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- Table of Contents
DNAJC3 / DnaJ homolog subfamily C member 3 · Western blot design guide
Design a Western Blot for DNAJC3
Source-linked DNAJC3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DNAJC3 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
DNAJC3 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 | ~57.6 kDa | |
| Observed band | ~58 kDa | |
| Gel | 5–20% (catalog A04805-1) | |
| Positive control | Bronchus (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 + Cleaved | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked DNAJC3 Western Blot Protocol Options
The A04805-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human U-87MG, human THP-1, human HL-60, rat pancreas, rat testis, mouse pancreas, mouse testis (catalog A04805-1) |
| Gel % | 5–20% (catalog A04805-1) |
| Load | 30 ug; reducing conditions (catalog A04805-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04805-1) |
| Membrane | nitrocellulose membrane (catalog A04805-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04805-1) |
| Primary antibody | A04805-1 · 0.5 μg/mL (catalog A04805-1) |
| Primary incubation | overnight at 4°C (catalog A04805-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04805-1) |
| Secondary incubation | 1.5 hour at RT (catalog A04805-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04805-1) |
| Detection | ECL (catalog A04805-1) |
Section 2
What Is the Expected DNAJC3 Western Blot Band Size?
DNAJC3 is predicted at 57.6 kDa and observed near 58 kDa; the cause of the small difference is not established.
| Single band near 58 kDa | Matches the empirical DNAJC3 band; identity still requires controls |
| Band below 58 kDa | Could reflect removal of the 1–31 signal peptide; migration of the mature form is unreported |
| Two nearby bands | Could reflect precursor and signal-peptide-cleaved forms; distinct migration is unconfirmed |
| No distinct phosphorylation-dependent shift | Phosphoserine 274 does not establish a visibly separate band |
💡Expected DNAJC3 appearanceDNAJC3 has a predicted precursor mass of 57.6 kDa and an empirical band near 58 kDa; confirm identity with ordinary antibody and sample controls.
| Predicted precursor mass | 57.6 kDa; a band near 58 kDa is observed empirically |
| Signal peptide at residues 1–31 | Its presence contributes to the precursor mass |
| Signal-peptide cleavage | Could yield a smaller mature protein; its apparent mass is not supplied |
| Signal-peptide retention | Could preserve precursor size relative to a cleaved form |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ER-localized DNAJC3 may be below detection in the sample | Check an ER-enriched fraction and a positive lysate |
| Band higher than expected | Identity or migration of the higher band is unestablished | Check reduction and confirm with an independent antibody |
| Band lower than expected | Signal-peptide cleavage could reduce size | Compare with the 58 kDa band using an independent antibody |
| Multiple bands | Precursor and cleaved forms are possible, but distinct migration is unconfirmed | Use an independent antibody to assess band identity |
| Weak or no signal | DNAJC3 abundance or detection may be insufficient | Check a positive lysate and antibody conditions |
| Fragments below expected size | Sample proteolysis is possible; no fragment masses are supplied | Repeat with protease inhibitors and confirm with an independent antibody |
Experiment evidence
Western blot images showing DNAJC3 band
A04805-1 · DNAJC3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DNAJC3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DNAJC3 in Western blot, you can use bronchus tissue, which HPA rates High.
Positive control: Bronchus (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: Tissue controls are feasible: HPA rates bronchus High and adipose tissue Not detected.
HPA tissue expression evidence for DNAJC3
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 |
|---|---|---|---|---|
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | 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 → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | Low | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced DNAJC3 Western Blot Tips
Deeper troubleshooting and optimisation questions for DNAJC3, answered from its protein features.
How should DNAJC3 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 isoforms explain multiple DNAJC3 bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore offers no listed isoform explanation for multiple bands. Check whether the bands match the expected ~58 kDa region before assigning them to DNAJC3.
Which DNAJC3 phosphorylation site matters when interpreting bands?
PTM · UniProt lists phosphoserine at position 274, modified by FAM20C. Position 274 uses the supplied UniProt sequence numbering; antibody or paper numbering may differ. The site alone does not establish a visible band shift.
Does this guide establish induction of DNAJC3?
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 DNAJC3?
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 A04805-1 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 DNAJC3 be quantified on a Western blot?
Quantitation · Quantify the consistently identified band near the reported ~58 kDa position across samples. If additional bands appear, do not combine them with that signal solely because DNAJC3 has a signal peptide, phosphoserine 274, or disulfide bonds; the supplied features do not establish those bands’ identities.
Why is the observed DNAJC3 band near 58 kDa?
Interpretation · The reported ~58 kDa band is close to the 57.6 kDa predicted mass. DNAJC3 has a signal peptide at residues 1–31, but these features do not establish how much processing changes its apparent migration. Do not assign a small mass difference to phosphorylation without further evidence.
The UniProt sequence includes a signal peptide at residues 1–31. Consider processing when comparing a detected band with the 57.6 kDa predicted mass of the full sequence; the supplied features do not give a measured mass for the processed protein.
The supplied interaction feature reports DNAJC3 binding to EIF2AK4/GCN2 under ER stress, hypothermic stress, and amino acid starvation, inhibiting its kinase activity. It does not establish that DNAJC3 abundance increases under those conditions; measure abundance directly in an induction experiment.
Compare them with the ~58 kDa reported band. DNAJC3 has a 1–31 signal peptide, phosphoserine 274, and two disulfide bonds, but feature presence alone does not identify an unexpected band or prove altered migration. The supplied record lists no glycosylation sites or alternative sequence.
The UniProt sequence includes a signal peptide at residues 1–31. Consider processing when comparing a detected band with the 57.6 kDa predicted mass of the full sequence; the supplied features do not give a measured mass for the processed protein.
The supplied interaction feature reports DNAJC3 binding to EIF2AK4/GCN2 under ER stress, hypothermic stress, and amino acid starvation, inhibiting its kinase activity. It does not establish that DNAJC3 abundance increases under those conditions; measure abundance directly in an induction experiment.
Compare them with the ~58 kDa reported band. DNAJC3 has a 1–31 signal peptide, phosphoserine 274, and two disulfide bonds, but feature presence alone does not identify an unexpected band or prove altered migration. The supplied record lists no glycosylation sites or alternative sequence.
Boster reagents
DNAJC3 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-DNAJC3 Antibody Picoband®
Cat # A04805-1
A04805-1 is listed for human, mouse, and rat DNAJC3. Its WB image shows an approximately 58 kDa band in three human cell lines and rat and mouse pancreas and testis lysates. No independent publication evidence was supplied.
Which to pick: A04805-1 is the only listed option. Its WB image includes human U-87MG, THP-1, and HL-60 lysates, plus rat and mouse pancreas and testis lysates; use those tested contexts to guide your choice.
Source: BosterBio DNAJC3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13217.
- Human Protein Atlas. DNAJC3 tissue expression.
- PMC11528127 — target-verified WB comparison
- PMC13583059 — target-verified WB comparison
- PMC4778784 — target-verified WB comparison