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IGF2BP1 / Insulin-like growth factor 2 mRNA-binding protein 1 · IHC design guide
Design Immunohistochemistry for IGF2BP1
Plan IGF2BP1 IHC-P around cytoplasmic staining in testis duct and ovarian follicle cells (HPA tissue IHC). Use preleptotene spermatocytes in testis as a positive reference; their staining is reported as high (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IGF2BP1 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 testis and ovary (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in testis duct and ovarian follicle cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A02007-1) | |
| Positive control | Testis+2 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Use formaldehyde-fixed paraffin sections (selected-SKU IHC image A02007-1); keep fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Signal varies by tissue and cell type (HPA tissue IHC) | |
| Regulation | Embryonic; expressed in some cancers (UniProt) | |
| Isoform / epitope | 2 isoforms; verify C-terminal epitope coverage (UniProt; datasheet A02007-1) |
Section 1
Recommended IGF2BP1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published IGF2BP1 staining protocols for leiomyoma (PMC9154092), lung adenocarcinoma (PMC12967950), and osteosarcoma (PMC6449306).
| Sample | Formaldehyde-fixed, paraffin-embedded human testis tissue (datasheet A02007-1) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A02007-1); 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 (datasheet A02007-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-IGF2BP1, 1:25 (datasheet A02007-1) |
| Primary incubation | 1 hours at 37°C (datasheet A02007-1) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | IGF2BP1-positive staining in preleptotene spermatocytes of testis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in cells in seminiferous ducts of testis and follicle cells in ovary. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6 heat retrieval for the catalog antibody (datasheet A02007-1). The leiomyoma protocol instead reports CC1 conditioning (PMC9154092).
Section 2
What Is the Expected IGF2BP1 Staining Pattern?
IGF2BP1 is mainly cytoplasmic in tissue IHC, especially in testicular preleptotene spermatocytes and ovarian follicle cells; kidney collecting ducts can also stain (HPA: tissue IHC). Nuclear foci and peripheral RNA granules are possible localizations, although HPA ICC-IF supports mainly cytosolic signal (UniProt Q9NZI8: subcellular location; HPA: subcellular). It has no transmembrane segment (UniProt Q9NZI8: topology). HPA rates the tissue IHC pattern Enhanced, with medium consistency against RNA expression (HPA: tissue IHC).
| Strong cytoplasmic signal in preleptotene spermatocytes; moderate signal in follicle cells or collecting ducts. | This matches the reported cell types and relative levels: testis High, ovary Medium, kidney Medium (HPA: tissue IHC). Compare stained cells with the surrounding unstained cells, rather than treating the whole section as uniformly positive (standard IHC interpretation). |
| Predominantly nuclear staining, with little cytoplasmic signal, across many cells. | Reassess specificity and scoring: HPA describes mainly cytoplasmic tissue staining and supported cytosolic ICC-IF staining (HPA: tissue IHC; HPA: subcellular). Discrete nuclear foci remain biologically plausible, so a limited nuclear component alone is not proof of artefact (UniProt Q9NZI8: subcellular location). |
| Prominent signal in adipocytes or other cells reported as not detected. | Consider cross-reactivity or endogenous chromogen-generating activity before calling those cells IGF2BP1-positive (HPA: adipose tissue IHC; standard IHC interpretation). Check cell identity, staining boundaries and detection controls; HPA's negative finding applies to the specified cell population, not every cell in that tissue (HPA: tissue IHC). |
| Diffuse colour covers tissue, extracellular space or most cell types without clear cytoplasmic boundaries. | A widespread haze does not resemble HPA's cell-selective cytoplasmic pattern (HPA: tissue IHC). Background from detection reagents, incomplete blocking or excess antibody is possible (standard IHC practice); interpret the slide only after a matched negative detection control and tissue morphology are reviewed. |
| No visible staining in preleptotene spermatocytes on a testis section. | The expected High signal is missing (HPA: testis IHC). First verify that the relevant cells are present and identifiable; then check antibody validation, retrieval and detection controls (standard IHC practice). A blank slide alone cannot establish true absence of IGF2BP1. |
💡Expected IGF2BP1 appearanceCall the result positive when preleptotene spermatocytes show High, mainly cytoplasmic staining; broad nuclear-only staining or colour in reported negative cell types warrants control review (HPA: testis IHC; HPA: tissue IHC; UniProt Q9NZI8: subcellular location).
