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- Table of Contents
ARF1 / ADP-ribosylation factor 1 · Western blot design guide
Design a Western Blot for ARF1
Real validated ARF1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ARF1 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
ARF1 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 | ~20.7 kDa | |
| Observed band | ~18 kDa | |
| Gel | 12% (catalog M01279-1) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Ovary (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | Band below expected mass | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated ARF1 Western Blot Protocols
The M01279-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human THP-1, human PC-3 (catalog M01279-1) |
| Gel % | 12% (catalog M01279-1) |
| Load | 30 ug; reducing conditions (catalog M01279-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M01279-1) |
| Membrane | nitrocellulose membrane (catalog M01279-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M01279-1) |
| Primary antibody | M01279-1 · 1:500 (catalog M01279-1) |
| Primary incubation | overnight at 4°C (catalog M01279-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:1000 (catalog M01279-1) |
| Secondary incubation | 1.5 hour at RT (catalog M01279-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M01279-1) |
| Detection | ECL (catalog M01279-1) |
Section 2
What Is the Expected ARF1 Western Blot Band Size?
ARF1 is predicted at 20.7 kDa, but antibody blots show approximately 18 kDa; the cause of this difference is not established.
| Band near 18 kDa | Empirical ARF1 band in whole-cell and tissue lysates; confirm identity with controls |
| Band near 20.7 kDa | Close to the predicted mass of full-length ARF1; identity needs confirmation |
| Weak band in a membrane-poor extract | ARF1 can associate with the cytoplasmic side of Golgi membranes |
| Band in whole-cell lysate | ARF1 is detectable in the supplied lysate blots |
💡Expected ARF1 appearanceARF1 has a predicted mass of 20.7 kDa, while antibody blots show a band near 18 kDa; the cause of the difference is unestablished, so confirm identity with ordinary controls.
| Predicted mass of 20.7 kDa | Provides the full-length sequence-based size reference |
| 181-residue sequence | Defines the full-length protein underlying the predicted mass |
| Alternative N-acetylglycine at residue 2 | Has no demonstrated visible migration effect in the supplied evidence |
| Myristate annotation | Lists a lipid modification without establishing an apparent size shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated ARF1 may be poorly recovered | Check extraction and include a positive-control lysate |
| Band higher than expected | Its identity or migration is unestablished | Compare with a positive control and verify antibody specificity |
| Band lower than expected | An approximately 18 kDa band is reported despite the 20.7 kDa prediction | Compare with the reported band and verify identity with a specific control |
| Multiple bands | Additional bands are not explained by the single listed isoform | Check antibody specificity and compare with a positive control |
| Weak or no signal | Low recovery of membrane-associated ARF1 is possible | Check sample extraction, loading and transfer with a positive control |
Experiment evidence
Western blot images showing ARF1 band
M01279-1 · ARF1
M01279-1 · ARF1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ARF1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ARF1 in Western blot, you can use adrenal gland lysate, which HPA rates High.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Ovary (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA reports ARF1 as Not detected in ovary, providing a potential negative tissue control.
HPA tissue expression evidence for ARF1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced ARF1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ARF1, answered from its protein features.
How should ARF1 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 ARF1 isoforms explain multiple bands?
Isoforms · Only one isoform is listed, with no alternative sequence. The supplied features therefore do not support assigning multiple bands to ARF1 isoforms.
Which ARF1 modification should I consider when interpreting bands?
PTM · UniProt lists alternate N-acetylglycine at position 2 and includes myristate among its keywords. Position 2 uses UniProt sequence numbering. These features alone do not establish a visible band shift.
Does this guide establish induction of ARF1?
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 ARF1 Western blot?
Transfer · ARF1 is a small, 181-residue protein with a predicted mass of 20.7 kDa. Check that your transfer conditions retain proteins around 18–21 kDa on the membrane; a 0.2 µm membrane is a practical option.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01279-1 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 I quantify ARF1 across membrane and soluble fractions?
Quantitation · ARF1 is annotated on the cytoplasmic side of the Golgi membrane, with lipid anchoring; its GDP-bound form associates transiently with the membrane. Quantify comparable fractions separately and normalize each to its own loading control. A change in one fraction need not indicate a change in total ARF1.
Why might ARF1 run near 18 kDa instead of 20.7 kDa?
Interpretation · The supplied observed band is about 18 kDa, while the predicted mass is 20.7 kDa. The listed features do not establish why these values differ. Confirm the band with an ARF1-specific control before assigning it.
The supplied features list no glycosylation sites, disulfides, or alternative isoforms, so they do not support those explanations. The listed modification also does not establish a visible shift. Verify unexpected bands with an ARF1-specific control and compare them with the observed band near 18 kDa.
The supplied features list no glycosylation sites, disulfides, or alternative isoforms, so they do not support those explanations. The listed modification also does not establish a visible shift. Verify unexpected bands with an ARF1-specific control and compare them with the observed band near 18 kDa.
Boster reagents
ARF1 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-ARF1 Rabbit Monoclonal Antibody
Cat # M01279-1
The listed anti-ARF1 antibody, M01279-1, has a Western blot image using human, rat, and mouse lysates. The caption reports a band near 18 kDa, compared with an expected 21 kDa; these examples do not establish performance in every sample type.
Which to pick: M01279-1 is the only listed option. Its reported reactivity is human, mouse, and rat, and its WB image includes samples from all three. Check the caption’s sample types and 1:500 primary dilution against your experiment.
Source: BosterBio ARF1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P84077.
- Human Protein Atlas. ARF1 tissue expression.
- PMC12367898 — target-verified WB comparison
- PMC10811507 — target-verified WB comparison
- PMC12278334 — target-verified WB comparison