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IGF2BP2 / Insulin-like growth factor 2 mRNA-binding protein 2 · IHC design guide
Design Immunohistochemistry for IGF2BP2
Plan IGF2BP2 paraffin IHC around mainly cytoplasmic tissue staining (HPA tissue IHC). This guide covers fixation consistency, controls and scoring with an IHC-validated antibody at 1:50 (datasheet: M02010-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IGF2BP2 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 | Mainly cytoplasmic in tissue IHC (HPA tissue IHC) | |
| Staining pattern | Mainly cytoplasmic; high in several glandular tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M02010-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image M02010-1) | |
| Caveat | Staining and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | Cellular stress recruits IGF2BP2 to stress granules (UniProt) | |
| Isoform / epitope | 6 isoforms; check epitope coverage across variants (UniProt) |
Section 1
Recommended IGF2BP2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: M02010-1) is accompanied by four published IGF2BP2 chromogenic IHC protocols (PMC10227963; PMC12920223; PMC9923020; PMC13113562).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet M02010-1) |
| Fixation | Image fixative and duration unreported (datasheet M02010-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet M02010-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M02010-1) |
| Primary antibody | Rabbit monoclonal (clone 24I46) anti-IGF2BP2, 1:50 (datasheet M02010-1) |
| Primary incubation | Overnight at 4 °C (datasheet M02010-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M02010-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | IGF2BP2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Mainly cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: M02010-1). Citrate at pH 6.0 is a published comparison condition (PMC12920223).
Section 2
What Is the Expected IGF2BP2 Staining Pattern?
IGF2BP2 should appear mainly in the cytoplasm of stained cells on paraffin sections (HPA: mainly cytoplasmic expression in most tissues). High staining is reported in selected glandular cells, hematopoietic cells, cardiomyocytes and myocytes (HPA: tissue IHC). Nuclear localisation is also possible (UniProt Q9Y6M1: subcellular location), while membrane staining lacks support from its topology (UniProt Q9Y6M1: no transmembrane segment). HPA rates tissue IHC reliability Enhanced, with medium consistency between staining and RNA data (HPA: tissue IHC).
| Cytoplasmic staining in adrenal, duodenal, gallbladder, parathyroid or stomach glandular cells. | This matches the reported compartment and high-staining cell types (HPA: mainly cytoplasmic; High in those glandular cells). Judge the named cells against adjacent tissue rather than treating all cells in the section as equally positive (general IHC interpretation). |
| Discrete cytoplasmic puncta, or some nuclear signal alongside cytoplasmic staining. | Puncta can fit localisation to cytoplasmic mRNP granules, P-bodies or stress granules; nuclear localisation is also annotated (UniProt Q9Y6M1: subcellular location). A punctate pattern alone does not identify a granule type (general IF/IHC interpretation). |
| Predominantly cell-surface or extracellular staining with little cytoplasmic signal. | That pattern conflicts with the reported mainly cytoplasmic tissue pattern and absence of a transmembrane segment (HPA: tissue IHC; UniProt Q9Y6M1: topology). Treat it as suspect and compare it with a validated positive section and controls (general IHC practice). |
| Strong colour in unexpected cells, especially where staining follows tissue edges or pigment. | The distribution may reflect cross-reactivity or endogenous detection activity (general chromogenic IHC practice). HPA lists high staining for particular cell types, not every cell in those tissues (HPA: tissue IHC); confirm cell identity and inspect a primary-omission control. |
| Haze across cells and empty tissue spaces, or no signal in a reported high-staining section. | Widespread haze limits localisation and may reflect nonspecific detection; absent signal may reflect a workflow failure (general IHC practice). Neither finding alone establishes IGF2BP2 absence, since HPA reports high staining in selected cells of those tissues (HPA: tissue IHC). |
💡Expected IGF2BP2 appearanceCall a section positive when the expected cell population shows discernible, mainly cytoplasmic staining, potentially strong in HPA high-staining populations; diffuse tissue-wide haze or isolated surface staining is suspect (HPA: tissue IHC; UniProt Q9Y6M1: topology; general IHC interpretation).
