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- Table of Contents
DNAJB6 / DnaJ homolog subfamily B member 6 · Western blot design guide
Design a Western Blot for DNAJB6
Source-linked DNAJB6 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DNAJB6 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
DNAJB6 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 | ~36.1 kDa | |
| Observed band | ~27 kDa | |
| Gel | 10% (catalog A03290-1) | |
| Positive control | Heart muscle (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 + Methylated | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked DNAJB6 Western Blot Protocol Options
The A03290-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat brain, mouse brain (catalog A03290-1) |
| Gel % | 10% (catalog A03290-1) |
| Load | 30 ug; reducing conditions (catalog A03290-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03290-1) |
| Membrane | nitrocellulose membrane (catalog A03290-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03290-1) |
| Primary antibody | A03290-1 · 0.5 μg/mL (catalog A03290-1) |
| Primary incubation | overnight at 4°C (catalog A03290-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03290-1) |
| Secondary incubation | 1.5 hour at RT (catalog A03290-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03290-1) |
| Detection | ECL (catalog A03290-1) |
Section 2
What Is the Expected DNAJB6 Western Blot Band Size?
UniProt predicts 36.1 kDa; antibody QC reports about 27 kDa in reducing brain lysates, and the cause of this difference is not established.
| Band near 27 kDa | Empirical DNAJB6 band reported in reducing brain lysates; confirm identity with controls. |
| Band near 36 kDa | Near the 36.1 kDa UniProt sequence prediction; identity requires confirmation. |
| Several bands at different positions | Isoforms A, B, C and D are annotated, but distinct band positions are unverified. |
| Higher band after incomplete denaturation | Could reflect retained DNAJB6 homooligomers; its position and identity are unverified. |
💡Expected DNAJB6 appearanceUniProt predicts 36.1 kDa, while antibody QC reports a DNAJB6 band near 27 kDa in reducing brain lysates; the difference is unexplained, so confirm band identity with ordinary controls.
| UniProt predicted mass | 36.1 kDa by sequence; the reported band is near 27 kDa, with no established reason for the difference. |
| Isoform A | Its individual mass and migration are not supplied. |
| Isoform B | Its individual mass and migration are not supplied. |
| Isoform C | Its individual mass and migration are not supplied. |
| Isoform D | Its individual mass and migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| Band lower than expected | The reported 27 kDa band is below the 36.1 kDa prediction; its cause is not established. | Compare with the reported brain lysate result and confirm identity by DNAJB6 depletion or a second antibody. |
| Band higher than expected | DNAJB6 forms homooligomers, which might persist if denaturation is incomplete. | Repeat with thorough denaturation and compare reducing conditions; confirm the band with a second antibody. |
| Multiple bands | Four splice isoforms are annotated, but their band positions are unknown. | Check antibody epitope coverage and compare bands after DNAJB6 depletion. |
| Weak or no signal | DNAJB6 may be unevenly represented across the cytoplasmic, nuclear and muscle Z-line compartments. | Check sample fraction and loading, and compare with a positive lysate control. |
| No band in lysate | The sampled lysate may contain too little detectable DNAJB6. | Verify loading and extraction, then compare with a positive brain lysate control. |
Experiment evidence
Western blot images showing DNAJB6 band
A03290-1 · DNAJB6
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DNAJB6 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DNAJB6 in Western blot, you can use heart muscle lysate, which HPA rates as highly expressed.
Positive control: Heart muscle (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 reports high expression in heart muscle and no detection in adipose tissue.
HPA tissue expression evidence for DNAJB6
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 |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | High | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | High | Protein (IHC) | HPA → |
| Testis | spermatogonia cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | lymphoid tissue | Medium | 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 → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Low | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced DNAJB6 Western Blot Tips
Deeper troubleshooting and optimisation questions for DNAJB6, answered from its protein features.
How should DNAJB6 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.
How can DNAJB6 isoforms affect band interpretation?
Isoforms · UniProt lists isoforms A, B, C and D. B lacks the canonical 242–326 segment; C replaces residues 301–326; D replaces residues 1–59 with a shorter sequence. Check whether the antibody epitope is retained in each isoform before assigning or comparing bands.
For broad isoform detection, choose a region retained across the listed sequence changes and verify the antibody’s stated epitope. Avoid canonical residues 1–59 for isoform D and 232–326 for isoform B. An epitope within 301–326 also encounters a replacement in isoform C.
For broad isoform detection, choose a region retained across the listed sequence changes and verify the antibody’s stated epitope. Avoid canonical residues 1–59 for isoform D and 232–326 for isoform B. An epitope within 301–326 also encounters a replacement in isoform C.
Which DNAJB6 modifications should I consider when interpreting bands?
PTM · The supplied UniProt features list omega-N-methylarginine at canonical residue 135 and phosphoserine at canonical residue 277. Isoform B lacks the segment containing canonical residue 277. These coordinates use canonical UniProt numbering; feature presence alone does not establish a visible shift or explain the ~27 kDa band.
Does this guide establish induction of DNAJB6?
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 DNAJB6?
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 A03290-1 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 DNAJB6 across samples?
Quantitation · Quantify consistently defined bands and record which isoforms the antibody can detect. DNAJB6 is annotated in the nucleus and in cytoplasmic locations, including the perinuclear region and sarcomere Z line, so keep the sampled fraction consistent across comparisons. A change in one band does not by itself measure every isoform.
Why might DNAJB6 appear near 27 kDa instead of 36.1 kDa?
Interpretation · The predicted 36.1 kDa refers to the canonical sequence. Isoform B replaces residues 232–241 and lacks residues 242–326, making it a plausible contributor to a lower band. An observed ~27 kDa band cannot be assigned to isoform B from size alone; confirm which sequence the antibody recognizes.
DNAJB6 is annotated as a homooligomer, which makes oligomers a possibility when investigating higher-mass signal. The annotation does not identify any particular Western-blot band. Compare bands under consistent denaturing preparation and confirm antibody specificity before assigning them.
First compare it with the documented sequence changes: isoform B lacks residues 242–326, while isoform D has a shorter replacement for residues 1–59. Check whether the antibody epitope is present in those isoforms. Apparent mass alone cannot distinguish an isoform from another source of antibody signal.
DNAJB6 is annotated as a homooligomer, which makes oligomers a possibility when investigating higher-mass signal. The annotation does not identify any particular Western-blot band. Compare bands under consistent denaturing preparation and confirm antibody specificity before assigning them.
First compare it with the documented sequence changes: isoform B lacks residues 242–326, while isoform D has a shorter replacement for residues 1–59. Check whether the antibody epitope is present in those isoforms. Apparent mass alone cannot distinguish an isoform from another source of antibody signal.
Boster reagents
DNAJB6 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-DNAJB6 Antibody Picoband®
Cat # A03290-1
The catalog reports one anti-DNAJB6 antibody, A03290-1, with reported Human, Mouse, and Rat reactivity. Its Western blot image shows rat and mouse brain lysates. The caption reports a band near 27 kDa, compared with an expected size of 36 kDa.
Which to pick: A03290-1 is the only listed option. Its WB image uses rat and mouse brain lysates with antibody at 0.5 μg/mL. Human reactivity is listed, but the supplied image does not show a human sample.
Source: BosterBio DNAJB6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O75190.
- Human Protein Atlas. DNAJB6 tissue expression.
- PMC12162027 — target-verified WB comparison
- PMC3660857 — target-verified WB comparison
- PMC5364949 — target-verified WB comparison