reported selective cytoplasmic IHC profile (HPA tissue IHC: Enhanced reliability). Medium staining is reported in bone-marrow hematopoietic cells and non-germinal-center cells of lymph node and tonsil (HPA tissue IHC: Medium). Judge the labelled cells, since the HPA profile describes a subset rather than uniform staining throughout those tissues (HPA tissue IHC). |
| Signal appears mainly nuclear, or concentrates in structures outside the expected immune-cell subset. | A nuclear pattern does not match the reported selective cytoplasmic tissue staining or cell-membrane assignment (HPA tissue IHC; UniProt P14784 subcellular location). Treat it as suspect until controls and cell identity are checked (standard IHC practice). HPA notes that some stained cells or structures are unannotated, so morphology alone may not identify every positive cell (HPA tissue IHC: reliability description). |
| Adipocytes or respiratory epithelial cells stain as strongly as the proposed positives. | Those cell types are reported as not detected in adipose tissue and bronchus, respectively (HPA tissue IHC: Not detected). Their staining raises possible cross-reactivity or endogenous detection activity; it does not by itself establish which cause applies (standard IHC practice). Compare the suspect signal with an omitted-primary control and inspect cell morphology before scoring it as IL2RB (standard IHC practice). |
| Diffuse colour covers cells, stroma, and empty areas without a discrete positive subset. | That distribution conflicts with selective staining of immune cells (HPA tissue IHC). General IHC causes include nonspecific antibody binding, incomplete blocking, or detection-reagent background (standard IHC practice). Check whether the same haze appears in the omitted-primary control, then assess the primary-antibody and detection steps separately (standard IHC practice). |
| No labelled cells appear in a bone-marrow or lymph-node section. | HPA reports medium staining in bone-marrow hematopoietic cells and lymph-node non-germinal-center cells (HPA tissue IHC: Medium). A blank section therefore warrants a control review, although positivity is confined to particular cells rather than every cell in either tissue (HPA tissue IHC). Confirm that relevant cells are present before adjusting retrieval or detection conditions (standard IHC practice). |
💡Expected IL2RB appearanceCall a result positive when a subset of immune cells shows selective, approximately medium cytoplasmic IHC staining (HPA tissue IHC: profile and positive levels); broad nuclear, stromal, or negative-cell staining is suspect rather than the expected pattern (HPA tissue IHC; standard IHC interpretation).
| Observed compartment versus receptor topology | HPA describes selective cytoplasmic staining in tissue IHC, while UniProt assigns IL2RB to the cell membrane (HPA tissue IHC; UniProt P14784 subcellular location). Use the reported IHC appearance to assess the slide without treating a missing sharp membrane rim as an automatic failure (HPA tissue IHC; standard IHC interpretation). |
| Cell and tissue selection | HPA reports medium signal in bone-marrow hematopoietic cells, endometrial lymphocytes, splenic red-pulp cells, and non-germinal-center cells in lymph node and tonsil (HPA tissue IHC: Medium). HPA reports several other specified cell populations as not detected; these are cell-level comparisons, not whole-tissue absence claims (HPA tissue IHC: Not detected). |
| Antibody evidence | The listed antibody HPA062657 has Enhanced IHC validation, and the tissue profile has Enhanced reliability based on staining and RNA-expression consistency (HPA antibodies: HPA062657; HPA tissue IHC: reliability description). This supports the reported pattern but does not establish that every stained structure has been identified (HPA tissue IHC: reliability description). |
| Epitope and processing limits | UniProt places the mature chain at residues 27–551 and lists four glycosylation sites in its extracellular region (UniProt P14784 processing, topology, glycosylation). The supplied record does not locate the IHC antibody epitope, so these annotations alone cannot predict how a particular retrieval condition will change staining (UniProt P14784; standard IHC interpretation). |
| IF/ICC: what can this record establish? | HPA summarizes IL2RB as membrane-localized for ICC-IF but supplies no main-location assignment or cell-line images here (HPA subcellular ICC-IF). That summary is consistent with the UniProt membrane assignment; it does not establish an IF/ICC staining pattern for a particular cell line or override the observed tissue-IHC profile (HPA subcellular ICC-IF; UniProt P14784). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive tissue is entirely blank. | The relevant positive-cell subset may be absent from the section, or the IHC workflow may have failed (HPA tissue IHC: selective profile; standard IHC practice). | First verify hematopoietic cells in bone marrow or non-germinal-center cells in lymph node; both have reported medium staining (HPA tissue IHC: Medium). Then check the run control, primary-antibody step, retrieval, and chromogenic detection using the validated IHC procedure (standard IHC practice). |