| Compartment and topology (UniProt Q9NZI8: topology; HPA: subcellular) | IGF2BP1 lacks a transmembrane segment, and supported ICC-IF localization is cytosolic (UniProt Q9NZI8: topology; HPA: subcellular). Score intracellular staining by compartment; do not treat membrane outlining as the expected target pattern (standard IHC interpretation). |
| Choice of comparison cells (HPA: tissue IHC) | Preleptotene spermatocytes provide a reported High reference; collecting ducts and follicle cells are Medium references (HPA: tissue IHC). Adipocytes and several listed cell populations are reported as not detected, making cell-level comparison useful within the stated tissue context (HPA: tissue IHC). |
| Evidence strength (HPA: tissue IHC; HPA: antibodies) | HPA calls the tissue result Enhanced but reports only medium agreement between antibody staining and RNA expression (HPA: tissue IHC). Two listed antibodies, HPA021367 and HPA062273, have Enhanced IHC validation (HPA: antibodies). These ratings support the reported pattern without validating every unexpected stained cell. |
| Isoforms and epitope coverage (UniProt Q9NZI8: isoforms) | UniProt lists 2 isoforms (UniProt Q9NZI8: isoforms). The supplied evidence gives no epitope location or isoform coverage for the IHC-validated antibodies; a staining difference cannot be assigned to an isoform from these records alone. |
| IF/ICC Q&A: what localization should be expected? (HPA: subcellular) | HPA reports mainly cytosolic ICC-IF signal, with images from CACO-2, Hep-G2 and U2OS; HPA021367 has Supported ICC validation (HPA: subcellular; HPA: antibodies). UniProt also lists nuclear foci and cytoplasmic granules (UniProt Q9NZI8: subcellular location). Use the separate IF/ICC guide for that application's procedure. |
| Situation | Likely cause | Next action |
|---|---|---|
| Testis positive-control cells appear blank. | The relevant preleptotene spermatocytes may be absent from the viewed area, or an IHC workflow step may have failed (HPA: testis IHC; standard IHC practice). | Locate the named cells on the counterstained section, then review the catalog antibody's IHC-P instructions and run a positive tissue and detection control (standard IHC practice). Do not infer a target-specific retrieval defect from HPA's staining level. |
| Colour is widespread in adipocytes or glandular cells reported as not detected. | Cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; standard IHC interpretation). The HPA negative calls refer to particular cell populations. | Compare morphology and cell identity with HPA's named populations; inspect an appropriate reagent-only detection control and endogenous-activity control for the chosen chemistry (standard IHC practice). |
| Nuclei dominate while the expected cytoplasmic pattern is weak. | The compartment pattern differs from HPA's mainly cytoplasmic tissue result, although UniProt allows discrete nuclear foci (HPA: tissue IHC; UniProt Q9NZI8: subcellular location). | Score nuclear and cytoplasmic compartments separately, compare with a known-positive section and review detection controls (standard IHC practice). Avoid calling every nuclear focus nonspecific solely from compartment. |
| A diffuse haze obscures cell boundaries. | Background from antibody concentration, blocking or chromogenic detection can obscure cell-selective staining (standard IHC practice). | Review the catalog antibody's IHC-P dilution guidance, blocking and reagent-only control; adjust the general workflow only after comparing control and test sections (standard IHC practice). No target-specific dilution is supplied here. |
| Ovary or kidney staining is weaker than testis. | That ordering is compatible with HPA's Medium follicle-cell and collecting-duct findings versus High preleptotene-spermatocyte staining (HPA: tissue IHC). | Compare the named cells, section morphology and controls before treating the lower intensity as failure (standard IHC interpretation). HPA's levels are qualitative and do not define a numeric scoring threshold. |
| IF/ICC shows cytosolic puncta or a small nuclear component. | UniProt places IGF2BP1 in cytoplasmic mRNP granules and describes discrete nuclear foci; HPA reports mainly cytosolic ICC-IF localization (UniProt Q9NZI8: subcellular location; HPA: subcellular). | Interpret those features against the supported cytosolic pattern and appropriate IF controls (HPA: subcellular; standard IF practice). The IHC tissue ratings do not themselves validate an IF finding (HPA: tissue IHC; HPA: antibodies). |