| Choice of positive section | Use a section containing one of the reported High populations, such as adrenal glandular cells or heart cardiomyocytes (HPA: tissue IHC). High describes staining in the specified cells, so inspect those cells when assessing a run (HPA: tissue IHC; general IHC interpretation). |
| Low-staining comparison | HPA reports Low staining in cerebellar granular-layer cells, hippocampal and caudate neurons, bladder urothelium, prostate glandular cells and ovarian stroma (HPA: tissue IHC). Low is not a documented negative control; weak staining there does not by itself invalidate the assay. |
| Compartment and molecular form | IGF2BP2 is a 599-residue chain with six annotated isoforms and no signal peptide, propeptide or transmembrane segment (UniProt Q9Y6M1: processing, isoforms, topology). These annotations support an intracellular readout; they do not identify which isoform an antibody detects. |
| Evidence strength | HPA labels tissue IHC Enhanced but describes only medium consistency with RNA expression (HPA: tissue IHC). HPA035145 and CAB017126 each have Enhanced IHC and Supported ICC status (HPA: antibody validation); these ratings do not validate an unspecified catalog antibody. |
| Q: What should IF/ICC show? | A: Cytosolic staining is the HPA ICC-IF result (HPA: subcellular, cytosol enhanced). UniProt also annotates nucleus and cytoplasmic granules (UniProt Q9Y6M1: subcellular location). Interpret any additional compartment with controls; IF/ICC conditions belong in its own guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported high-staining cells are blank. | A failed detection step, unsuitable antibody conditions or absent target in the sampled cells are possibilities (general IHC practice); HPA's High designation does not guarantee every section is positive (HPA: tissue IHC). | Confirm the expected cell population is present, check a concurrently stained positive section, then review antibody dilution, retrieval and detection against the chosen assay instructions (general IHC practice). |
| Colour covers tissue spaces and many unrelated structures. | Nonspecific binding or incomplete blocking or washing may produce diffuse background (general chromogenic IHC practice); this obscures the mainly cytoplasmic pattern expected from HPA (HPA: tissue IHC). | Compare a primary-omission control; review blocking, wash stringency, antibody dilution and detection development time (general IHC practice). Score only cell-associated staining that remains interpretable. |
| Staining is concentrated at the surface of cells. | A membrane-dominant result is discordant with the reported cytoplasmic pattern and lack of a transmembrane segment (HPA: tissue IHC; UniProt Q9Y6M1: topology). Nonspecific staining is one possibility (general IHC interpretation). | Inspect a reported high-staining cell population and the primary-omission control; review antibody specificity and detection background before assigning a compartment (general IHC practice). |
| Nuclear staining dominates with little cytoplasmic staining. | Nuclear localisation is annotated, but HPA describes mainly cytoplasmic tissue staining (UniProt Q9Y6M1: subcellular location; HPA: tissue IHC). Nuclear dominance alone cannot establish either specificity or artefact. | Compare matched positive tissue and omission controls; record nuclear and cytoplasmic staining separately before interpreting the deviation (general IHC interpretation). |
| An apparently low-staining comparison shows weak positive cells. | HPA's Low category denotes observed staining, not verified absence (HPA: tissue IHC). Weak signal may therefore be compatible with the reference pattern rather than a failed negative control. | Do not score the tissue as a zero-expression control; compare the named cell population and intensity with a reported High population in the same run (HPA: tissue IHC; general IHC interpretation). |
| Unexpected cells stain strongly while expected cells do not. | Cell misidentification, cross-reactivity or endogenous detection activity can distort the apparent distribution (general chromogenic IHC practice). HPA's High calls apply to specified cell types (HPA: tissue IHC). | Recheck morphology and the counterstain, inspect omission and detection controls, and repeat with an appropriately validated antibody if the discordance persists (general IHC practice; HPA: antibody validation). |
Experiment evidence
IHC & IF images showing IGF2BP2 staining
M02010-1 · IHCIHC analysis of IMP2 using anti-IMP2 antibody (M02010-1). IMP2 was detected in a paraffin-embedded section of human lung cancer tissue. Heat
M02010-1 · IHCIHC analysis of IMP2 using anti-IMP2 antibody (M02010-1). IMP2 was detected in a paraffin-embedded section of human testicular germ cell tum
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000073792-IGF2BP2/tissue/cerebral+cortex#imid_14059986
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000073792-IGF2BP2/tissue/colon#imid_14059920
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000073792-IGF2BP2/tissue/liver#imid_14059921
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000073792-IGF2BP2/tissue/kidney#imid_14059906
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017126; green = IGF2BP2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017126; green = IGF2BP2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IGF2BP2 IHC & IF
🧪Run bone marrow first; hematopoietic cells should stain (HPA: High in bone marrow hematopoietic cells). HPA detects IGF2BP2 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat any unstained cells on the positive slide as a background reference only after confirming their identity and staining pattern (HPA: no negative tissue rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: IGF2BP2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show IGF2BP2 in A-431, U-251MG, U2OS, ASC52telo, with annotated localisation: Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section, an isotype control matched to the primary antibody’s host species and class, and an IGF2BP2 knockout biological control where available (standard IHC practice; caption: rabbit primary antibody). Quench endogenous peroxidase and check for residual DAB signal in bone marrow, where myeloid cells can have endogenous peroxidase activity (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected M02010-1 paraffin-section caption does not state the fixative (selected-SKU tissue-IHC caption). That caption uses heat-mediated EDTA retrieval at pH 8.0, but does not establish that retrieval is required; the supplied evidence also does not establish that frozen sections or IF are easier (selected-SKU tissue-IHC caption; HPA subcellular). In bone marrow, endogenous myeloid peroxidase can complicate chromogenic interpretation, so compare staining with the peroxidase-quenched no-primary control (standard IHC practice).