| Only a few cells stain, making the section look mostly negative. | Sparse signal can reflect the reported selective immune-cell pattern rather than assay failure (HPA tissue IHC: profile). | Score the specific cell population and its compartment instead of averaging across the section (HPA tissue IHC: selective cytoplasmic profile; standard IHC interpretation). Compare a second reported positive cell population if the candidate cells are hard to identify (HPA tissue IHC: positive list). |
| Adipocytes or bronchial respiratory epithelium stain. | These cells are reported as not detected; cross-reactivity or endogenous chromogenic activity is possible (HPA tissue IHC: Not detected; standard IHC practice). | Compare an omitted-primary control, check the relevant endogenous-activity block, and reassess primary-antibody specificity before calling the signal IL2RB (standard IHC practice). Keep the comparison at cell level rather than labelling the entire tissue negative (HPA tissue IHC). |
| Background is diffuse across the section. | Broad staining departs from HPA's selective immune-cell profile and may arise from nonspecific binding or detection background (HPA tissue IHC; standard IHC practice). | Inspect the omitted-primary control, confirm blocking and washes, and adjust the primary-antibody or detection conditions only after locating the background source (standard IHC practice). Recheck whether a discrete immune-cell subset remains visible (HPA tissue IHC: profile). |
| Staining is predominantly nuclear. | A nuclear-dominant result disagrees with the reported cytoplasmic IHC profile and membrane assignment (HPA tissue IHC; UniProt P14784 subcellular location). | Review counterstain and chromogen separation, compare controls, and check whether the nuclear colour tracks with the primary antibody before interpreting it as target signal (standard IHC practice). |
| Changing retrieval seems to alter the signal. | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Use the catalog antibody's validated IHC-P procedure as the starting condition, then compare conditions with the same reported positive-cell population and appropriate run controls (standard IHC practice; HPA tissue IHC: positive list). Avoid inferring an IL2RB-specific fixation mechanism from signal changes alone (standard IHC interpretation). |
Experiment evidence
IHC & IF images showing IL2RB staining
A03401-3 · IHCIHC analysis of IL2 Receptor beta/p75/IL2RB using anti-IL2 Receptor beta/p75/IL2RB antibody (A03401-3). IL2 Receptor beta/p75/IL2RB was dete
A03401-3 · IFIF analysis of IL2 Receptor beta/p75/IL2RB using anti- IL2 Receptor beta/p75/IL2RB antibody (A03401-3). IL2 Receptor beta/p75/IL2RB was dete
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100385-IL2RB/tissue/cerebral+cortex#imid_17214379
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100385-IL2RB/tissue/colon#imid_17214305
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100385-IL2RB/tissue/liver#imid_17214290
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000100385-IL2RB/tissue/kidney#imid_17214299
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IL2RB IHC & IF
🧪Run lymph node first and look for staining in non-germinal center cells, where IL2RB is reported at a Medium level (HPA: lymph node, non-germinal center cells, Medium). Use adrenal gland as the negative tissue and assess its glandular cells, where IL2RB is Not detected (HPA: adrenal gland, glandular cells, Not detected). On the lymph node slide, use nonimmune stromal cells as provisional internal negatives: they should lack specific membrane staining, but their negative status is not established by the supplied HPA row (HPA: lymph node, non-germinal center cells, Medium; UniProt P14784: cell membrane).
Positive control tissue: Bone marrow (Hematopoietic cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for IL2RB; derive a cell-line control from the positive tissue's cell type (Hematopoietic cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG isotype control, and IL2RB-knockout material if available as a biological negative (selected-SKU caption: rabbit primary antibody; standard IHC practice). In lymph node, quench endogenous peroxidase and control for endogenous biotin because the reported detection uses a biotinylated secondary and streptavidin-biotin complex with DAB (selected-SKU caption: biotinylated goat anti-rabbit IgG, SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). The reported IHC result used heat retrieval in EDTA at pH 8.0, but the evidence does not establish whether retrieval is required for every specimen (selected-SKU caption: heat-mediated EDTA retrieval). No matched evidence establishes that frozen sections or IF are easier for IL2RB; for chromogenic lymph-node IHC, assess background from endogenous peroxidase and biotin alongside the membrane staining pattern (UniProt P14784: cell membrane; selected-SKU caption: SABC/DAB; standard IHC practice).
