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IGF2BP2 / Insulin-like growth factor 2 mRNA-binding protein 2 · Western blot design guide
Design a Western Blot for IGF2BP2
Real validated IGF2BP2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-IGF2BP2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
IGF2BP2 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~66.1 kDa | |
| Observed band | ~66 kDa | |
| Gel | 10% (catalog A02010-4) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 6 isoform(s) |
Section 1
Real Curated IGF2BP2 Western Blot Protocols
The A02010-4 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human CACO-2, human HepG2, human K562 (catalog A02010-4) |
| Gel % | 10% (catalog A02010-4) |
| Load | 30 ug; reducing conditions (catalog A02010-4) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02010-4) |
| Membrane | nitrocellulose membrane (catalog A02010-4) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02010-4) |
| Primary antibody | A02010-4 · 0.5 μg/mL (catalog A02010-4) |
| Primary incubation | overnight at 4°C (catalog A02010-4) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02010-4) |
| Secondary incubation | 1.5 hour at RT (catalog A02010-4) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02010-4) |
| Detection | ECL (catalog A02010-4) |
Section 2
What Is the Expected IGF2BP2 Western Blot Band Size?
IGF2BP2 is predicted at 66.1 kDa and observed near 66 kDa; the supplied evidence does not establish a cause for the small difference.
| Band near 66 kDa | Consistent with the observed IGF2BP2 band; confirm identity with antibody controls |
| Additional lower band | A shorter isoform or alternative initiation is possible; identity needs confirmation |
| Several distinct bands | Isoforms 1–6 are annotated, but their migration differences are unknown |
| Weak band in soluble lysate | Recovery may vary because IGF2BP2 localizes to cytoplasmic mRNP granules |
💡Expected IGF2BP2 appearanceIGF2BP2 has a predicted mass of 66.1 kDa and an observed band near 66 kDa; confirm band identity with antibody specificity controls.
| UniProt predicted mass | Places the predicted protein near 66.1 kDa, consistent with the observed ~66 kDa band |
| Isoforms 1–6 | Could differ in size, but their masses and migration differences are not supplied |
| Phosphoserine at residue 11 | May affect mobility, but no visible shift is established |
| Phosphoserines at residues 162 and 164 | May affect mobility, but no visible shift is established |
| Phosphothreonine annotation at residue 550 | May affect mobility, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | IGF2BP2 recovery may vary with its nuclear and cytoplasmic granule localization | Check fraction recovery and a positive lysate |
| Band higher than expected | RNA-dependent oligomerization is possible, but persistence during SDS-PAGE is unproven | Check denaturation and confirm band identity |
| Band lower than expected | A shorter isoform or alternative initiation is possible | Confirm identity with an independent antibody or knockdown |
| Multiple bands | Isoforms 1–6 are annotated, but distinct bands are not established | Compare with knockdown and an independent antibody |
| Weak or no signal | Expression or extraction from nuclear and cytoplasmic granules may vary | Check loading, extraction, and a positive lysate |
| Fragments below expected size | Fragment identity is not established by the supplied features | Check sample integrity and test with an independent antibody |
Experiment evidence
Western blot images showing IGF2BP2 band
A02010-4 · IGF2BP2
M02010-1 · IGF2BP2
M02010-2 · IGF2BP2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for IGF2BP2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for IGF2BP2 in Western blot, you can use adrenal gland lysate, supported by high HPA expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside samples.
⚠️Feasibility: No HPA not-detected tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for IGF2BP2
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Breast | glandular cells | Low | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Low | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced IGF2BP2 Western Blot Tips
Deeper troubleshooting and optimisation questions for IGF2BP2, answered from its protein features.
How should IGF2BP2 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Which IGF2BP2 isoforms could produce bands of different sizes?
Isoforms · Six isoforms are listed. Isoforms 2 and 5 lack residues 358–400; isoform 6 lacks residues 1–68. Isoforms 3, 4 and 5 replace residues 1–80 with a shorter sequence, and isoform 4 also changes residue 113. These sequence differences could affect migration, but band positions are not supplied.
