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- Table of Contents
HNRNPU / Heterogeneous nuclear ribonucleoprotein U · Western blot design guide
Design a Western Blot for HNRNPU
Real validated HNRNPU Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HNRNPU 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
HNRNPU 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 | ~90.6 kDa | |
| Gel | 8–10% (standard starting point) | |
| 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 | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated HNRNPU Western Blot Protocols
The M03691 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | K562 cell lysate (catalog M03691) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M03691; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| 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 HNRNPU Western Blot Band Size?
HNRNPU is predicted at 90.6 kDa; isoforms and modifications are documented, but their effects on blot migration are unproven.
| Band near 90.6 kDa | consistent with the predicted HNRNPU mass; confirm identity with controls |
| Two bands at different positions | isoforms 1 and 2 are possible, but distinct migration is unverified |
| Weak band in a cytoplasmic fraction | consistent with HNRNPU's nuclear localization |
| Band away from 90.6 kDa | its identity and cause of altered migration are unestablished |
💡Expected HNRNPU appearanceHNRNPU has a predicted mass of 90.6 kDa, but no empirical band size is supplied; confirm a candidate band with appropriate identity controls.
| Predicted molecular mass | places the unmodified sequence at 90.6 kDa |
| Isoform 1 | has no supplied isoform-specific mass or migration |
| Isoform 2 | has no supplied isoform-specific mass or migration |
| Isoforms 1 and 2 | may differ in size, but their relative migration is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | insufficient recovery of nuclear HNRNPU is possible | check nuclear enrichment and a positive-control lysate |
| Band higher than expected | the cause of altered migration is unestablished | compare with a molecular-weight marker and verify identity by HNRNPU depletion |
| Band lower than expected | the band could be an unverified isoform or fragment | verify identity by HNRNPU depletion and compare antibody epitopes |
| Multiple bands | isoforms 1 and 2 are possible, but separate bands are unverified | test which bands respond to HNRNPU depletion |
| Weak or no signal | nuclear HNRNPU may be poorly recovered | check nuclear protein recovery and use a positive-control lysate |
Experiment evidence
Western blot images showing HNRNPU band
M03691 · HNRNPU
A03691-3 · HNRNPU
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HNRNPU Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HNRNPU in Western blot, you can use adrenal gland tissue.
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 alongside the samples.
⚠️Feasibility: No tissue is reported as undetected, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for HNRNPU
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 → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Medium | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Medium | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced HNRNPU Western Blot Tips
Deeper troubleshooting and optimisation questions for HNRNPU, answered from its protein features.
How should HNRNPU 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.
Could HNRNPU isoforms produce separate bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 213–231 of the canonical sequence, so it may migrate differently. Check whether the antibody recognizes a region retained in both isoforms before assigning bands.
Which phosphorylation site is linked to PLK1?
PTM · UniProt lists phosphoserine at position 59 by PLK1. Use UniProt sequence numbering when comparing this site with antibody or paper coordinates. This annotation alone does not predict a detectable band shift.
UniProt lists acetylation, ADP-ribosylation, citrullination, phosphorylation and arginine methylation. For example, serine 187 is ADP-ribosylated, and arginine 739 has alternate methylation annotations, including dimethylation in A2780 ovarian carcinoma cells. These features identify possible molecular differences, not confirmed causes of extra bands.
UniProt lists acetylation, ADP-ribosylation, citrullination, phosphorylation and arginine methylation. For example, serine 187 is ADP-ribosylated, and arginine 739 has alternate methylation annotations, including dimethylation in A2780 ovarian carcinoma cells. These features identify possible molecular differences, not confirmed causes of extra bands.
Does this guide establish induction of HNRNPU?
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.
What transfer method to use for HNRNPU Western blot?
Transfer · Choose a transfer setup suitable for a protein near its predicted 90.6 kDa mass, and verify that protein in this size range transfers. The supplied features do not specify a membrane or transfer conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03691 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 HNRNPU 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.
Should HNRNPU appear exactly at its predicted 90.6 kDa?
Interpretation · Use 90.6 kDa as the calculated reference. The listed modifications and isoforms do not establish an apparent band position or prove a visible shift; no observed band is supplied.
HNRNPU is annotated as nuclear and can oligomerize through its ATPase domain and RNA-binding RGG-box region in an ATP-, chromatin-associated RNA- and transcription-dependent manner. Keep sample preparation and the band selected for quantitation consistent across samples.
Compare bands with the 90.6 kDa calculated reference and consider isoform 2, which lacks residues 213–231. HNRNPU also has numerous annotated modifications and conditional oligomerization. None of these features alone identifies an unexpected band; check antibody recognition and sample preparation before assigning it.
HNRNPU is annotated as nuclear and can oligomerize through its ATPase domain and RNA-binding RGG-box region in an ATP-, chromatin-associated RNA- and transcription-dependent manner. Keep sample preparation and the band selected for quantitation consistent across samples.
Compare bands with the 90.6 kDa calculated reference and consider isoform 2, which lacks residues 213–231. HNRNPU also has numerous annotated modifications and conditional oligomerization. None of these features alone identifies an unexpected band; check antibody recognition and sample preparation before assigning it.
Boster reagents
HNRNPU 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-hnRNP U Monoclonal Antibody
Cat # M03691
Real WB data
Anti-hnRNP U/p120/HNRNPU Antibody Picoband®
Cat # A03691-3
Two the supplier anti-HNRNPU antibodies list human, mouse, and rat reactivity and have Western blot images. M03691 shows K562 lysate; A03691-3 shows human cell, rat cell and tissue, and mouse cell and tissue lysates. A03691-3 reports a 120 kDa band versus an expected 90 kDa.
Which to pick: Choose M03691 for a K562 example. Choose A03691-3 for documented WB examples across human, rat, and mouse samples, while accounting for its reported 120 kDa band. The images document those specific samples, not every sample from each listed species.
Source: BosterBio HNRNPU gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q00839.
- Human Protein Atlas. HNRNPU tissue expression.
- PMC12686487 — target-verified WB comparison
- PMC11705778 — target-verified WB comparison
- PMC8819945 — target-verified WB comparison