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- Table of Contents
ARID2 / AT-rich interactive domain-containing protein 2 · Western blot design guide
Design a Western Blot for ARID2
Real validated ARID2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ARID2 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
ARID2 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 | ~197.4 kDa | |
| Observed band | ~245 kDa | |
| Gel | 5–20% (catalog A05064) | |
| 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 C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated ARID2 Western Blot Protocols
The A05064 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human THP-1, human Jurkat, human HEL (catalog A05064) |
| Gel % | 5–20% (catalog A05064) |
| Load | 30 ug; reducing conditions (catalog A05064) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05064) |
| Membrane | nitrocellulose membrane (catalog A05064) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05064) |
| Primary antibody | A05064 · 0.5 μg/mL (catalog A05064) |
| Primary incubation | overnight at 4°C (catalog A05064) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05064) |
| Secondary incubation | 1.5 hour at RT (catalog A05064) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05064) |
| Detection | ECL (catalog A05064) |
Section 2
What Is the Expected ARID2 Western Blot Band Size?
ARID2 is predicted at 197.4 kDa and observed near 245 kDa; the cause of the difference is not established.
| Band near 245 kDa | Empirical ARID2 band reported in reducing whole-cell blots; confirm identity with antibody controls |
| Band near 197.4 kDa | Near the sequence-predicted mass; identity requires confirmation |
| Two bands at different positions | Could reflect isoforms 1 and 2 if their migration differs; distinct bands are not established |
| Weak signal in a cytoplasmic fraction | Consistent with ARID2's nuclear location |
💡Expected ARID2 appearanceARID2 has a predicted mass of 197.4 kDa, while antibody QC blots report a band near 245 kDa; the cause of that difference is unestablished, so confirm identity with antibody and depletion controls.
| Sequence-predicted mass | 197.4 kDa is the calculated reference, while the reported band is near 245 kDa |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its individual mass and migration are not supplied |
| Alternative splicing | Could alter apparent size, but a resolvable isoform pattern is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The reported ARID2 band is near 245 kDa despite a 197.4 kDa predicted mass; the cause is unestablished | Compare with the reported band and confirm identity by ARID2 depletion or immunoprecipitation |
| Band lower than expected | A lower band could represent an isoform or a fragment, but neither assignment is established | Check ARID2 depletion and compare antibodies targeting different regions |
| Multiple bands | ARID2 has isoforms 1 and 2, but their individual migration is unknown | Identify which bands respond to ARID2 depletion |
| Weak or no signal | Nuclear ARID2 may be poorly recovered in the prepared fraction | Check nuclear extraction and loading with a nuclear marker |
| Fragments below expected size | ARID2 fragments have no supplied validated masses | Check sample integrity and confirm candidate bands with ARID2 depletion |
Experiment evidence
Western blot images showing ARID2 band
A05064 · ARID2
A05064 · ARID2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ARID2 Western blot
🧪For positive controls for ARID2 in Western blot, you can use no HPA-supported sample because no positive tissue or cell data were 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 the samples.
⚠️Feasibility: ARID2 is nuclear, but the supplied HPA data cannot establish suitable tissue controls.
HPA tissue expression evidence for ARID2
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 ARID2 Western Blot Tips
Deeper troubleshooting and optimisation questions for ARID2, answered from its protein features.
How should ARID2 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 ARID2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 1784–1835, so it may differ in apparent size or antibody recognition. Check whether the antibody epitope falls in this missing segment when interpreting bands.
Which ARID2 modifications matter when interpreting bands?
PTM · UniProt lists N-acetylalanine at position 2; phosphoserine at 4, 631, 635, 689, 1300, 1391, and 1496; and phosphothreonine at 653 and 692. These are UniProt coordinates and may differ from antibody or paper numbering. Their presence alone does not demonstrate a visible band shift.
Does this guide establish induction of ARID2?
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 ARID2 Western blot?
Transfer · ARID2 has a predicted mass of 197.4 kDa and a supplied observed band near 245 kDa. Use transfer conditions validated for proteins in this size range, and check transfer at the expected band position. The supplied features do not specify a particular membrane or transfer setting.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05064 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 ARID2 bands be quantified?
Quantitation · Keep nuclear sample preparation consistent: UniProt locates ARID2 in the nucleus. Check whether the antibody recognizes the segment missing from isoform 2 (residues 1784–1835), and report which band or bands were quantified.
Why does ARID2 appear near 245 kDa instead of 197.4 kDa?
Interpretation · 197.4 kDa is the predicted mass, while ~245 kDa is the supplied observed band position. The listed modifications and isoforms do not establish the cause of this difference. Check band identity before attributing the higher apparent mass to a specific feature.
Consider the two listed isoforms and whether the antibody epitope lies in residues 1784–1835, which isoform 2 lacks. The listed phosphorylation sites do not, by themselves, identify a shifted band. Verify band identity before assigning an unexpected band to an isoform or modification.
Consider the two listed isoforms and whether the antibody epitope lies in residues 1784–1835, which isoform 2 lacks. The listed phosphorylation sites do not, by themselves, identify a shifted band. Verify band identity before assigning an unexpected band to an isoform or modification.
Boster reagents
ARID2 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-ARID2 Antibody Picoband®
Cat # A05064
The catalog reports one anti-ARID2 antibody, A05064, with Human reactivity and a Western blot image from THP-1, Jurkat, HEL, and MCF-7 whole-cell lysates. The reported band is approximately 245 kDa, versus an expected 197 kDa; the caption does not resolve this difference.
Which to pick: For human ARID2 Western blots, A05064 is the only listed option and has a WB image using four human cell lines. Consider the reported 245 kDa band and expected 197 kDa size when interpreting your result.
Source: BosterBio ARID2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q68CP9.
- Human Protein Atlas. ARID2 tissue expression.
- PMC13224947 — target-verified WB comparison
- PMC7094640 — target-verified WB comparison
- PMC5497798 — target-verified WB comparison