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- Table of Contents
PRMT3 / Protein arginine N-methyltransferase 3 · Western blot design guide
Design a Western Blot for PRMT3
Source-linked PRMT3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PRMT3 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
PRMT3 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 | ~59.9 kDa | |
| Observed band | ~60–70 kDa | |
| Gel | 5–20% (catalog A05694-1) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Liver (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked PRMT3 Western Blot Protocol Options
The A05694-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human MCF-7, human HepG2, human Hela, human 293T, human K562, human Daudi, human HL-60, human MOLT-4 (catalog A05694-1) |
| Gel % | 5–20% (catalog A05694-1) |
| Load | 30 ug; reducing conditions (catalog A05694-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05694-1) |
| Membrane | nitrocellulose membrane (catalog A05694-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05694-1) |
| Primary antibody | A05694-1 · 0.5 μg/mL (catalog A05694-1) |
| Primary incubation | overnight at 4°C (catalog A05694-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05694-1) |
| Secondary incubation | 1.5 hour at RT (catalog A05694-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05694-1) |
| Detection | ECL (catalog A05694-1) |
Section 2
What Is the Expected PRMT3 Western Blot Band Size?
PRMT3 is predicted at 59.9 kDa and observed at ~60–70 kDa; the cause of any difference is not established.
| Band at ~60–70 kDa | Empirical PRMT3 band in reducing whole-cell lysates; confirm identity with antibody controls |
| Band near 59.9 kDa | Consistent with the predicted PRMT3 monomer mass |
| Several nearby bands | Isoforms 1 and 2 may contribute, but distinct migration is unverified |
| Band near twice the monomer size | Could reflect a homodimer if it survives sample preparation |
💡Expected PRMT3 appearancePRMT3 has a predicted mass of 59.9 kDa and an empirical band at ~60–70 kDa in reducing whole-cell lysates; confirm band identity with appropriate controls.
| Predicted PRMT3 monomer mass | 59.9 kDa provides the sequence-based reference |
| Isoform 1 | May migrate differently from isoform 2; its mass is unspecified |
| Isoform 2 | May migrate differently from isoform 1; its mass is unspecified |
| Monomer and homodimer | A retained homodimer could migrate near twice the monomer size; survival through SDS-PAGE is unverified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PRMT3 signal may be below detection in the sampled lysate | Check a positive-control whole-cell lysate and increase loading or exposure |
| Band higher than expected | A homodimer could persist through sample preparation; band identity remains uncertain | Compare reducing and nonreducing samples and verify specificity |
| Band lower than expected | An alternative isoform or nonspecific band is possible; isoform masses are unspecified | Compare antibodies targeting different PRMT3 regions or deplete PRMT3 |
| Multiple bands | Isoforms 1 and 2 are annotated, though distinct migration is unverified | Use PRMT3 depletion to identify which bands are specific |
| Weak or no signal | PRMT3 may be insufficiently represented in the sampled cytosolic or nuclear material | Check a positive-control lysate and assess the relevant fractions |
Experiment evidence
Western blot images showing PRMT3 band
A05694-1 · PRMT3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PRMT3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PRMT3 in Western blot, you can use adipose tissue, which HPA scores as Medium.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Liver (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: PRMT3 is intracellular, and HPA reports liver as not detected, making it a plausible negative tissue control.
HPA tissue expression evidence for PRMT3
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Low | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced PRMT3 Western Blot Tips
Deeper troubleshooting and optimisation questions for PRMT3, answered from its protein features.
How should PRMT3 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 PRMT3 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, the sequence corresponding to canonical residues 11–99 is replaced by a shorter sequence. An antibody's recognition of each isoform depends on its epitope; use an epitope outside that changed region if both isoforms need detection.
Do not silently carry over canonical coordinates. Isoform 2 replaces the region corresponding to canonical residues 11–99, which contains phosphoserines 25 and 27. Map sites to the isoform 2 sequence before interpreting a site-specific signal; UniProt also lists canonical phosphoserine 171.
Do not silently carry over canonical coordinates. Isoform 2 replaces the region corresponding to canonical residues 11–99, which contains phosphoserines 25 and 27. Map sites to the isoform 2 sequence before interpreting a site-specific signal; UniProt also lists canonical phosphoserine 171.
Which PRMT3 modifications matter when assessing band mobility?
PTM · UniProt lists N-acetylcysteine at position 2 and phosphoserine at positions 25, 27, and 171. These are canonical-sequence coordinates. Their presence does not establish that a visible shift will occur or explain any measured mass difference.
Does this guide establish induction of PRMT3?
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 PRMT3?
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 A05694-1 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 PRMT3 band intensity be quantified?
Quantitation · Specify whether the measurement is from whole-cell, cytosolic, or nuclear material, because PRMT3 is reported in both compartments. Quantify the same defined band or bands across samples, accounting for the two listed isoforms if the antibody detects both.
Why might PRMT3 appear at 60–70 kDa?
Interpretation · PRMT3 has a predicted mass of 59.9 kDa, close to the reported apparent band range. UniProt lists two isoforms and several modified residues, but those features alone do not establish why a particular band migrates above 59.9 kDa.
PRMT3 is reported in the cytosol and nucleus, and ZNF200 localizes it to the nucleus. Check both fractions when assessing distribution. A change in one fraction alone need not represent a change in total cellular PRMT3.
Consider the two isoforms and the listed modifications, but do not assign a band to either without supporting evidence. UniProt describes PRMT3 as a monomer and homodimer by similarity; that annotation alone does not identify a Western-blot band. No glycosylation sites, signal peptide, or propeptide are listed.
PRMT3 is reported in the cytosol and nucleus, and ZNF200 localizes it to the nucleus. Check both fractions when assessing distribution. A change in one fraction alone need not represent a change in total cellular PRMT3.
Consider the two isoforms and the listed modifications, but do not assign a band to either without supporting evidence. UniProt describes PRMT3 as a monomer and homodimer by similarity; that annotation alone does not identify a Western-blot band. No glycosylation sites, signal peptide, or propeptide are listed.
Boster reagents
PRMT3 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-PRMT3 Antibody Picoband®
Cat # A05694-1
The listed anti-PRMT3 antibody A05694-1 is a rabbit polyclonal with reported human reactivity. Its Western blot caption reports a 60–70 kDa band across eight human cell lysates; the expected size is 60 kDa. Evidence is limited to the reported conditions.
Which to pick: A05694-1 is the only listed option and has a Western blot image. For human cell lysates, its reported conditions used 30 µg of sample per lane and 0.5 µg/mL primary antibody.
Source: BosterBio PRMT3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O60678.
- Human Protein Atlas. PRMT3 tissue expression.
- PMC10782741 — target-verified WB comparison
- PMC10754120 — target-verified WB comparison