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- Table of Contents
XRCC3 / DNA repair protein XRCC3 · Western blot design guide
Design a Western Blot for XRCC3
Source-linked XRCC3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-XRCC3 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
XRCC3 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 | ~37.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked XRCC3 Western Blot Protocol Options
The M01068-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog M01068-1) |
| Gel % | 12–15% (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 | M01068-1; 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 XRCC3 Western Blot Band Size?
XRCC3 is predicted at 37.9 kDa; its listed features do not demonstrate a different migration, and no empirical band size is supplied.
| Single band near 37.9 kDa in whole-cell lysate | consistent with predicted XRCC3 size if band identity is confirmed |
| Band near 37.9 kDa in a nuclear fraction | consistent with XRCC3 nuclear localization |
| Band near 37.9 kDa in a cytoplasmic fraction | consistent with XRCC3 cytoplasmic localization |
| Band near 37.9 kDa in a mitochondrial fraction | consistent with XRCC3 mitochondrial localization |
💡Expected XRCC3 appearanceUniProt predicts a 37.9 kDa XRCC3 band; no empirical band size is supplied, so confirm any candidate band with an XRCC3 depletion or knockout control.
| UniProt predicted molecular weight | places the expected full-length band near 37.9 kDa |
| UniProt mass in daltons | gives the same predicted mass as 37,850 Da |
| 346-residue XRCC3 sequence | defines the full-length protein used for the mass prediction |
| Predicted full-length XRCC3 reference | provides a size reference but does not establish actual gel migration |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | XRCC3 may be below detection or lost during extraction | check loading and assay a nuclear-enriched sample |
| Band higher than expected | band identity or apparent migration is unverified | compare with an XRCC3 depletion or knockout control |
| Band lower than expected | the band may be a fragment or unrelated signal | repeat with protease inhibitors and verify band identity |
| Multiple bands | additional bands are not explained by listed XRCC3 isoforms | identify which bands decrease after XRCC3 depletion |
| Weak or no signal | XRCC3 may be dilute across cellular compartments | check loading and test nuclear enrichment |
| Fragments below expected size | XRCC3 may have degraded during sample preparation | prepare fresh lysate with protease inhibitors |
Experiment evidence
Western blot images showing XRCC3 band
M01068-1 · XRCC3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for XRCC3 Western blot
🧪For positive controls for XRCC3 in Western blot, you can use no HPA-supported tissue or cell line from the supplied record.
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 the samples.
⚠️Feasibility: XRCC3 is intracellular, but the supplied HPA record identifies no positive or negative control sample.
HPA tissue expression evidence for XRCC3
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 XRCC3 Western Blot Tips
Deeper troubleshooting and optimisation questions for XRCC3, answered from its protein features.
Where should the main XRCC3 band appear?
Band shift · The predicted mass is 37.9 kDa. No observed band position is supplied, so use 37.9 kDa as a reference, not an exact expected migration position.
Should XRCC3 isoforms produce multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no isoform-based explanation for multiple bands.
Can XRCC3 acetylation explain a band shift?
PTM · UniProt lists N-acetylmethionine at position 1, using UniProt numbering. This feature alone does not establish a visible shift or explain a difference between apparent and predicted mass.
Does this guide establish induction of XRCC3?
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 XRCC3?
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 M01068-1 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 XRCC3 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 DNA damage increase the XRCC3 band?
Interpretation · XRCC3 participates in DNA repair, and its nuclear foci appear before DNA damage and persist during repair. These features do not establish an increase in total protein abundance after damage. Compare matched samples before interpreting a band change as induction.
UniProt places XRCC3 in the nucleus, cytoplasm, perinuclear region, and mitochondrion. Account for the fraction tested when comparing band intensity; a weak signal in one fraction alone does not establish low total XRCC3 abundance.
XRCC3 interacts with RAD51C and RAD51 and forms a CX3 complex with RAD51C. Those interactions do not establish that a higher band represents the complex. Treat its identity as unresolved without further evidence.
No signal peptide or propeptide is listed, so the supplied features provide no expected cleavage product. They do not identify the cause of a lower band.
UniProt places XRCC3 in the nucleus, cytoplasm, perinuclear region, and mitochondrion. Account for the fraction tested when comparing band intensity; a weak signal in one fraction alone does not establish low total XRCC3 abundance.
XRCC3 interacts with RAD51C and RAD51 and forms a CX3 complex with RAD51C. Those interactions do not establish that a higher band represents the complex. Treat its identity as unresolved without further evidence.
No signal peptide or propeptide is listed, so the supplied features provide no expected cleavage product. They do not identify the cause of a lower band.
Boster reagents
XRCC3 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-XRCC3 Rabbit Monoclonal Antibody
Cat # M01068-1
The catalog reports one anti-XRCC3 rabbit monoclonal antibody for Western blotting, M01068-1. Its reported reactivity is human, mouse, and rat; the supplied WB image shows XRCC3 in HeLa cell lysate. No other sample validation is provided.
Which to pick: M01068-1 is the only listed option. Its WB image uses HeLa cell lysate, so it has direct evidence for that sample context; check the stated reactivity and your sample before use.
Source: BosterBio XRCC3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O43542.
- Human Protein Atlas. XRCC3 tissue expression.
- PMC11485149 — target-verified WB comparison
- PMC3793007 — target-verified WB comparison
- PMC4341824 — target-verified WB comparison