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- Table of Contents
HNRNPK / Heterogeneous nuclear ribonucleoprotein K · Western blot design guide
Design a Western Blot for HNRNPK
Real validated HNRNPK Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HNRNPK 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
HNRNPK 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 | ~51 kDa | |
| Observed band | ~60 kDa | |
| Gel | 5–20% (catalog A01793-2) | |
| Positive control | Adipose tissue (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 C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated HNRNPK Western Blot Protocols
The A01793-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human MCF-7, human MOLT-4 (catalog A01793-2) |
| Gel % | 5–20% (catalog A01793-2) |
| Load | 30 ug; reducing conditions (catalog A01793-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01793-2) |
| Membrane | nitrocellulose membrane (catalog A01793-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01793-2) |
| Primary antibody | A01793-2 · 0.5 μg/mL (catalog A01793-2) |
| Primary incubation | overnight at 4°C (catalog A01793-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01793-2) |
| Secondary incubation | 1.5 hour at RT (catalog A01793-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01793-2) |
| Detection | ECL (catalog A01793-2) |
Section 2
What Is the Expected HNRNPK Western Blot Band Size?
HNRNPK is predicted at 51 kDa and observed near 60 kDa; the cause of the difference is not established.
| Band near 60 kDa | Matches the empirical HNRNPK band; confirm identity with antibody controls. |
| Band near 51 kDa | Near the predicted sequence mass; identity requires confirmation. |
| Several bands at different positions | Isoforms 1, 2 and 3 are reported, but distinct migration has not been established. |
| Bands in nuclear and cytoplasmic fractions | Consistent with HNRNPK localization in both compartments. |
💡Expected HNRNPK appearanceUniProt predicts 51 kDa, while antibody QC shows a band near 60 kDa; the cause of the difference is unestablished, so confirm band identity with ordinary antibody controls.
| UniProt predicted mass | The sequence predicts 51 kDa, while the empirical band is near 60 kDa; the cause of the difference is unestablished. |
| Isoforms 1, 2 and 3 | Alternative sequences could differ in mass, but their individual masses and migration are not supplied. |
| Phosphorylation at Ser36, Thr39, Ser75, Ser116 and Ser214 | May affect migration; no visible shift is established. |
| Acetylation at Met1, Lys34 and Lys198 | May affect migration; no visible shift is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | HNRNPK occurs in the nucleoplasm, and its recovery from the lysate is uncertain. | Check a positive-control lysate and nuclear protein recovery. |
| Band higher than expected | The empirical band is near 60 kDa versus a predicted 51 kDa; its migration difference is unexplained. | Compare with the reported 60 kDa band and confirm identity by HNRNPK knockdown. |
| Band lower than expected | An alternative isoform is possible, but its migration is unreported. | Check antibody specificity and confirm the band by HNRNPK knockdown. |
| Multiple bands | Isoforms 1, 2 and 3 or modified forms are possible, but distinct bands are unestablished. | Use HNRNPK knockdown to identify specific bands. |
| Weak or no signal | Poor recovery of nucleoplasmic HNRNPK is possible. | Check nuclear protein recovery and run a positive-control lysate. |
Experiment evidence
Western blot images showing HNRNPK band
A01793-2 · HNRNPK
M01793 · HNRNPK
M01793 · HNRNPK
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HNRNPK Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HNRNPK in Western blot, you can use adipose tissue lysate, which shows high expression in HPA.
Positive control: Adipose tissue (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: HPA lists no tissue with undetected expression, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for HNRNPK
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | Medium | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Medium | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced HNRNPK Western Blot Tips
Deeper troubleshooting and optimisation questions for HNRNPK, answered from its protein features.
How should HNRNPK 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.
How could HNRNPK isoforms affect band interpretation?
