This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ING4 / Inhibitor of growth protein 4 · Western blot design guide
Design a Western Blot for ING4
Source-linked ING4 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ING4 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
ING4 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~28.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 8 isoform(s) |
Section 1
Source-Linked ING4 Western Blot Protocol Options
The A02664-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts from Jurkat, K562 cells, (catalog A02664-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A02664-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:20000 (catalog A02664-1) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected ING4 Western Blot Band Size?
ING4 has a predicted monomer mass of 28.5 kDa; isoforms and homodimerization could affect band patterns, but no empirical migration is supplied.
| Band near 28.5 kDa | consistent with the predicted ING4 monomer; confirm its identity |
| Band near 57 kDa | could reflect a homodimer that persists during electrophoresis |
| Several bands near the monomer region | could reflect ING4 splice isoforms; their migration is not established |
| Weak whole-cell band with stronger nuclear-fraction signal | consistent with ING4 nuclear localization |
💡Expected ING4 appearanceING4 has a predicted monomer mass of 28.5 kDa, but no empirical band size is supplied; confirm any candidate band with nuclear enrichment and an ING4-specific identity control.
| Predicted ING4 monomer mass | provides a 28.5 kDa reference, not a measured migration |
| Homodimer formation | could yield a band near twice the monomer size if the dimer persists during electrophoresis |
| Splice isoforms 1, 2, 3, and 4 | may differ in apparent size; their individual masses and migration are not supplied |
| Splice isoforms 5, 6, 7, and 8 | may differ in apparent size; distinct resolved bands are not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear ING4 may be diluted in whole-cell lysate | check a nuclear fraction and validate the candidate band with ING4 knockdown |
| Band higher than expected | an ING4 homodimer may persist during electrophoresis | compare denaturing conditions and test band identity with ING4 knockdown |
| Band lower than expected | a splice isoform is possible, but its mass is unknown | compare isoform-specific expression and confirm the band with ING4 knockdown |
| Multiple bands | ING4 has eight annotated splice isoforms, though distinct migration is unproven | use isoform-specific controls and ING4 knockdown to identify the bands |
| Weak or no signal | nuclear ING4 may be poorly represented in the sample | enrich the nuclear fraction and verify fraction quality |
Experiment evidence
Western blot images showing ING4 band
A02664-1 · ING4
A02664-1 · ING4
A02664-2 · ING4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ING4 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ING4 in Western blot, you can use adrenal gland tissue, which HPA scores as medium expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA scores the positive tissue as medium expression, so the ING4 band may be modest.
HPA tissue expression evidence for ING4
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 | Medium | Protein (IHC) | HPA → |
| Cervix | glandular cells | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Medium | Protein (IHC) | HPA → |
| Kidney | cells in tubules | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Colon | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ING4 Western Blot Tips
Deeper troubleshooting and optimisation questions for ING4, answered from its protein features.
What band size should I expect for ING4?
Band shift · The supplied predicted mass is 28.5 kDa. No observed band size is available. Use 28.5 kDa as a reference, but do not assign a band by size alone: ING4 has eight annotated isoforms, and its modification features do not establish a visible shift.
Which ING4 isoforms could produce smaller bands?
Isoforms · Isoform 3 replaces canonical residues 13–37 with one residue; isoform 8 replaces residues 168–249 with a shorter sequence. Both could differ substantially in size. Other isoforms have shorter changes around residues 128–132. These are UniProt canonical coordinates, and the supplied features do not establish their observed band positions.
Check the antibody epitope against the eight annotated isoforms. Isoform 3 changes residues 13–37, isoforms 2, 4, 5, 6, and 7 change the region around 128–132, and isoform 8 changes residues 168–249. An antibody targeting an altered region may detect isoforms differently.
Check the antibody epitope against the eight annotated isoforms. Isoform 3 changes residues 13–37, isoforms 2, 4, 5, 6, and 7 change the region around 128–132, and isoform 8 changes residues 168–249. An antibody targeting an altered region may detect isoforms differently.
Could acetylation explain an ING4 band shift?
PTM · UniProt lists N6-acetyllysine at canonical positions 112, 127, 129, 146, 148, and 156; position 166 also includes an N6-acetyllysine annotation. These features identify candidate modified residues but do not demonstrate a visible shift or explain a difference from the predicted 28.5 kDa mass.
Does this guide establish induction of ING4?
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 ING4?
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 A02664-1 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 I quantify ING4 across samples?
Quantitation · ING4 is annotated as nuclear and has eight isoforms. Compare the same sample fraction across lanes and decide whether to quantify one band or all detected isoform bands. Use an antibody epitope shared by the isoforms you intend to measure, since several isoforms alter the N terminus, residues 128–132, or the C terminus.
Where is ING4 citrullination annotated?
Interpretation · The supplied UniProt features list citrulline at canonical positions 133 and 166. Position 166 also carries N6-acetyllysine annotations in the supplied record. Keep those annotations and coordinates explicit when comparing results; the features alone do not establish a separate band.
Compare each band with the 28.5 kDa prediction and the annotated isoform changes, particularly those in isoforms 3 and 8. ING4 is also annotated as a homodimer and a component of the HBO1 complex, but those features do not establish that a higher band represents an intact complex on a Western blot. Confirm band identity before assigning it to an isoform or modification.
Compare each band with the 28.5 kDa prediction and the annotated isoform changes, particularly those in isoforms 3 and 8. ING4 is also annotated as a homodimer and a component of the HBO1 complex, but those features do not establish that a higher band represents an intact complex on a Western blot. Confirm band identity before assigning it to an isoform or modification.
Boster reagents
ING4 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-Inhibitor of growth protein 4 ING4 Antibody
Cat # A02664-1
Real WB data
Anti-Inhibitor of growth protein 4 ING4 Antibody
Cat # A02664-2
Two anti-ING4 antibodies have WB images. A02664-1 shows Jurkat and K562 nuclear extracts; A02664-2 shows extracts from unspecified cell lines with reported blot conditions. No publication evidence is supplied.
Which to pick: Choose A02664-1 if you need stated rat reactivity or a WB example using Jurkat or K562 nuclear extracts. A02664-2 lists human and mouse reactivity and reports a 1:1000 primary dilution. Both have WB images; the examples do not establish validation across all listed species.
Source: BosterBio ING4 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UNL4.
- Human Protein Atlas. ING4 tissue expression.
- PMC6895402 — target-verified WB comparison
- PMC6367549 — target-verified WB comparison
- PMC11226078 — target-verified WB comparison