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- Table of Contents
ABI3 / ABI gene family member 3 · Western blot design guide
Design a Western Blot for ABI3
Source-linked ABI3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ABI3 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
ABI3 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 | ~39 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Lymph node (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked ABI3 Western Blot Protocol Options
The A03909 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | A03909; 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 ABI3 Western Blot Band Size?
ABI3 has a predicted mass of 39 kDa; splice isoforms and phosphorylation could affect migration, but no empirical band or visible effect is established.
| Band near 39 kDa | Consistent with the predicted ABI3 mass; identity requires controls |
| Band at a different size | Could reflect an ABI3 splice isoform; its mass is not supplied |
| Multiple bands | Could reflect isoforms 1 and 2 if their migration differs enough to resolve |
| Closely spaced bands | Could reflect different phosphorylation states, but a visible shift is not established |
💡Expected ABI3 appearanceUniProt predicts ABI3 at 39 kDa, but no empirical band size is supplied; use antibody specificity and ABI3 depletion controls to establish band identity.
| UniProt predicted mass | Places the reference band near 39 kDa |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | May differ in size from isoform 1; its individual mass is not supplied |
| Alternative splicing | Could produce distinct bands if isoforms 1 and 2 resolve |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ABI3 abundance in the tested lysate is unknown | Check expression and enrich the cytoplasmic fraction |
| Band higher than expected | An isoform or phosphorylation could affect migration, but the cause is unestablished | Compare with ABI3 depletion and phosphatase-treated samples |
| Band lower than expected | An isoform may differ in size; its mass is unknown | Confirm identity by ABI3 depletion and an antibody to another epitope |
| Multiple bands | Isoforms 1 and 2 or phosphorylation states may differ in migration | Compare isoform expression and phosphatase-treated samples with ABI3 depletion |
| Weak or no signal | ABI3 expression in the tested sample may be low | Check expression and use a known ABI3-positive lysate |
Experiment evidence
Western blot images showing ABI3 band
A03909 · ABI3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ABI3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ABI3 in Western blot, you can use lymph node lysate.
Positive control: Lymph node (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Cytoplasmic ABI3 should be accessible in tissue lysates, and HPA identifies adipose tissue as a negative control.
HPA tissue expression evidence for ABI3
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 |
|---|---|---|---|---|
| Lymph node | non-germinal center cells | High | Protein (IHC) | HPA → |
| Spleen | cells in white pulp | High | Protein (IHC) | HPA → |
| Tonsil | non-germinal center cells | High | Protein (IHC) | HPA → |
| Appendix | non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | endocrine cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ABI3 Western Blot Tips
Deeper troubleshooting and optimisation questions for ABI3, answered from its protein features.
How should ABI3 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 ABI3 isoforms produce nearby bands?
Isoforms · UniProt lists isoforms 1 and 2; isoform 2 lacks residues 90–95. This small sequence difference could affect band position, but the features do not establish whether the isoforms resolve on a blot.
Which ABI3 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserines at positions 213, 216, and 342. These are UniProt coordinates; paper or antibody numbering may differ. Their presence does not establish a visible band shift.
Quantify total ABI3 signal separately from any site-specific phosphoserine signal. For a site-specific measurement, confirm that the antibody targets UniProt position 213, 216, or 342 under the numbering convention used by its supplier.
Quantify total ABI3 signal separately from any site-specific phosphoserine signal. For a site-specific measurement, confirm that the antibody targets UniProt position 213, 216, or 342 under the numbering convention used by its supplier.
Does this guide establish induction of ABI3?
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 ABI3?
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 A03909 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 ABI3 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 ABI3 migrate exactly at its predicted 39 kDa?
Interpretation · The predicted mass is 39 kDa, but no observed band position is supplied. Use 39 kDa as a reference, not a guaranteed migration position. The listed phosphorylation sites and isoforms alone do not establish a visible shift or explain any mass difference.
Compare bands with the 39 kDa prediction and consider the six-residue deletion in isoform 2. Do not assign an unexpected band to an isoform or phosphorylation solely from its position; the supplied features give no observed band pattern.
ABI3 is annotated as cytoplasmic and colocalizes with PAK2 at the leading edge of cells. A cytoplasmic sample is therefore relevant when checking ABI3 detection; localization alone does not specify its abundance in a particular preparation.
ABI3 may interact with PAK1 and PAK2 and probably interacts with TARSH. These annotations do not establish that an additional band contains ABI3 or an interacting protein. Verify band identity independently before interpreting it as a complex or partner.
Compare bands with the 39 kDa prediction and consider the six-residue deletion in isoform 2. Do not assign an unexpected band to an isoform or phosphorylation solely from its position; the supplied features give no observed band pattern.
ABI3 is annotated as cytoplasmic and colocalizes with PAK2 at the leading edge of cells. A cytoplasmic sample is therefore relevant when checking ABI3 detection; localization alone does not specify its abundance in a particular preparation.
ABI3 may interact with PAK1 and PAK2 and probably interacts with TARSH. These annotations do not establish that an additional band contains ABI3 or an interacting protein. Verify band identity independently before interpreting it as a complex or partner.
Boster reagents
ABI3 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-ABI gene family member 3 ABI3 Antibody
Cat # A03909
A03909 is an anti-ABI3 antibody listed as reactive with human and mouse. Its Western blot image uses PC12 cell lysate with primary antibody at 1:1000, incubated overnight at 4°C. The supplied evidence shows this tested sample and condition only.
Which to pick: A03909 is the only listed option. It has a Western blot image from PC12 lysate and is listed for human and mouse reactivity; choose it with your sample context in mind.
Source: BosterBio ABI3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9P2A4.
- Human Protein Atlas. ABI3 tissue expression.
- PMC10013321 — target-verified WB comparison
- PMC9327202 — target-verified WB comparison