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- Table of Contents
HOXC8 / Homeobox protein Hox-C8 · Western blot design guide
Design a Western Blot for HOXC8
Source-linked HOXC8 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HOXC8 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
HOXC8 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 | ~27.8 kDa | |
| Observed band | ~35 kDa | |
| Gel | 5–20% (catalog A06655-1) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Observed size differs | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked HOXC8 Western Blot Protocol Options
The A06655-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human COLO-320, human U20S, human A549, rat brain, mouse brain, mouse ovary (catalog A06655-1) |
| Gel % | 5–20% (catalog A06655-1) |
| Load | 50ug; reducing conditions (catalog A06655-1) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A06655-1) |
| Membrane | Nitrocellulose membrane (catalog A06655-1) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A06655-1) |
| Primary antibody | A06655-1 · 0.5 μg/mL (catalog A06655-1) |
| Primary incubation | overnight at 4°C (catalog A06655-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06655-1) |
| Secondary incubation | 1.5 hour at RT (catalog A06655-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06655-1) |
| Detection | ECL (catalog A06655-1) |
Section 2
What Is the Expected HOXC8 Western Blot Band Size?
HOXC8 is predicted at 27.8 kDa, while antibody QC reports approximately 35 kDa; the cause of the difference is not established.
| Band near 35 kDa | Matches the reported HOXC8 band; confirm identity with an antibody control. |
| Band near 27.8 kDa | Near the predicted mass, but identity requires confirmation. |
| Faint band in whole-cell lysate | Nuclear HOXC8 may be diluted among whole-cell proteins. |
| Stronger band in a nuclear fraction | Consistent with HOXC8 nuclear localization. |
💡Expected HOXC8 appearanceHOXC8 has a predicted mass of 27.8 kDa, while antibody QC reports a band near 35 kDa; the cause of the difference is unknown, so confirm band identity with appropriate controls.
| UniProt predicted mass | The sequence predicts 27.8 kDa, below the reported band near 35 kDa. |
| Sequence-derived molecular weight | The calculated mass is 27,755 Da, equivalent to approximately 27.8 kDa. |
| Full-length sequence prediction | The 242-residue sequence underlies the 27.8 kDa prediction. |
| Predicted mass versus apparent migration | The 27.8 kDa prediction alone does not explain migration near 35 kDa. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear HOXC8 may be dilute in whole-cell lysate. | Test a nuclear-enriched fraction alongside the lysate. |
| Band higher than expected | The reported band is near 35 kDa, above the 27.8 kDa prediction; the cause is unknown. | Compare with the reported blot and confirm identity using antibody controls. |
| Band lower than expected | No listed HOXC8 feature explains a lower band. | Check sample integrity and confirm identity with an independent antibody. |
| Multiple bands | The supplied features do not establish multiple HOXC8 bands. | Check antibody specificity with a HOXC8 depletion control. |
| Weak or no signal | The nuclear target may be underrepresented in the sample. | Check loading and test a nuclear-enriched fraction. |
| Fragments below expected size | Sample degradation is possible; no cleavage feature is listed. | Prepare fresh lysate with protease inhibitors and compare bands. |
Experiment evidence
Western blot images showing HOXC8 band
A06655-1 · HOXC8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HOXC8 Western blot
🧪For positive controls for HOXC8 in Western blot, you can use no HPA-IHC candidate positive sample because no positive tissue or cell is supplied.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: With no HPA expression data, a positive sample cannot be selected and a tissue negative cannot be validated.
HPA tissue expression evidence for HOXC8
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced HOXC8 Western Blot Tips
Deeper troubleshooting and optimisation questions for HOXC8, answered from its protein features.
How should HOXC8 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 annotated HOXC8 isoforms explain multiple bands?
Isoforms · Only one HOXC8 isoform is listed, with no alternative sequence. Do not assign additional bands to annotated isoforms on this evidence alone; verify their identity independently.
Can annotated modifications explain HOXC8's apparent mass?
PTM · The supplied UniProt features list no modified residues or glycosylation sites. They therefore provide no specific modification to test as the cause of the approximately 35 kDa band. The absence of an annotation does not establish why the band migrates there.
Does this guide establish induction of HOXC8?
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 HOXC8 Western blot?
Transfer · HOXC8 is predicted to be 27.8 kDa and is observed near 35 kDa. Use transfer conditions suited to that size range, then check the membrane and gel for transfer efficiency. The supplied features do not specify a required transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06655-1 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 HOXC8 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.
Why might HOXC8 appear near 35 kDa instead of 27.8 kDa?
Interpretation · HOXC8 has a predicted mass of 27.8 kDa, while the supplied Western blot observation is approximately 35 kDa. These data do not establish the cause of the difference. Compare the band with a molecular weight ladder and confirm its identity before assigning it to HOXC8.
HOXC8 is annotated as nuclear. Include a nuclear fraction when assessing its abundance, and keep the fractionation method consistent across samples used for comparison.
Confirm an unexpected band before identifying it as HOXC8. The supplied features list one isoform and no annotated modified residues or glycosylation sites. HOXC8 interacts with HOMEZ and forms a DNA-binding heterodimer with PBX1, but those interactions alone do not establish the identity of another Western blot band.
HOXC8 is annotated as nuclear. Include a nuclear fraction when assessing its abundance, and keep the fractionation method consistent across samples used for comparison.
Confirm an unexpected band before identifying it as HOXC8. The supplied features list one isoform and no annotated modified residues or glycosylation sites. HOXC8 interacts with HOMEZ and forms a DNA-binding heterodimer with PBX1, but those interactions alone do not establish the identity of another Western blot band.
Boster reagents
HOXC8 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-HOXC8 Antibody Picoband®
Cat # A06655-1
The catalog reports one anti-HOXC8 antibody, A06655-1, with reported human, mouse, and rat reactivity. Its Western blot image shows an approximately 35 kDa band in the listed cell and tissue lysates. Evidence is limited to the supplied product blot; no publication evidence is provided.
Which to pick: A06655-1 is the only listed option and has a Western blot image using human cell, rat brain, mouse brain, and mouse ovary lysates. The reported conditions used 50 µg of lysate per lane and 0.5 µg/mL primary antibody.
Source: BosterBio HOXC8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P31273.
- Human Protein Atlas. HOXC8 tissue expression.
- PMC9076220 — target-verified WB comparison
- PMC10598528 — target-verified WB comparison
- PMC5833702 — target-verified WB comparison