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- Table of Contents
ABI2 · Western blot design guide
Design a Western Blot for ABI2
Real validated ABI2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ABI2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Last reviewed: May 2026 · Scientific review: Boster Bio technical team
ABI2 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 | ~55.7 kDa | |
| Observed band | 60 kDa | |
| Gel | 10–12% | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation shift | |
| Regulation | LPS-induced | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated ABI2 Western Blot Protocols
Literature-validated Western blot parameters for ABI2 — gel percentage, transfer, blocking, antibody incubation and detection, extracted from published methods.
| Sample / lysate | human K562, human Hela, human SH-SY5Y |
| Gel % | 5–20% |
| Load | 30 ug |
| Transfer | nitrocellulose membrane, 150 mA, 50–90 min |
| Membrane | nitrocellulose |
| Blocking | 5% non-fat milk / TBS, 1.5 h RT |
| Primary antibody | 0.5 µg/mL |
| Primary incubation | overnight at 4 °C |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 |
| Wash | TBS-0.1% Tween, 3 × 5 min |
| Detection | ECL |
| Exposure / imaging | Tanon 5200 |
| Observed band | 56–70 kDa |
Section 2
What Is the Expected ABI2 Western Blot Band Size?
ABI2 has a 55.7 kDa backbone but typically runs at about 60 kDa, with extra nearby bands most plausibly reflecting phosphorylation and alternative splicing.
| Single band around 60 kDa | full-length ABI2 detected near its expected apparent size |
| Several close bands or a doublet near 60 kDa | alternative ABI2 isoforms and/or different phosphorylation states are being resolved |
| Band slightly above the 55.7 kDa prediction | apparent mobility is shifted modestly upward relative to the calculated backbone mass |
| Clear band in whole-cell lysate | fits a cytoplasmic and nuclear intracellular protein |
💡Expected ABI2 appearanceExpect ABI2 as a mainly intracellular band at about 60 kDa in whole-cell lysate, close to its 55.7 kDa predicted mass, with possible nearby minor phosphoform or isoform bands.
| 55.7 kDa predicted molecular mass | sets the backbone size expected for the full-length protein |
| phosphorylation at Ser40 | can contribute to a small upward mobility shift or band splitting |
| phosphorylation at Ser183 | can contribute to a small upward mobility shift or band splitting |
| phosphorylation at Ser227 | can contribute to a small upward mobility shift or band splitting |
| phosphorylation at Thr361 | can contribute to a small upward mobility shift or band splitting |
| phosphorylation at Ser368 | can contribute to a small upward mobility shift or band splitting |
| isoforms 1, 2, 3 and 4 | can produce multiple ABI2 bands of different relative sizes |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | phosphorylated ABI2 can migrate slightly slower than the 55.7 kDa backbone and appear near 60 kDa | treat the 60 kDa band as expected native ABI2 and compare with phosphatase-treated or differently stimulated samples if needed |
| Band lower than expected | a shorter ABI2 splice isoform may be enriched | check whether the antibody epitope is shared across isoforms and compare patterns across cell types |
| Multiple bands | ABI2 has four splice isoforms and documented phosphorylation sites | expect isoform and phosphoform heterogeneity and confirm specificity by knockdown or an independent antibody |
| Weak or no signal | ABI2 abundance may be low in the tested sample despite being intracellular | use whole-cell lysate from a known positive line or brain-derived sample and increase input or exposure |
| No band in lysate | the sample may express little ABI2 or the antibody may not recognize the present isoform well | verify expression in another positive lysate and confirm that the antibody target region is retained in expressed isoforms |
| Fragments below expected size | lower bands are more consistent with proteolysis because no signal peptide or propeptide cleavage is annotated | improve protease inhibition and sample handling and prioritize the intact 60 kDa band |
Experiment evidence
Western blot images showing ABI2 band
A04302 · ABI2
PA2068 · ABI2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ABI2 Western blot
🧪For positive controls for ABI2 in Western blot, you can use adrenal gland lysate, which is a top HPA positive tissue with high expression.