HPA tissue IHC evidence for IL2RB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Endometrium | Lymphocytes | Medium | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Medium | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | Medium | 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 → |
| Appendix | Glandular 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 IL2RB IHC Tips
Use the catalog antibody’s paraffin-section evidence and IL2RB’s membrane topology to guide staining, controls, and interpretation (caption A03401-3; UniProt P14784 topology).
Which retrieval conditions should I start with for IL2RB in paraffin sections?
Start with heat-mediated EDTA at pH 8.0 for paraffin-section IL2RB IHC (datasheet A03401-3). The selected human appendicitis section received this retrieval, followed by 10% goat-serum blocking and 2 μg/ml primary antibody overnight at 4°C (caption A03401-3). If staining is weak, vary heating duration or test another retrieval buffer only as a controlled fallback, processing adjacent sections together and recording tissue morphology (standard IHC practice). Assess immune-cell-rich areas alongside glandular cells: HPA reports selective staining in immune cells and no detection in appendix glandular cells (HPA tissue IHC).
How should I handle fixation when IL2RB staining varies between specimens?
The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown (caption A03401-3). Record each specimen’s fixative, time in fixative, processing history, and section age before comparing staining intensity (standard IHC practice). Keep section thickness, EDTA retrieval at pH 8.0, primary concentration, and development time matched across a comparison series (datasheet A03401-3; standard IHC practice). If a specimen stains weakly, compare adjacent sections under controlled retrieval conditions and inspect morphology before attributing the difference to fixation; include a staining control with immune-cell-rich tissue from the same run (standard IHC practice; HPA tissue IHC).
Should IL2RB staining appear at the membrane or in the cytoplasm?
Expect a membrane-associated signal because IL2RB has an extracellular region at residues 27–240, a transmembrane segment at 241–265, and a cytoplasmic region at 266–551 (UniProt P14784 topology). HPA also describes selective cytoplasmic staining in a subset of immune cells, so compartment calls need to be tied to the antibody and specimen examined (HPA tissue IHC). IL2RB participates in receptor-mediated endocytosis, which makes intracellular signal biologically plausible but does not establish the identity of every cytoplasmic deposit (UniProt P14784 function). Compare membrane and cytoplasmic patterns in morphologically intact immune cells, and treat diffuse nuclear or extracellular DAB as suspect until controls support it (standard IHC practice; UniProt P14784 subcellular location).
How could epitope position affect staining after retrieval?
The supplied record lists 0 isoforms and places the extracellular, transmembrane, and cytoplasmic regions at residues 27–240, 241–265, and 266–551, respectively (UniProt P14784). The selected caption does not identify the catalog antibody’s epitope, so its membrane-facing side cannot be assigned from that image (caption A03401-3). Extracellular residues 29, 43, 71, and 149 are annotated glycosylation sites; epitope accessibility should therefore be assessed with the actual antibody rather than inferred from sequence alone (UniProt P14784 glycosylation; standard IHC practice). When comparing antibodies, document each mapped epitope and hold retrieval, detection, and scoring conditions constant across adjacent sections (standard IHC practice).
How can I assess IL2RB by IF alongside immune-cell markers?
For exploratory IF, multiplex IL2RB with a marker that identifies the immune-cell population under study, since HPA reports selective expression in a subset of immune cells (HPA tissue IHC). Choose fluorophores after checking unstained tissue for autofluorescence, and place the weaker signal in a spectrally cleaner channel when possible (standard IF practice). Permeabilise for an intracellular epitope, but minimise permeabilisation when testing an accessible extracellular epitope; the catalog antibody’s epitope side is not specified in the selected caption (UniProt P14784 topology; caption A03401-3; standard IF practice). Include single-colour and no-primary controls, then compare cellular overlap and membrane versus intracellular distribution before interpreting multiplex signal (standard IF practice; UniProt P14784 subcellular location).
What should I check when DAB background obscures IL2RB-positive cells?
