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- Table of Contents
ACBD6 / Acyl-CoA-binding domain-containing protein 6 · Western blot design guide
Design a Western Blot for ACBD6
Source-linked ACBD6 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ACBD6 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
ACBD6 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 | ~31.2 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Caudate (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 | 1 isoform(s) |
Section 1
Source-Linked ACBD6 Western Blot Protocol Options
The A33955 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 | A33955; 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 ACBD6 Western Blot Band Size?
ACBD6 is predicted at 31.2 kDa; Ser106 phosphorylation is possible, but its migration effect and any empirical band size are unestablished.
| Single band near 31.2 kDa | Consistent with predicted monomeric ACBD6; identity requires controls |
| Band in cytoplasmic fraction | Consistent with cytoplasmic ACBD6 |
| Band in nuclear fraction | Consistent with nuclear ACBD6 |
| No consistent band near twice the monomer size | Consistent with the annotated monomeric state |
💡Expected ACBD6 appearanceACBD6 is predicted at 31.2 kDa, but no empirical band size is supplied; Ser106 phosphorylation has no established migration effect, so confirm a candidate band with identity controls.
| Predicted molecular mass | Places the expected monomer near 31.2 kDa |
| Ser106 phosphorylation | A migration effect is not established |
| Monomeric state | Supports a monomer-size band rather than a dimer-size band |
| Single annotated isoform | Provides no basis for splice-isoform size differences |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ACBD6 may be below detection in the sampled lysate | Check loading and antibody performance with a positive control |
| Band higher than expected | Identity or anomalous migration is unresolved by the supplied features | Confirm with a second antibody or ACBD6 depletion |
| Band lower than expected | Identity or protein degradation is unresolved | Check sample integrity and confirm by ACBD6 depletion |
| Multiple bands | The single annotated isoform does not explain their identities | Identify the ACBD6 band by depletion or a second antibody |
| Weak or no signal | ACBD6 abundance or antibody sensitivity may be limiting | Check loading and use a positive control |
Experiment evidence
Western blot images showing ACBD6 band
A33955 · ACBD6
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ACBD6 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ACBD6 in Western blot, you can use caudate tissue, which shows high expression in HPA.
Positive control: Caudate (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: A tissue negative control is feasible because HPA reports ACBD6 as not detected in adipose tissue.
HPA tissue expression evidence for ACBD6
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 |
|---|---|---|---|---|
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | High | 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 → |
| Cervix | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced ACBD6 Western Blot Tips
Deeper troubleshooting and optimisation questions for ACBD6, answered from its protein features.
Where should the ACBD6 band appear?
Band shift · The predicted mass is 31.2 kDa, but no observed Western blot band is supplied. Use 31.2 kDa as a starting point, not an exact apparent mass; the listed features do not establish a visible shift.
Could ACBD6 isoforms explain multiple bands?
Isoforms · UniProt lists one isoform and no alternative sequence. These features do not support assigning multiple bands to annotated isoforms.
Could phosphorylation affect ACBD6 band interpretation?
PTM · UniProt lists phosphoserine at position 106 in its sequence numbering. A phosphorylation-sensitive assay could help assess its contribution, but the feature alone does not establish a visible band shift.
Does this guide establish induction of ACBD6?
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 ACBD6?
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 A33955 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 ACBD6 be quantified across cellular fractions?
Quantitation · ACBD6 is reported in both cytoplasm and nucleus. Quantify each fraction with appropriate loading controls when comparing localization; a change in one fraction alone need not represent a change in total ACBD6.
Does a higher band represent an ACBD6 oligomer?
Interpretation · UniProt describes ACBD6 as a monomer. That annotation does not support identifying a higher band as its usual oligomeric form. Confirm band identity before assigning it to ACBD6.
No signal peptide or propeptide is listed, so the supplied features do not support a specific processing product. Confirm a lower band's identity before attributing it to ACBD6.
No signal peptide or propeptide is listed, so the supplied features do not support a specific processing product. Confirm a lower band's identity before attributing it to ACBD6.
Boster reagents
ACBD6 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-ACBD6 Antibody
Cat # A33955
The catalog reports one anti-ACBD6 polyclonal antibody for WB, A33955, with stated human, mouse, and rat reactivity. A WB image is listed, but the supplied caption excerpt does not identify the cells tested or establish validation across all three species.
Which to pick: A33955 is the only listed option. Its stated reactivity covers human, mouse, and rat, and it has a WB image. Check the full validation caption for the tested sample and conditions before applying it to your experiment.
Source: BosterBio ACBD6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9BR61.
- Human Protein Atlas. ACBD6 tissue expression.
- PMC3348897 — target-verified WB comparison
- PMC9775454 — target-verified WB comparison
- PMC4398506 — target-verified WB comparison