Isoforms · UniProt lists three isoforms. Isoform 3 lacks canonical residues 111–134; isoforms 2 and 3 replace canonical SGKFF at 459–463 with ADVEGF. The deletion could change mass, but distinct bands are not guaranteed. Compare samples using an antibody whose epitope is known.
Which phosphorylation sites could inform follow-up on HNRNPK bands?
PTM · UniProt lists S36, T39, S75, S116, S214, S216, S284, S379, Y380, and S420 in canonical coordinates. Compare matched samples with and without phosphatase treatment if testing a phosphorylation-related band pattern. S116 lies within the segment missing from isoform 3. A listed site does not by itself establish a visible shift.
In canonical UniProt coordinates, HNRNPK has N-acetylmethionine at 1; acetyllysine at 34, 198, and 405; succinyllysine at 219; and methylarginine at 316 and 377. UniProt marks modifications at 34, 219, and 405 as alternate. These annotations identify possible states, but do not establish which state produced a band.
In canonical UniProt coordinates, HNRNPK has N-acetylmethionine at 1; acetyllysine at 34, 198, and 405; succinyllysine at 219; and methylarginine at 316 and 377. UniProt marks modifications at 34, 219, and 405 as alternate. These annotations identify possible states, but do not establish which state produced a band.
Does this guide establish induction of HNRNPK?
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 HNRNPK?
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 A01793-2 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 HNRNPK be quantified across cellular fractions?
Quantitation · UniProt places HNRNPK in the nucleoplasm and cytoplasm, and also lists it at podosomes. Quantify comparable fractions across samples, with a suitable loading reference for each fraction. A change in one fraction need not represent a change in total HNRNPK.
Why might HNRNPK appear near 60 kDa rather than its predicted 51 kDa?
Interpretation · The supplied apparent band is ~60 kDa, while the predicted mass is 51 kDa. UniProt lists alternative isoforms and modifications, but their presence alone does not explain the difference or establish a visible shift. Verify the band with an HNRNPK-specific control before assigning it.
Check whether the epitope overlaps canonical residues 111–134, which are absent from isoform 3, or 459–463, which differ in isoforms 2 and 3. An antibody to either region may detect the isoforms differently. These coordinates use the supplied canonical UniProt sequence.
Compare them with the ~60 kDa observed band and the 51 kDa predicted mass. Check antibody epitope coverage against the three isoforms, particularly the isoform 3 deletion at canonical 111–134. The listed modifications offer follow-up targets, but neither isoform annotations nor modification sites alone identify an unexpected band.
Check whether the epitope overlaps canonical residues 111–134, which are absent from isoform 3, or 459–463, which differ in isoforms 2 and 3. An antibody to either region may detect the isoforms differently. These coordinates use the supplied canonical UniProt sequence.
Compare them with the ~60 kDa observed band and the 51 kDa predicted mass. Check antibody epitope coverage against the three isoforms, particularly the isoform 3 deletion at canonical 111–134. The listed modifications offer follow-up targets, but neither isoform annotations nor modification sites alone identify an unexpected band.
Boster reagents
HNRNPK 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 K/HNRNPK Antibody Picoband®
Cat # A01793-2
Real WB data
Anti-hnRNP K Rabbit Monoclonal Antibody
Cat # M01793
The catalog reports two anti-HNRNPK antibodies with WB images and reported human, mouse, and rat reactivity. Their captions show bands near 60 kDa in the specified cell and tissue lysates; these examples do not establish performance in every sample.
Which to pick: Choose A01793-2 for the polyclonal antibody with illustrated rat PC-12 and mouse NIH/3T3 lysates. Choose monoclonal M01793 if its illustrated human HepG2 or mouse thymus samples better match your experiment. Both have WB images spanning human, rat, and mouse samples.
Source: BosterBio HNRNPK gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P61978.
- Human Protein Atlas. HNRNPK tissue expression.
- PMC4150800 — target-verified WB comparison
- PMC12165811 — target-verified WB comparison
- PMC7969774 — target-verified WB comparison