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- Table of Contents
ARPC2 / Actin-related protein 2/3 complex subunit 2 · Western blot design guide
Design a Western Blot for ARPC2
Source-linked ARPC2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ARPC2 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
ARPC2 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 | ~34.3 kDa | |
| Observed band | ~34 kDa | |
| Gel | 10% (catalog M06165) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked ARPC2 Western Blot Protocol Options
The M06165 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human Hela, human Jurkat (catalog M06165) |
| Gel % | 10% (catalog M06165) |
| Load | 30 ug; reducing conditions (catalog M06165) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M06165) |
| Membrane | nitrocellulose membrane (catalog M06165) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M06165) |
| Primary antibody | M06165 · 1:500 (catalog M06165) |
| Primary incubation | overnight at 4°C (catalog M06165) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M06165) |
| Secondary incubation | 1.5 hour at RT (catalog M06165) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M06165) |
| Detection | ECL (catalog M06165) |
Section 2
What Is the Expected ARPC2 Western Blot Band Size?
ARPC2 is predicted at 34.3 kDa and observed near 34 kDa; the small difference has no established cause in the supplied evidence.
| Single band near 34 kDa | Consistent with ARPC2, subject to band-identity controls |
| Faint band near 34 kDa | May reflect low ARPC2 signal |
| No band near 34 kDa | ARPC2 was not detected in this sample |
| Additional band away from 34 kDa | Identity is uncertain; no distinct ARPC2 band is established by the supplied features |
💡Expected ARPC2 appearanceARPC2 has a predicted mass of 34.3 kDa and an empirical band near 34 kDa; confirm band identity with ordinary controls.
| Predicted molecular mass | Places the expected ARPC2 band near 34.3 kDa |
| Acetylation at Lys275 | A visible migration effect is not established |
| Acetylation at Lys295 | A visible migration effect is not established |
| Arp2/3 complex membership | Does not establish a larger ARPC2 band on a denaturing blot |
| Situation | Likely cause | Next action |
|---|---|---|
| Weak or no signal | ARPC2 may be below the detection limit | Check loading, transfer, antibody performance, and a positive-control lysate |
| Band higher than expected | The supplied features do not establish a higher-migrating ARPC2 species | Compare with a positive control and verify identity by ARPC2 depletion |
| Band lower than expected | The supplied features do not establish a smaller ARPC2 product | Check sample integrity and verify identity by ARPC2 depletion |
| Multiple bands | Only one isoform is listed; additional bands have uncertain identity | Compare with a positive control and verify bands by ARPC2 depletion |
| Fragments below expected size | Sample degradation is possible; no ARPC2 cleavage product is listed | Prepare fresh lysate with protease inhibitors and compare with a positive control |
Experiment evidence
Western blot images showing ARPC2 band
M06165 · ARPC2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ARPC2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ARPC2 in Western blot, you can use appendix tissue lysate, which has high HPA expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: The supplied HPA tissues all show some expression, so use siRNA knockdown or a KO line for a clear negative control.
HPA tissue expression evidence for ARPC2
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 |
|---|---|---|---|---|
| Appendix | lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced ARPC2 Western Blot Tips
Deeper troubleshooting and optimisation questions for ARPC2, answered from its protein features.
How should ARPC2 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.
Do listed ARPC2 isoforms explain multiple bands?
Isoforms · Only one isoform is listed, with no alternative sequence supplied. The supplied features therefore do not support assigning additional bands to specific ARPC2 isoforms.
Could acetylation shift the ARPC2 band?
PTM · UniProt lists N6-acetyllysine at positions 275 and 295, using UniProt sequence numbering. These features alone do not establish a visible band shift. Check an observed shift independently before attributing it to acetylation.
No glycosylation sites are listed, and the glycosylation count is zero. The supplied features do not support glycosylation as an explanation for a heavier band.
No glycosylation sites are listed, and the glycosylation count is zero. The supplied features do not support glycosylation as an explanation for a heavier band.
Does this guide establish induction of ARPC2?
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 ARPC2?
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 M06165 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 ARPC2 be quantified across fractions?
Quantitation · Because ARPC2 is listed in several cellular locations, compare like fractions when assessing changes in its band intensity. A difference between fractions may reflect distribution as well as total ARPC2 abundance.
How should the ARPC2 band compare with its predicted mass?
Interpretation · ARPC2 has a predicted mass of 34.3 kDa, close to the reported apparent band near 34 kDa. A small difference between calculated and apparent mass does not, by itself, establish a modification.
UniProt places ARPC2 in the cytoplasm and cytoskeleton, cell projections, synaptosomes, and nucleus. Consider which fraction was loaded when interpreting a weak or missing band.
Start with the expected band near 34 kDa. ARPC2 is a component of the Arp2/3 complex, but that association alone does not identify another band. The listed acetylation sites and single isoform also do not establish the identity of an unexpected band.
UniProt places ARPC2 in the cytoplasm and cytoskeleton, cell projections, synaptosomes, and nucleus. Consider which fraction was loaded when interpreting a weak or missing band.
Start with the expected band near 34 kDa. ARPC2 is a component of the Arp2/3 complex, but that association alone does not identify another band. The listed acetylation sites and single isoform also do not establish the identity of an unexpected band.
Boster reagents
ARPC2 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-ARPC2 Rabbit Monoclonal Antibody
Cat # M06165
The catalog reports one anti-ARPC2 WB antibody, rabbit monoclonal M06165, with reported human, mouse, and rat reactivity. Its WB image shows a band near the expected 34 kDa in the captioned lysates. No publication evidence is supplied.
Which to pick: M06165 is the only listed option and has a WB image. The caption documents human cell lines, rat skeletal muscle and PC-12, and mouse skeletal muscle and NIH/3T3, using 30 µg reduced lysate and a 1:500 primary antibody dilution.
Source: BosterBio ARPC2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O15144.
- Human Protein Atlas. ARPC2 tissue expression.
- PMC9207441 — target-verified WB comparison
- PMC13027156 — target-verified WB comparison
- PMC3821612 — target-verified WB comparison