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- Table of Contents
RAD52 / DNA repair protein RAD52 homolog · Western blot design guide
Design a Western Blot for RAD52
Real validated RAD52 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RAD52 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
RAD52 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 | ~46.2 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 | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated RAD52 Western Blot Protocols
The M01580 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | PC3 cell lysate (catalog M01580) |
| 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 | M01580; 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 RAD52 Western Blot Band Size?
RAD52 has a predicted monomer mass of 46.2 kDa; isoforms and heptameric rings are documented, but their effects on Western blot migration are unproven.
| Band near 46.2 kDa | Consistent with the predicted RAD52 monomer; confirm identity with a control. |
| Additional bands at different positions | Could reflect RAD52 splice isoforms; their migration is not established. |
| Higher molecular weight band | Could reflect incomplete dissociation of RAD52 heptameric rings; verify its identity. |
| Weak or absent band in a cytosolic fraction | Consistent with RAD52 nuclear localization. |
💡Expected RAD52 appearanceThe predicted RAD52 monomer is 46.2 kDa; no empirical band size is supplied, and the documented isoforms and heptameric rings do not establish distinct band positions without validation.
| Predicted RAD52 monomer mass | Provides a 46.2 kDa reference for the full-length protein. |
| Alpha splice isoform | Its specific mass and migration are not supplied. |
| Beta splice isoform | Its specific mass and migration are not supplied. |
| Gamma splice isoform | Its specific mass and migration are not supplied. |
| Delta splice isoform | Its specific mass and migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear RAD52 may be poorly recovered during extraction. | Check nuclear extraction and compare with a nuclear-enriched sample. |
| Band higher than expected | Incomplete dissociation of RAD52 heptameric rings is possible. | Check denaturation conditions and confirm band identity with RAD52 depletion. |
| Band lower than expected | An isoform or degradation product is possible; its mass is not established. | Compare with RAD52 depletion and an antibody against another epitope. |
| Multiple bands | RAD52 has alpha, beta, gamma and delta splice isoforms, but their band positions are unknown. | Use isoform-specific controls or RAD52 depletion to identify the bands. |
| Weak or no signal | RAD52 may be under-recovered when the nuclear fraction is lost. | Check nuclear recovery with a nuclear marker and verify RAD52 antibody performance. |
| Fragments below expected size | Protein degradation during sample preparation is possible. | Prepare fresh lysate with protease inhibitors and confirm fragment identity with RAD52 depletion. |
Experiment evidence
Western blot images showing RAD52 band
M01580 · RAD52
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RAD52 Western blot
🧪For positive controls for RAD52 in Western blot, you can use a RAD52-positive nuclear lysate, although the supplied HPA data identify no specific positive sample.
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 such as Ponceau alongside the samples.
⚠️Feasibility: HPA tissue data are unavailable, so specific tissue controls cannot be selected from the supplied evidence.
HPA tissue expression evidence for RAD52
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 RAD52 Western Blot Tips
Deeper troubleshooting and optimisation questions for RAD52, answered from its protein features.
How should RAD52 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 RAD52 isoforms produce smaller bands?
Isoforms · Yes. UniProt lists alpha, beta, gamma, and delta. Beta lacks canonical residues 227–418; gamma lacks 140–418; delta lacks 119–418. These shorter sequences make lower bands plausible, but band position alone cannot identify an isoform.
Check the antibody epitope against each sequence. An epitope within canonical residues 227–418 is absent from beta, gamma, and delta. Other regions may be replaced: 157–226 in beta, 117–139 in gamma, and 94–118 in delta. An antibody against one of those regions may not recognize every isoform.
Check the antibody epitope against each sequence. An epitope within canonical residues 227–418 is absent from beta, gamma, and delta. Other regions may be replaced: 157–226 in beta, 117–139 in gamma, and 94–118 in delta. An antibody against one of those regions may not recognize every isoform.
Which RAD52 phosphorylation sites should guide band interpretation?
PTM · UniProt lists phosphotyrosine 104 by ABL1, phosphoserine 199, and phosphothreonines 318 and 335, using canonical UniProt coordinates. These sites do not establish a visible shift. Residues 318 and 335 are absent from beta, gamma, and delta; residue 199 is replaced in beta and absent from gamma and delta.
Does this guide establish induction of RAD52?
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 RAD52 Western blot?
Transfer · Plan transfer around canonical RAD52 at 46.2 kDa and the shorter annotated isoforms. Verify transfer across that size range with a total-protein stain. The supplied features do not determine a specific membrane, buffer, or transfer time.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01580 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 RAD52 bands be quantified across samples?
Quantitation · Define whether the measurement is for canonical RAD52 or all detected isoforms, then use the same band boundaries across samples. The four annotated isoforms may yield distinct signals, and antibody epitope location can change which are detected. Report each resolved band separately if isoform identity is uncertain.
How should the predicted mass guide RAD52 band identification?
Interpretation · The canonical RAD52 sequence is 418 residues with a predicted mass of 46.2 kDa. No observed band position is supplied. Use 46.2 kDa as a reference, but do not assign a shifted band to phosphorylation or another cause from feature presence alone.
Compare them with the 46.2 kDa canonical prediction and the shorter beta, gamma, and delta sequences. Check whether the antibody epitope is retained in each isoform. RAD52 also forms heptameric rings, but that feature alone does not identify a high-mass band; likewise, annotated phosphorylation alone does not establish a shifted band.
Compare them with the 46.2 kDa canonical prediction and the shorter beta, gamma, and delta sequences. Check whether the antibody epitope is retained in each isoform. RAD52 also forms heptameric rings, but that feature alone does not identify a high-mass band; likewise, annotated phosphorylation alone does not establish a shifted band.
Boster reagents
RAD52 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-RAD52 Rabbit Monoclonal Antibody
Cat # M01580
M01580 is a rabbit monoclonal anti-RAD52 antibody listed for human reactivity. Its supplied Western blot image shows RAD52 expression in PC3 cell lysate; no other sample validation is provided here.
Which to pick: M01580 is the only listed option. It has a Western blot image using PC3 cell lysate, making it the documented choice for that tested context.
Source: BosterBio RAD52 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P43351.
- Human Protein Atlas. RAD52 tissue expression.
- PMC5650309 — target-verified WB comparison
- PMC6070564 — target-verified WB comparison
- PMC4356517 — target-verified WB comparison