Check the antibody epitope against the isoform sequences. Residues 1–80 are replaced in isoforms 3, 4 and 5; residues 1–68 are absent in isoform 6; and residues 358–400 are absent in isoforms 2 and 5. An epitope within an altered region may not detect every isoform.
Decide whether to quantify one band or all detected IGF2BP2 bands, then apply that rule consistently. Six isoforms are listed, with several sequence changes that may affect migration or antibody recognition. The supplied features do not identify which isoform produces a given band.
Check the antibody epitope against the isoform sequences. Residues 1–80 are replaced in isoforms 3, 4 and 5; residues 1–68 are absent in isoform 6; and residues 358–400 are absent in isoforms 2 and 5. An epitope within an altered region may not detect every isoform.
Decide whether to quantify one band or all detected IGF2BP2 bands, then apply that rule consistently. Six isoforms are listed, with several sequence changes that may affect migration or antibody recognition. The supplied features do not identify which isoform produces a given band.
Which annotated modifications matter when interpreting IGF2BP2 bands?
PTM · The supplied UniProt coordinates list phosphoserine at 11, 162 and 164, plus an annotation labeled ‘Phosphothreonine N-acetylmethionine’ at 550. Keep that combined label as supplied; it does not establish a separate acetylation coordinate. These annotations alone do not demonstrate a visible band shift.
Does this guide establish induction of IGF2BP2?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for IGF2BP2?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02010-4 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should IGF2BP2 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Does the observed 66 kDa band match predicted IGF2BP2 mass?
Interpretation · Yes. The observed band is approximately 66 kDa, close to the predicted 66.1 kDa. The supplied features do not establish a cause for any small difference in apparent mass.
IGF2BP2 can form RNA-dependent homooligomers and heterooligomers with IGF2BP1 and IGF2BP3. This makes complexes a possibility to investigate, but the supplied features do not establish that a higher Western-blot band is an oligomer.
Yes. IGF2BP2 is listed in the nucleus and cytoplasm, including P-bodies, stress granules and cytoplasmic mRNP granules. Compare like fractions when quantifying samples; a change in a fraction's signal alone does not establish a change in total IGF2BP2.
IGF2BP2 can form RNA-dependent homooligomers and heterooligomers with IGF2BP1 and IGF2BP3. This makes complexes a possibility to investigate, but the supplied features do not establish that a higher Western-blot band is an oligomer.
Yes. IGF2BP2 is listed in the nucleus and cytoplasm, including P-bodies, stress granules and cytoplasmic mRNP granules. Compare like fractions when quantifying samples; a change in a fraction's signal alone does not establish a change in total IGF2BP2.
Boster reagents
IGF2BP2 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-IGF2BP2 Antibody Picoband®
Cat # A02010-4
Real WB data
Anti-IMP2 Rabbit Monoclonal Antibody
Cat # M02010-1
Real WB data
Anti-IGF2BP2 Rabbit Monoclonal Antibody
Cat # M02010-2
Three the supplier IGF2BP2 antibodies list human, mouse, and rat reactivity and have WB images. A02010-4 and M02010-1 report approximately 66 kDa bands in specified lysates; M02010-2 has only a brief 293T lysate caption. These images do not establish performance in every sample.
Which to pick: Choose by the closest reported sample: A02010-4 includes CACO-2, C6, and mouse testis; M02010-1 includes SH-SY5Y and rat and mouse heart; M02010-2 shows 293T lysate, with fewer reported WB details. All three list human, mouse, and rat reactivity.
Source: BosterBio IGF2BP2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9Y6M1.
- Human Protein Atlas. IGF2BP2 tissue expression.
- PMC12721180 — target-verified WB comparison
- PMC12500958 — target-verified WB comparison
- PMC6921559 — target-verified WB comparison