HPA tissue IHC evidence for IGF2BP2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: IGF2BP2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced IGF2BP2 IHC Tips
Troubleshoot IGF2BP2 staining in paraffin sections by checking retrieval, controls, cellular localisation and scoring before interpreting chromogenic signal.
Which retrieval conditions should I start with for IGF2BP2 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet M02010-1). That condition was used for the catalog antibody in a paraffin-embedded human lung cancer section; the caption does not report retrieval time or section fixative (datasheet M02010-1). Keep heating and cooling conditions consistent across comparison sections, then check whether cytoplasmic staining remains distinct from background (standard IHC practice; HPA tissue IHC). If staining is weak, optimise heating time on matched sections before testing another retrieval buffer or pH, with the same positive and no-primary controls (standard IHC practice).
How should I troubleshoot weak IGF2BP2 staining when fixation history is uncertain?
Record the fixative and fixation duration for each specimen before comparing IGF2BP2 staining, because target-specific fixation sensitivity is unknown from the supplied evidence (datasheet M02010-1; standard IHC practice). The selected image documents a paraffin section but does not state its fixative, so it cannot establish a fixation condition for this antibody (datasheet M02010-1). Compare sections with known processing histories using the same EDTA pH 8.0 retrieval and detection run (datasheet M02010-1; standard IHC practice). Include a consistently processed positive control and a no-primary control to distinguish processing variation from general chromogenic background (standard IHC practice).
How should I assess nuclear or punctate staining alongside cytoplasmic IGF2BP2?
Expect mainly cytoplasmic staining across tissues when assessing this chromogenic assay (HPA tissue IHC). IGF2BP2 is also annotated in the nucleus, P-bodies and stress granules, with stress-granule recruitment reported under cellular stress (UniProt Q9Y6M1). Record diffuse cytoplasmic, nuclear and punctate signal separately rather than treating every stained compartment as equivalent (UniProt Q9Y6M1; standard IHC practice). Check whether puncta recur in intact cells away from damaged edges, and compare them with no-primary background on an adjacent section (standard IHC practice). A stress-granule interpretation needs a suitable marker and experimental context; morphology alone does not identify the structure (UniProt Q9Y6M1; standard IHC practice).
Could isoforms or epitope accessibility explain different staining between specimens?
IGF2BP2 has 6 listed isoforms, but the supplied antibody caption does not map the recognised epitope or establish isoform coverage (UniProt Q9Y6M1; datasheet M02010-1). Its RNA recognition motifs occupy residues 3–76 and 82–157, while four KH domains extend across the remaining annotated regions (UniProt Q9Y6M1). Annotated phosphorylation sites include serines 11, 162 and 164, plus threonine 550; their effects on this antibody's staining are unknown (UniProt Q9Y6M1; datasheet M02010-1). Compare matched specimens under identical retrieval and detection conditions, then seek epitope mapping or independent validation before attributing a staining difference to an isoform or modification (standard IHC practice).
How can IF clarify an ambiguous chromogenic IGF2BP2 pattern?
Use IF as a separate validation experiment when cell identity or compartment is unclear in the paraffin-section IHC result (standard IHC/IF practice). Multiplex IGF2BP2 with a marker for the expected cell type, and choose fluorophores whose emission is distinct from the specimen's measured autofluorescence (standard IF practice). Because IGF2BP2 has no transmembrane segment and is annotated in cytoplasmic and nuclear compartments, use permeabilisation appropriate for intracellular epitopes while checking that morphology remains intact (UniProt Q9Y6M1; standard IF practice). The supplied catalog image supports chromogenic tissue IHC, so establish IF antibody performance with appropriate single-stain and no-primary controls before comparing patterns across applications (datasheet M02010-1; standard IF practice).
What should I change when IGF2BP2 DAB staining is widespread or diffuse?
First inspect a no-primary section for nonspecific secondary or detection signal, and check the peroxidase block before DAB development (standard IHC practice). The catalog image used 10% goat serum blocking, a 1:50 primary incubation overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet M02010-1). If background persists, compare primary dilution and incubation changes on matched sections while holding retrieval and DAB development constant (standard IHC practice). Score cellular staining only after examining tissue edges, damaged areas and blank spaces for deposit that could mimic the mainly cytoplasmic pattern (HPA tissue IHC; standard IHC practice).