Experiment evidence
IHC & IF images showing IGF2BP1 staining
A02007-1 · IHCA02007-1 staining IGF2BP1 in human testis tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections
A02007-1 · IHCA02007-1 staining IGF2BP1 in human lung adenocarcinoma sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded se
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000159217-IGF2BP1/tissue/cerebral+cortex#imid_18540099
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000159217-IGF2BP1/tissue/colon#imid_18540017
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000159217-IGF2BP1/tissue/liver#imid_18540026
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000159217-IGF2BP1/tissue/kidney#imid_18540019
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA021367; green = IGF2BP1, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA021367; green = IGF2BP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IGF2BP1 IHC & IF
🧪Run testis first and expect staining in preleptotene spermatocytes (High; HPA: testis). Use adipose tissue as a negative tissue and expect no staining in adipocytes (HPA: adipocytes not detected); on the testis slide, cells outside the positive population should lack specific staining (HPA: preleptotene spermatocytes High).
Positive control tissue: Testis (Preleptotene spermatocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show IGF2BP1 in CACO-2, Hep-G2, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype-matched control using the primary antibody’s host species and clonality (standard IHC practice). A confirmed IGF2BP1 knockout sample or a validated peptide-block control can test target specificity (standard IHC practice). For chromogenic testis sections, check endogenous peroxidase and biotin background because the selected example uses a biotinylated secondary antibody (A02007-1 caption: biotinylated secondary).
⚠️Feasibility: The selected SKU’s fixative field is unreported (A02007-1 fixative field: not stated), and no target-specific fixation window or fixation effect is reported in the supplied evidence. The selected paraffin-section example uses heat retrieval in citrate buffer at pH 6 (A02007-1 caption); whether retrieval is essential for IGF2BP1 is unreported. The supplied evidence does not establish whether frozen sections or IF/ICC are easier; on testis sections, assess any biotin-associated background alongside the tissue signal (A02007-1 caption: biotinylated secondary).
HPA tissue IHC evidence for IGF2BP1
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 |
|---|---|---|---|---|
| Testis | Preleptotene spermatocytes | High | Protein (IHC) | HPA → |
| Kidney | Collecting ducts | Medium | Protein (IHC) | HPA → |
| Ovary | Follicle 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IGF2BP1 IHC Tips
Use matched controls and compartment-aware scoring to troubleshoot IGF2BP1 staining in paraffin sections; the immunofluorescence question addresses the separate IF/ICC workflow.
What should I change when IGF2BP1 staining is weak after retrieval?
Start with heat-mediated antigen retrieval in citrate buffer at pH 6 (datasheet A02007-1). Run a known positive testis section alongside the sample, and compare staining in seminiferous-duct cells rather than judging overall section color (HPA: cytoplasmic testis staining; standard IHC practice). The product image used a 1:25 primary dilution after this retrieval, so check antibody exposure and detection before changing buffer chemistry (caption A02007-1; standard IHC practice). If staining remains weak, test an alternative retrieval condition on adjacent sections, recording both signal and tissue damage; keep the citrate condition as the reference for comparison (standard IHC practice).
Could fixation explain weak or patchy IGF2BP1 staining?
Target-specific sensitivity of IGF2BP1 to fixation duration or fixative choice is unknown from the supplied evidence; assess fixation as a general IHC variable (standard IHC practice). Compare sections processed together and inspect whether weak staining tracks with tissue thickness, folds, or uneven processing (standard IHC practice). The catalog image documents paraffin sections and a 1:25 primary dilution, but it does not establish a fixation tolerance range for this antibody (caption A02007-1). If a controlled processing comparison is possible, hold retrieval and detection constant and score cytoplasmic signal in comparable cells across sections (HPA: cytoplasmic profile; standard IHC practice).
