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- Table of Contents
AICDA / Single-stranded DNA cytosine deaminase · Western blot design guide
Design a Western Blot for AICDA
Real validated AICDA Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-AICDA 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
AICDA 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 | ~24 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 + Ubl conjugation | |
| Caveat | Blocking peptide control | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated AICDA Western Blot Protocols
The A00267 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Ramos whole cell lysate (catalog A00267) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Western blot analysis of AID in Ramos whole cell lysate with AID antibody at 2 μg/mL in either the (A) absence or (B) presence of blocking peptide (catalog A00267) |
| Primary antibody | A00267 · 2 μg/mL (catalog A00267) |
| 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 AICDA Western Blot Band Size?
AICDA is predicted at 24 kDa; isoforms and phosphorylation could affect migration, but no empirical band size or visible shift is supplied.
| Band near 24 kDa | Consistent with the predicted AICDA mass; confirm identity with a blocking peptide control. |
| Several bands at different positions | Isoforms 1 and 2 are annotated, but distinct migration has not been demonstrated. |
| Stronger band in cytosolic fraction | Consistent with predominantly cytosolic localization. |
| Weak band in nuclear fraction | Consistent with AICDA being less abundant in the nucleus than in the cytosol. |
💡Expected AICDA appearanceUniProt predicts AICDA at 24 kDa; no empirical band size is supplied, so confirm a band near that position with blocking peptide and sample controls.
| UniProt predicted mass | Full-length AICDA is predicted at 24 kDa. |
| Isoform 1 | May differ in size from isoform 2; its migration is unspecified. |
| Isoform 2 | May differ in size from isoform 1; its migration is unspecified. |
| Phosphothreonine at residue 27 | PKA phosphorylation is annotated; a visible shift is unproven. |
| Phosphoserine at residue 38 | PKA phosphorylation is annotated; a visible shift is unproven. |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | Phosphorylation is annotated, but its effect on migration is unknown; the band may be nonspecific. | Check blocking peptide competition and compare with a phosphatase-treated sample. |
| Band lower than expected | Isoform size differences are possible, but their direction and migration are unspecified. | Check isoform expression and confirm band identity with blocking peptide competition. |
| Broad smear instead of sharp band | The supplied features do not establish a cause for the smear. | Check sample integrity and whether blocking peptide reduces the signal. |
| Multiple bands | Isoforms 1 and 2 are annotated, but the extra bands are unidentified. | Use blocking peptide competition and isoform-specific controls where available. |
| Weak or no signal | AICDA is predominantly cytosolic, so a nuclear fraction may give a weak signal. | Test a cytosolic or whole-cell lysate alongside a positive control. |
Experiment evidence
Western blot images showing AICDA band
A00267 · AICDA
A00267-1 · AICDA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for AICDA Western blot
🧪For positive controls for AICDA in Western blot, you can use no HPA-IHC candidate positive sample from the supplied evidence.
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: HPA provides no tissue candidates, so a positive sample cannot be selected and a knockdown or KO is needed for a negative control.
HPA tissue expression evidence for AICDA
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 AICDA Western Blot Tips
Deeper troubleshooting and optimisation questions for AICDA, answered from its protein features.
What band size should I expect for AICDA?
Band shift · The predicted mass is 24 kDa, but no observed Western blot band size is supplied. Use 24 kDa as a reference, not a confirmed apparent mass.
Could AICDA isoforms produce different bands?
Isoforms · UniProt lists two isoforms. Isoform 2 lacks residues 143–152 relative to isoform 1, so it may migrate differently. An antibody whose epitope includes those residues may fail to detect isoform 2.
Which AICDA phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphothreonine at position 27 and phosphoserine at position 38, both by PKA. These are UniProt coordinates; check the numbering convention before comparing antibody or paper annotations. Their presence does not establish a visible band shift.
Does this guide establish induction of AICDA?
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 AICDA?
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 A00267 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 AICDA 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.
How should I compare AICDA abundance across samples?
Interpretation · AICDA is found in the cytosol and nucleus and is predominantly cytosolic in the supplied annotation. Compare like sample preparations and fractions when quantifying bands, since a change in localization could change the amount recovered from either fraction.
Yes. UniProt places AICDA in both cytosol and nucleus and notes that it is predominantly cytosolic. Record which fraction was loaded and compare matching fractions before interpreting a weaker nuclear or cytosolic signal as a change in total AICDA.
Start with the 24 kDa predicted mass and the two annotated isoforms, including isoform 2’s missing residues 143–152. Consider the annotated PKA phosphorylation sites at UniProt positions 27 and 38, but do not assign a band to phosphorylation from mobility alone. No empirical apparent band size is supplied.
Yes. UniProt places AICDA in both cytosol and nucleus and notes that it is predominantly cytosolic. Record which fraction was loaded and compare matching fractions before interpreting a weaker nuclear or cytosolic signal as a change in total AICDA.
Start with the 24 kDa predicted mass and the two annotated isoforms, including isoform 2’s missing residues 143–152. Consider the annotated PKA phosphorylation sites at UniProt positions 27 and 38, but do not assign a band to phosphorylation from mobility alone. No empirical apparent band size is supplied.
Boster reagents
AICDA 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-AID AICDA Antibody
Cat # A00267
Real WB data
Anti-AID AICDA Antibody
Cat # A00267-1
Both listed anti-AICDA antibodies have Western blot images. A00267 was tested on Ramos whole cell lysate at 2 μg/mL with and without blocking peptide; A00267-1 was shown on HeLa and HepG2 cells. The supplied images do not document every listed reactive species.
Which to pick: For Ramos lysate or a blocking peptide comparison, consider A00267 (listed for human and mouse). For HeLa or HepG2 samples, consider A00267-1 (listed for human, mouse, and rat). Both have WB images; the supplied image evidence does not establish mouse or rat performance.
Source: BosterBio AICDA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9GZX7.
- Human Protein Atlas. AICDA tissue expression.
- PMC3618351 — target-verified WB comparison
- PMC9221089 — target-verified WB comparison
- PMC12171361 — target-verified WB comparison