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- Table of Contents
DNAJA1 / DnaJ homolog subfamily A member 1 · Western blot design guide
Design a Western Blot for DNAJA1
Real validated DNAJA1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DNAJA1 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
DNAJA1 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 | ~44.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated DNAJA1 Western Blot Protocols
The M04151 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysate (catalog M04151) |
| 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 | M04151; 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 DNAJA1 Western Blot Band Size?
DNAJA1 has a predicted precursor mass of 44.9 kDa; propeptide cleavage and isoforms could affect size, but their migration and an empirical band are unreported.
| Band near 44.9 kDa | Consistent with the predicted DNAJA1 precursor; confirm its identity with a specific control. |
| Band slightly below 44.9 kDa | Could reflect removal of the three-residue propeptide at 395–397. |
| Bands at different sizes | Could reflect isoforms 1 and 2; distinct migration has not been established. |
| Weak band in a soluble fraction | Could reflect DNAJA1 association with membranes through a lipid anchor. |
💡Expected DNAJA1 appearanceThe predicted precursor mass is 44.9 kDa; propeptide removal and isoforms could affect migration, but no empirical band size or resolvable isoform pattern is supplied, so verify band identity with a specific control.
| Predicted precursor mass | Sets a reference near 44.9 kDa, not a measured Western blot position. |
| Propeptide at residues 395–397 | Its removal could make mature DNAJA1 slightly smaller than the precursor. |
| Splice isoform 1 | Its size relative to isoform 2 is not supplied. |
| Splice isoform 2 | It could migrate differently from isoform 1, but the direction and separation are unknown. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated DNAJA1 may be poorly extracted. | Check extraction and compare soluble and membrane fractions. |
| Band higher than expected | The band may represent modified DNAJA1 or an unrelated protein; a visible shift is unproven. | Check identity with DNAJA1 knockdown and a second antibody. |
| Band lower than expected | Propeptide removal or an isoform is possible; neither has a supplied band size. | Check band identity with DNAJA1 knockdown. |
| Multiple bands | Isoforms 1 and 2 are reported, but their separation on a blot is unknown. | Check which bands decrease after DNAJA1 knockdown. |
| Weak or no signal | DNAJA1 may be underrepresented after fractionation because it associates with membranes. | Compare membrane and soluble fractions and check loading. |
Experiment evidence
Western blot images showing DNAJA1 band
M04151 · DNAJA1
PA1597 · DNAJA1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DNAJA1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DNAJA1 in Western blot, you can use appendix tissue, which HPA scores as high expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA identifies a not-detected tissue for a negative control; use consistent lysate preparation because DNAJA1 occupies multiple intracellular locations.
HPA tissue expression evidence for DNAJA1
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial 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 → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced DNAJA1 Western Blot Tips
Deeper troubleshooting and optimisation questions for DNAJA1, answered from its protein features.
How should DNAJA1 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 DNAJA1 isoforms produce different bands?
Isoforms · Two isoforms are listed. In isoform 2, canonical residues 327..397 are replaced by SCNVL, making it shorter. Check whether the antibody epitope is retained before assigning bands to either isoform. The features do not establish their observed migration.
Which DNAJA1 modifications matter when interpreting bands?
PTM · Canonical UniProt positions list N6-acetyllysine 66, phosphoserine 83 and 335, phosphotyrosine 381, and cysteine methyl ester 394. Isoform 2 replaces the region containing positions 335, 381, and 394. These coordinates use canonical UniProt numbering; antibody or paper numbering may differ. Modification sites alone do not establish visible shifts.
Does this guide establish induction of DNAJA1?
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 DNAJA1 Western blot?
Transfer · The canonical protein is predicted at 44.9 kDa, while isoform 2 has a shorter alternative sequence. Choose transfer conditions suitable for the sizes you intend to detect and verify transfer with a total-protein stain. The supplied features do not specify a transfer method or duration.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M04151 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 DNAJA1 be quantified across cell fractions?
Quantitation · DNAJA1 is annotated in the cytoplasm, nucleus, membranes, and other compartments. Compare bands within consistently prepared fractions, and account for the amount of each fraction loaded. A change in one fraction may reflect redistribution rather than a change in total DNAJA1.
Where should canonical DNAJA1 migrate on a Western blot?
Interpretation · The predicted mass is 44.9 kDa, so examine the region around 45 kDa. No observed band position is supplied. The listed features alone cannot establish an apparent mass or explain a difference from 44.9 kDa.
Check antibody recognition of both isoforms: isoform 2 replaces canonical residues 327..397 with SCNVL. A propeptide is annotated at canonical positions 395..397, near the modified cysteine at 394. These features suggest regions to check when assessing antibody binding, but they do not identify an unexpected band or prove a visible shift.
Check antibody recognition of both isoforms: isoform 2 replaces canonical residues 327..397 with SCNVL. A propeptide is annotated at canonical positions 395..397, near the modified cysteine at 394. These features suggest regions to check when assessing antibody binding, but they do not identify an unexpected band or prove a visible shift.
Boster reagents
DNAJA1 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-DNAJA1 Rabbit Monoclonal Antibody
Cat # M04151
Real WB data
Anti-HDJ2/DNAJA1 Antibody Picoband®
Cat # PA1597
Both catalogued anti-DNAJA1 antibodies list human, mouse, and rat reactivity and have Western blot images. M04151 is shown with Jurkat lysate; PA1597 is shown with rat and mouse tissues and several cell lines. No publication evidence is supplied.
Which to pick: Choose M04151 if the shown Jurkat lysate matches your context. Choose PA1597 if its documented rat or mouse brain and lung tissues, or listed cell-line lysates, better match your sample. Both list the same species reactivity; the images show specific tested samples.
Source: BosterBio DNAJA1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P31689.
- Human Protein Atlas. DNAJA1 tissue expression.
- PMC10995657 — target-verified WB comparison
- PMC6800788 — target-verified WB comparison
- PMC7948449 — target-verified WB comparison