First compare a no-primary section with the stained section to separate detection-system deposits from antibody-dependent signal (standard IHC practice). The selected workflow uses a biotinylated secondary, streptavidin-biotin complex, and DAB, making endogenous biotin and peroxidase activity relevant background checks (caption A03401-3; standard IHC practice). Use an appropriate peroxidase block, assess endogenous biotin where needed, and control DAB development time on matched sections (standard IHC practice). If diffuse staining persists, reassess blocking, washes, and primary concentration against the caption’s 10% goat serum and 2 μg/ml primary conditions, while preserving tissue morphology (caption A03401-3; standard IHC practice).
How should I score heterogeneous IL2RB staining in tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HPA reports staining in a subset of immune cells rather than uniform tissue-wide expression (HPA tissue IHC). Record the percentage of positive eligible cells and staining intensity, or calculate an H-score from 0–300 with fixed intensity categories (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of analysed tissue and normalise counts to the number of eligible immune cells when their abundance differs between samples (standard IHC practice; HPA tissue IHC). Apply the same threshold, section selection, counterstain, and DAB development conditions across samples, and exclude damaged areas using predefined rules (standard IHC practice).
How can I distinguish true IL2RB staining from artefact?
Prioritise staining in morphologically intact immune cells: HPA reports selective immune-cell expression, including lymphocytes in endometrium, while appendix glandular cells were not detected (HPA tissue IHC). Membrane-associated staining fits IL2RB topology, and some intracellular staining is plausible given its endocytic role and HPA’s cytoplasmic observation (UniProt P14784 topology and function; HPA tissue IHC). Treat isolated nuclear signal, tissue-edge deposits, necrotic-area staining, or signal reproduced without primary antibody as possible artefact (standard IHC practice). Check a no-primary control and peroxidase-block performance before assigning DAB deposits to IL2RB, especially with the selected biotin-complex detection workflow (caption A03401-3; standard IHC practice).
Boster reagents
Best IL2RB / Interleukin-2 receptor subunit beta IHC Antibodies
A03401-3 has human paraffin-section IHC and A549-cell IF images (catalog image captions). The catalog lists human, mouse and rat reactivity (catalog: reactivity).
Real IHC data
Anti-IL2 Receptor beta/p75/IL2RB Antibody ®
Cat # A03401-3
A03401-3 was demonstrated by IHC on a paraffin-embedded human appendicitis section (A03401-3 IHC caption). The same SKU was demonstrated by IF in A549 cells and lists IHC, IF and ICC applications (A03401-3 IF caption; catalog: applications).
Which to pick: For tissue IHC, choose A03401-3: its own image shows a human paraffin section with EDTA pH 8.0 retrieval and 2 μg/ml primary antibody; the fixative is unreported (A03401-3 IHC caption). For IF/ICC, A03401-3 lists both applications and has an A549-cell IF image using 5 μg/ml primary antibody (catalog: applications; A03401-3 IF caption). For mouse or rat samples, A03401-3 lists reactivity to both species, although its supplied IHC and IF images show human samples; clonality is unreported (catalog: reactivity and clone; A03401-3 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P14784 (IL2RB_HUMAN, Interleukin-2 receptor subunit beta).
- Human Protein Atlas. IL2RB tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. IL2RB subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. IL2RB antibody validation summary (1 antibodies).
- Evolutionary genetic algorithm identifies IL2RB as a potential predictive biomarker for immune-checkpoint therapy in colorectal cancer. NAR genomics and bioinformatics 2021 — PMC8057496.
- Human interleukin-2 receptor β mutations associated with defects in immunity and peripheral tolerance. The Journal of experimental medicine 2019 — PMC6547869.
- PubMed PMID:2785715 — UniProt-cited evidence.
- PubMed PMID:15461802 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IL2RB (Interleukin-2 receptor subunit beta). Expected localisation Cytoplasmic staining in a subset of immune cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03401-3). Validated IHC antibody: A03401-3 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human appendicitis tissue; fixative not specified (datasheet A03401-3); 2. Fixation: Image fixative and duration unreported (datasheet A03401-3); 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: EDTA pH 8.0 (datasheet A03401-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03401-3); 8. Primary antibody: Rabbit anti-IL2RB, 2-5μg/ml (datasheet A03401-3); 9. Primary incubation: Overnight at 4 °C (datasheet A03401-3). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Sparse positive cells may be missed during scoring (HPA tissue IHC); Isoform / epitope: No isoforms annotated; extracellular versus cytoplasmic epitope matters (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt P14784, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A03401-3, PMC8057496. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IL2RB/immunohistochemistry-ihc-guide.