How should I quantify IGF2BP2 across paraffin sections with different cellularity? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring compartment before reading slides, because HPA reports mainly cytoplasmic tissue staining (HPA tissue IHC; standard IHC practice). For an intensity-weighted result, use an H-score from the percentages of cells at each staining intensity; also report the percentage of positive cells when that measure answers the study question (standard IHC practice). If spatial abundance matters, report positive-cell density per mm² of viable tissue and normalise counts to the measured area or relevant cell population (standard IHC practice). Apply one threshold across the batch, exclude necrosis and section edges by predefined rules, and retain the same positive-control section across runs (standard IHC practice).
What evidence separates convincing IGF2BP2 staining from tissue artefact?
A convincing result shows reproducible cellular signal in an expected compartment, with mainly cytoplasmic tissue staining as the reference pattern (HPA tissue IHC; standard IHC practice). High staining has been reported in adrenal glandular cells and bone-marrow hematopoietic cells, while low staining has been reported in prostate glandular cells; compare the correct cell population rather than whole-section darkness (HPA tissue IHC). Scrutinise staining confined to cut edges, necrotic areas or no-primary sections, and check endogenous peroxidase before interpreting DAB deposits (standard IHC practice). Nuclear signal remains biologically possible for IGF2BP2, but an unexpected dominant nuclear pattern needs independent compartment validation and compatible controls (UniProt Q9Y6M1; standard IHC practice).
Boster reagents
Best IGF2BP2 / Insulin-like growth factor 2 mRNA-binding protein 2 IHC Antibodies
Catalog antibodies list IHC reactivity in human, mouse, and rat (catalog applications and reactivity). IHC images show human paraffin sections; an IF image shows U2OS cells (catalog image captions).
Real IHC data
Anti-IMP2 Rabbit Monoclonal Antibody
Cat # M02010-1
The rendered M02010-1 card shows IHC in a paraffin-embedded human lung cancer section (M02010-1 IHC caption). A second caption documents a paraffin-embedded human testicular germ cell tumor section; the catalog lists IHC and IF/ICC applications with human, mouse, and rat reactivity (M02010-1 IHC caption; catalog applications and reactivity).
Which to pick: For tissue IHC, choose monoclonal M02010-1: its own caption documents EDTA pH 8.0 retrieval and 1:50 primary antibody on a human paraffin section; the fixative is unreported (M02010-1 IHC caption; catalog clone 24I46). For IF/ICC, choose A02010-4 because its IF caption documents U2OS cells stained with 1:100 primary antibody, fluorescent secondary antibody, and DAPI (A02010-4 IF caption). For mouse or rat samples, A02010-4, M02010-1, and M02010-2 all list reactivity, but the supplied IHC images document human tissue only; M02010-2 has no supplied IHC image (catalog reactivity and IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y6M1 (IF2B2_HUMAN, Insulin-like growth factor 2 mRNA-binding protein 2).
- Human Protein Atlas. IGF2BP2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. IGF2BP2 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. IGF2BP2 antibody validation summary (2 antibodies).
- A combined computational and functional approach identifies IGF2BP2 as a driver of chemoresistance in a wide array of pre-clinical models of colorectal cancer. Molecular cancer 2023 — PMC10227963.
- Predictive value of IGF2BP2 for endometrial cancer recurrence: a multicenter study. Frontiers in oncology 2026 — PMC12920223.
- Expression and significance of m6A-RNA-methylation in oral cancer and precancerous lesion. Frontiers in oncology 2023 — PMC9923020.
- IGF2BP2 Overexpression Predicts Poor Prognosis and Correlates with PD-L1 Expression in Intrahepatic Cholangiocarcinoma. Biomedicines 2026 — PMC13113562.
- PubMed PMID:10190901 — UniProt-cited evidence.
- PubMed PMID:16641997 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IGF2BP2 (Insulin-like growth factor 2 mRNA-binding protein 2). Expected localisation Mainly cytoplasmic in tissue IHC (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M02010-1). Validated IHC antibody: M02010-1 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet M02010-1); 2. Fixation: Image fixative and duration unreported (datasheet M02010-1); 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: EDTA pH 8.0 (datasheet M02010-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M02010-1); 8. Primary antibody: Rabbit monoclonal (clone 24I46) anti-IGF2BP2, 1:50 (datasheet M02010-1); 9. Primary incubation: Overnight at 4 °C (datasheet M02010-1). Watch for: Fixation: Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image M02010-1); Caveat: Staining and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: 6 isoforms; check epitope coverage across variants (UniProt); Regulation: Cellular stress recruits IGF2BP2 to stress granules (UniProt). Evidence: UniProt Q9Y6M1, HPA tissue IHC · Enhanced, HPA ICC-IF · cytosol., Datasheet M02010-1, PMC10227963, PMC12920223. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IGF2BP2/immunohistochemistry-ihc-guide.