Should I accept nuclear or membrane-like staining as IGF2BP1?
IGF2BP1 is mainly cytosolic in the subcellular atlas, while UniProt also describes nuclear foci and cytoplasmic mRNP granules (HPA: supported cytosol; UniProt Q9NZI8 localisation). In paraffin IHC, score cytoplasmic staining separately from nuclear staining and compare each pattern with a known positive section (HPA: testis profile; standard IHC practice). A continuous cell-surface rim is less consistent with this protein because its record lists no transmembrane segment (UniProt Q9NZI8 topology). Treat an isolated nuclear or rim pattern as provisional until it survives retrieval and detection controls and recurs in the expected cell population (standard IHC practice; HPA: testis profile).
How should unknown epitope coverage affect IGF2BP1 IHC interpretation?
The record lists 2 isoforms and RNA-binding RRM and KH domains, but the supplied antibody evidence does not map its epitope (UniProt Q9NZI8 domains and isoforms; supplied catalog evidence). Do not assume that a negative section excludes both isoforms or that phosphorylation leaves antibody binding unchanged (UniProt Q9NZI8 isoforms and modified residues; standard IHC interpretation). If immunogen information becomes available, compare its sequence with each isoform and the listed modified residues before making an isoform claim (UniProt Q9NZI8 isoforms and modified residues; standard IHC practice). Meanwhile, report staining as antibody-detected IGF2BP1 signal and verify its cellular pattern with an independent assay where isoform identity matters (standard IHC practice).
How can I cross-check the IHC pattern with multiplex IF?
Use IF/ICC as a separate validation experiment and pair IGF2BP1 with an independently validated marker for the expected cell population, such as preleptotene spermatocytes in testis (HPA: High in preleptotene spermatocytes; standard IF practice). Select a fluorophore channel with low tissue autofluorescence and include single-color and no-primary controls before interpreting overlap (standard IF practice). Because IGF2BP1 lacks a transmembrane segment and is mainly cytosolic, permeabilise cells to reach intracellular epitopes, then check whether signal stays predominantly cytosolic (UniProt Q9NZI8 topology; HPA: supported cytosol; standard IF practice). Optimise IF fixation and permeabilisation empirically: the supplied paraffin IHC image does not establish IF/ICC fixation performance (caption A02007-1; standard IF practice).
What causes diffuse brown staining in my IGF2BP1 section?
First compare a no-primary control with the stained section to identify signal from secondary detection, endogenous enzyme activity, or pigment (standard chromogenic IHC practice). The catalog image used a biotinylated secondary and 3% BSA blocking for 0.5 hour; those conditions are a reference, not proof that every tissue is free of background (caption A02007-1). Apply a peroxidase block when using a peroxidase-based chromogen and assess endogenous biotin if using biotin-dependent detection (standard chromogenic IHC practice). A credible positive pattern should resolve in the expected cells and cytoplasm, while uniform stromal haze or staining in no-primary controls warrants detection troubleshooting (HPA: cytoplasmic testis profile; standard IHC practice).
How should I score heterogeneous IGF2BP1 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; in testis, preleptotene spermatocytes provide a documented positive population, and staining is chiefly cytoplasmic (HPA: High in preleptotene spermatocytes; HPA: tissue profile). Record the percentage of positive cells and intensity, or calculate an H-score from the same predefined population across slides (standard IHC practice). Normalise counts to the number of eligible cells, or report positive-cell density per mm² of viable tissue when cell counts are impractical (standard IHC practice). Use the same retrieval, antibody dilution, detection exposure, and scoring threshold for comparisons; exclude folds and necrotic areas before analysis (standard IHC practice).
When is an unexpected IGF2BP1-positive region likely artefactual?