Positive control: Adrenal gland
Negative control: Adipose tissue
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as stain-free, Ponceau, or REVERT alongside.
⚠️Feasibility: Controls are feasible because ABI2 is an intracellular cytoplasmic/nuclear protein with high expression in selected tissues and HPA not-detected tissues available as negatives.
HPA tissue expression evidence for ABI2
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.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ABI2 Western Blot Tips
Deeper troubleshooting and optimisation questions for ABI2, answered from its protein features.
Why might ABI2 run near 60 kDa?
ABI2 is predicted at 55.7 kDa, but the observed band is about 60 kDa. A modest upward shift is plausible because UniProt lists ABI2 as a phosphoprotein with 5 modified residues. Alternative splicing can also change apparent size, so a band slightly above the predicted mass is consistent with the annotated features.
Could ABI2 isoforms explain multiple bands?
Yes. UniProt lists 4 ABI2 isoforms generated by alternative splicing. If your antibody recognizes a shared region, more than one isoform may be detected, giving closely spaced or distinct bands around the expected size range. Check whether the immunogen maps to all isoforms before interpreting extra bands as nonspecific.
Can phosphorylation alter the ABI2 band pattern?
Possibly. ABI2 is annotated as a phosphoprotein and has 5 modified residues in UniProt. Phosphorylation can broaden, slightly retard, or split the main band. If you see a doublet or diffuse signal near 60 kDa, consider that different phosphorylation states may contribute rather than assuming degradation or off-target binding.
What unexpected ABI2 bands are most plausible?
The most feature-supported explanations are alternative splice isoforms and phosphorylation-dependent mobility differences. ABI2 has 4 isoforms and is a phosphoprotein, so extra bands near the main signal can be biologically real. Very high-mass species are less directly supported by UniProt, even though ABI2 functions in a multiprotein WAVE complex.
Should ABI2 be quantified as one band or several?
If ABI2 appears as a tight doublet or cluster near 60 kDa, quantify consistently based on your biological question. Because UniProt supports both alternative splicing and phosphorylation, summing closely related ABI2 bands may better reflect total ABI2, while separate quantification may be useful if you are tracking modification-dependent shifts.
What transfer method to use for ABI2 Western blot?
ABI2 is a mid-size protein at 55.7 kDa, with an observed band near 60 kDa. Standard wet or semi-dry transfer conditions are generally appropriate for this size range. No signal peptide, propeptide, disulfide bonds, or glycosylation are annotated, so UniProt does not suggest any special transfer adjustment beyond routine optimization.
Does ABI2 localization affect band interpretation?
Yes. UniProt places ABI2 in both cytoplasm and nucleus. If you compare whole-cell with fractionated samples, signal intensity can differ by compartment without implying antibody problems. A band present in cytoplasmic and nuclear fractions can still fit the annotation, provided fraction purity controls support the separation.
Boster reagents
Best ABI2 Western Blot Antibodies
BosterBio's ABI2 antibodies are among the best-performing WB antibodies on the market — well cited, thoroughly validated, and orthogonally cross-validated against negative tissues and complementary methods.
Real WB data
Anti-ABI2 Antibody Picoband®
Cat # A04302
Real WB data
Anti-Abl interactor 2 ABI2 Antibody Picoband®
Cat # PA2068
These recommended anti-ABI2 antibodies represent Boster’s best-performing, well-cited reagents for Western blot, thoroughly validated and orthogonally cross-validated against negative tissues and complementary methods to support confident ABI2 detection.
Which to pick: For ABI2 Western blot, both A04302 and PA2068 include WB validation images. Choose based on your preferred antibody format or host from the product cards, since both are catalogued with direct WB evidence.
Source: BosterBio ABI2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NYB9.
- Human Protein Atlas. ABI2 tissue expression.
- PMC11212232 — Inhibition of ABI2 ubiquitination-dependent degradation suppresses TNBC cell growth via down-regulating PI3K/Akt signaling pathway (Cancer cell international, 2024)
- PMC10103708 — PIM1 phosphorylates ABI2 to enhance actin dynamics and promote tumor invasion (The Journal of cell biology, 2023)