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- Table of Contents
HNRNPC / Heterogeneous nuclear ribonucleoproteins C1/C2 · Western blot design guide
Design a Western Blot for HNRNPC
Real validated HNRNPC Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HNRNPC 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
HNRNPC 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 | ~33.7 kDa | |
| Observed band | ~38 and 41 kDa | |
| Gel | 10% (catalog A02726-1) | |
| 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 Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated HNRNPC Western Blot Protocols
The A02726-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human HepG2, human K562 (catalog A02726-1) |
| Gel % | 10% (catalog A02726-1) |
| Load | 30 ug; reducing conditions (catalog A02726-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02726-1) |
| Membrane | nitrocellulose membrane (catalog A02726-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02726-1) |
| Primary antibody | A02726-1 · 1:1000 (catalog A02726-1) |
| Primary incubation | overnight at 4°C (catalog A02726-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:1000 (catalog A02726-1) |
| Secondary incubation | 1.5 hour at RT (catalog A02726-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02726-1) |
| Detection | ECL (catalog A02726-1) |
Section 2
What Is the Expected HNRNPC Western Blot Band Size?
HNRNPC is predicted at 33.7 kDa, while antibody QC reports ~38 and 41 kDa bands; the cause of the difference is not established.
| Bands at ~38 and 41 kDa | Empirical HNRNPC signals in whole-cell lysates; their assignment to specific isoforms is unconfirmed |
| A single band near 38 kDa | Matches one reported HNRNPC band; confirm its identity with an independent control |
| A single band near 41 kDa | Matches one reported HNRNPC band; confirm its identity with an independent control |
| A band near 33.7 kDa | Near the UniProt predicted mass; its identity requires confirmation |
| Additional discrete bands | HNRNPC has four named isoforms, but their individual migration positions are unknown |
💡Expected HNRNPC appearanceUniProt predicts 33.7 kDa, while antibody QC shows bands at ~38 and 41 kDa in whole-cell lysates; their difference from the prediction and isoform identities are unestablished, so confirm identity with independent controls.
| UniProt predicted mass | 33.7 kDa is the sequence-based reference, while observed bands are ~38 and 41 kDa |
| Isoform C1 | May migrate differently from other isoforms; its individual band position is unknown |
| Isoform C2 | May migrate differently from other isoforms; its individual band position is unknown |
| Isoforms 3 and 4 | May add band positions, but their relative sizes and migration are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear HNRNPC may be poorly recovered or diluted in whole-cell lysate | Check nuclear recovery and compare with a nuclear-enriched fraction |
| Band higher than expected | The reported ~38 and 41 kDa bands exceed the 33.7 kDa prediction for an undetermined reason | Compare with the reported band positions and confirm identity by knockdown or an independent antibody |
| Band lower than expected | A lower band could be a different isoform or a fragment; its identity is unestablished | Check sample integrity and test band identity with an independent antibody |
| Multiple bands | Bands at ~38 and 41 kDa are reported; four isoforms exist, but band assignments are unknown | Compare with the reported pattern and use knockdown or isoform-specific controls |
| Weak or no signal | Nuclear protein recovery or detection may be insufficient | Check nuclear recovery and include a positive lysate from a reported blot |
| Fragments below expected size | Sample degradation could produce fragments; no fragment size is established | Compare fresh, protease-inhibited lysate and confirm signal identity |
Experiment evidence
Western blot images showing HNRNPC band
A02726-1 · HNRNPC
M02726 · HNRNPC
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HNRNPC Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HNRNPC in Western blot, you can use adipose tissue lysate.
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.
⚠️Feasibility: No Not-detected tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for HNRNPC
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 |
|---|---|---|---|---|
| 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 → |
Section 3
Advanced HNRNPC Western Blot Tips
Deeper troubleshooting and optimisation questions for HNRNPC, answered from its protein features.
How should HNRNPC 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 can HNRNPC isoforms affect band interpretation?