Expect a mainly cytoplasmic pattern and evaluate whether positive cells fit the tissue context; the atlas reports High staining in testis preleptotene spermatocytes and no detection in adipocytes (HPA: tissue profile; HPA: supported cytosol). Strong signal confined to section edges, folds, or necrotic regions should trigger a processing review before biological interpretation (standard IHC practice). A diffuse nuclear-only pattern or sharp membrane rim also needs confirmation because the supported location is cytosolic and the protein has no transmembrane segment (HPA: supported cytosol; UniProt Q9NZI8 topology). Check no-primary and enzyme-block controls, then compare an adjacent section and a documented positive tissue before calling the unexpected region positive (standard IHC practice; HPA: testis profile).
Boster reagents
Best IGF2BP1 / Insulin-like growth factor 2 mRNA-binding protein 1 IHC Antibodies
Real IGF2BP1 IHC data show staining in human testis and lung adenocarcinoma paraffin sections (A02007-1 image captions). Mouse reactivity is listed, but no IF data are supplied (catalog).
Real IHC data
Anti-IGF2BP1 Antibody (C-term)
Cat # A02007-1
A02007-1 has IHC-P images from human testis and lung adenocarcinoma paraffin sections (A02007-1 image captions). It lists human and mouse reactivity and IHC-P as an application; no IF image is supplied (catalog).
Which to pick: Choose A02007-1 for tissue IHC-P: it is rabbit polyclonal, lists a 1:25 dilution, and its own captions describe paraffin sections with citrate retrieval at pH 6 (catalog; A02007-1 image captions). Its captions describe fixation as both paraformaldehyde and formaldehyde (A02007-1 image captions). No listed IF/ICC application or IF image supports an IF/ICC choice; for cross-species work, A02007-1 lists human and mouse reactivity, although its IHC images show human samples only (catalog; A02007-1 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NZI8 (IF2B1_HUMAN, Insulin-like growth factor 2 mRNA-binding protein 1).
- Human Protein Atlas. IGF2BP1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. IGF2BP1 subcellular location (ICC-IF): Mainly localized to the cytosol..
- Human Protein Atlas. IGF2BP1 antibody validation summary (2 antibodies).
- MicroRNAs as potential indicators of the development and progression of uterine leiomyoma. PloS one 2022 — PMC9154092.
- Differences in the expression profiles of m6A-related genes and the prognostic role in immunotherapy for lung adenocarcinoma patients. Frontiers in oncology 2026 — PMC12967950.
- IGF2BP1/HMOX1 Mediates Fate Determinations of Human Spermatogonial Stem Cells and Male Infertility via an m6A-Dependent Manner. Research (Washington, D.C.) 2025 — PMC12635413.
- Expression of microRNA-150 and its Target Gene IGF2BP1 in Human Osteosarcoma and their Clinical Implications. Pathology oncology research : POR 2019 — PMC6449306.
- PubMed PMID:16625196 — UniProt-cited evidence.
- PubMed PMID:9891060 — UniProt-cited evidence.
- PubMed PMID:16356927 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IGF2BP1 (Insulin-like growth factor 2 mRNA-binding protein 1). Expected localisation Cytoplasmic staining in testis and ovary (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A02007-1). Validated IHC antibody: A02007-1 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Preleptotene spermatocytes); negative Adipose tissue. IHC-P workflow on paraffin sections. Protocol steps: 1. Sample: Formaldehyde-fixed, paraffin-embedded human testis tissue (datasheet A02007-1); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A02007-1); 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 (datasheet A02007-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-IGF2BP1, 1:25 (datasheet A02007-1); 9. Primary incubation: 1 hours at 37°C (datasheet A02007-1). Watch for: Fixation: Use formaldehyde-fixed paraffin sections (selected-SKU IHC image A02007-1); keep fixation consistent (standard IHC practice; not target-specific); Caveat: Signal varies by tissue and cell type (HPA tissue IHC); Isoform / epitope: 2 isoforms ; verify C-terminal epitope coverage (UniProt; datasheet A02007-1); Regulation: Embryonic; expressed in some cancers (UniProt). Evidence: UniProt Q9NZI8, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol., Datasheet A02007-1, PMC9154092, PMC12967950. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IGF2BP1/immunohistochemistry-ihc-guide.