Isoforms · Four isoforms are listed: C2, C1, 3 and 4. Relative to the supplied canonical sequence, C1 and 4 lack residues 108–120; isoform 4 also lacks 153–195; and isoform 3 replaces 39–119 with N. These sequence differences make isoforms relevant when assigning bands, but the supplied features do not assign identities to the 38 and 41 kDa bands.
No. In the supplied canonical coordinates, residues 108–120 are missing from C1 and 4, affecting listed sites 109, 113 and 115. Isoform 4 also lacks 153–195, affecting sites 162 and 166. Isoform 3 replaces 39–119 with N, affecting sites in that interval. Check the isoform sequence before interpreting a site-specific signal.
No. In the supplied canonical coordinates, residues 108–120 are missing from C1 and 4, affecting listed sites 109, 113 and 115. Isoform 4 also lacks 153–195, affecting sites 162 and 166. Isoform 3 replaces 39–119 with N, affecting sites in that interval. Check the isoform sequence before interpreting a site-specific signal.
Which HNRNPC phosphorylation sites are listed?
PTM · In the supplied UniProt coordinate convention, threonine 109 and serines 113, 115, 121, 162, 166, 233, 238, 239, 241, 253, 260, 299 and 306 are listed as phosphorylated. Match any antibody or paper numbering to this convention before comparing sites. Site presence does not establish a visible band shift.
The supplied UniProt features list N-acetylalanine at position 2 and alternate N6-acetyllysine at position 176, using canonical coordinates. Position 176 falls within the segment missing from isoform 4. These annotations do not demonstrate that acetylation causes either observed band.
The supplied UniProt features list N-acetylalanine at position 2 and alternate N6-acetyllysine at position 176, using canonical coordinates. Position 176 falls within the segment missing from isoform 4. These annotations do not demonstrate that acetylation causes either observed band.
Does this guide establish induction of HNRNPC?
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 HNRNPC?
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 A02726-1 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 the two observed HNRNPC bands be quantified?
Quantitation · Measure the approximately 38 and 41 kDa bands separately and state which signal is being compared. The four listed isoforms make band identity relevant, but the supplied features do not establish which isoform occupies either band. Combine band intensities only after establishing that both signals represent the intended HNRNPC measurement.
Why are HNRNPC bands observed near 38 and 41 kDa?
Interpretation · The supplied predicted mass is 33.7 kDa, while the observed bands are about 38 and 41 kDa. HNRNPC has four listed isoforms and multiple modifications, but these features alone do not explain the apparent masses or establish which band represents which isoform.
Compare them with the listed isoform sequence changes and the expected nuclear location. HNRNPC has no listed signal peptide, propeptide or glycosylation sites, so those features do not support assigning an extra band. Its phosphorylation and acetylation annotations also cannot, by themselves, identify or explain an unexpected band.
Compare them with the listed isoform sequence changes and the expected nuclear location. HNRNPC has no listed signal peptide, propeptide or glycosylation sites, so those features do not support assigning an extra band. Its phosphorylation and acetylation annotations also cannot, by themselves, identify or explain an unexpected band.
Boster reagents
HNRNPC 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-HNRNPC Antibody
Cat # A02726-1
Real WB data
Anti-hnRNP C1/C2 Rabbit Monoclonal Antibody
Cat # M02726
A02726-1 is a rabbit polyclonal antibody; M02726 is a rabbit monoclonal antibody. Both have WB images using human, rat, and mouse lysates. Reported bands are 38–41 kDa and approximately 40 kDa, respectively, above their stated expected sizes. No publication evidence is supplied.
Which to pick: Both list human, mouse, and rat reactivity and have WB images. Choose by the closest tested specimen: A02726-1 includes MCF-7, C6, Neuro-2a, and HEPA1-6 cells; M02726 includes Jurkat and NIH/3T3 cells plus rat and mouse brain tissue. Both used a 1:1000 primary dilution.
Source: BosterBio HNRNPC gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P07910.
- Human Protein Atlas. HNRNPC tissue expression.
- PMC13477384 — target-verified WB comparison
- PMC7296632 — target-verified WB comparison
- PMC13122172 — target-verified WB comparison