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- Table of Contents
ANKRD1 / Ankyrin repeat domain-containing protein 1 · Western blot design guide
Design a Western Blot for ANKRD1
Real validated ANKRD1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ANKRD1 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
ANKRD1 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 | ~36.3 kDa | |
| Observed band | ~36 kDa | |
| Gel | 5–20% (catalog A04671-2) | |
| Positive control | Heart muscle (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated ANKRD1 Western Blot Protocols
The A04671-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat heart, mouse heart (catalog A04671-2) |
| Gel % | 5–20% (catalog A04671-2) |
| Load | 30 ug; reducing conditions (catalog A04671-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04671-2) |
| Membrane | nitrocellulose membrane (catalog A04671-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04671-2) |
| Primary antibody | A04671-2 · 0.5 μg/mL (catalog A04671-2) |
| Primary incubation | overnight at 4°C (catalog A04671-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04671-2) |
| Secondary incubation | 1.5 hour at RT (catalog A04671-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04671-2) |
| Detection | ECL (catalog A04671-2) |
Section 2
What Is the Expected ANKRD1 Western Blot Band Size?
ANKRD1 is predicted at 36.3 kDa and observed near 36 kDa; the cause of the small difference is not established.
| Band near 36 kDa | Consistent with the empirical ANKRD1 band; confirm identity with controls |
| Single sharp band near 36 kDa | Consistent with the 36.3 kDa predicted mass |
| Little or no band in a cytosolic fraction | Consistent with ANKRD1 nuclear localization |
| Band near 36 kDa in a nuclear fraction | Consistent with ANKRD1 nuclear localization |
💡Expected ANKRD1 appearanceANKRD1 has a predicted mass of 36.3 kDa and an empirical band near 36 kDa; confirm band identity with appropriate controls.
| UniProt predicted mass of 36.3 kDa | Provides the reference for the empirical band near 36 kDa |
| UniProt molecular weight of 36,252 Da | Corresponds to a predicted mass of about 36.3 kDa |
| Predicted mass of full-length ANKRD1 | Places the unprocessed protein near 36.3 kDa |
| Predicted mass of the 319-residue sequence | Places the complete sequence near the observed 36 kDa band |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ANKRD1 may be underrepresented in the sampled fraction | Check nuclear recovery and include a positive control |
| Band higher than expected | No supplied feature establishes a larger ANKRD1 species | Verify band identity with a positive control and antibody specificity control |
| Band lower than expected | No supplied feature establishes a smaller mature product | Check sample integrity and verify band identity |
| Multiple bands | The supplied features do not establish distinct isoform bands | Assess nonspecific binding with an antibody specificity control |
| Weak or no signal | Incomplete recovery of nuclear ANKRD1 may reduce signal | Check nuclear extraction and sample loading |
Experiment evidence
Western blot images showing ANKRD1 band
A04671-2 · ANKRD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ANKRD1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ANKRD1 in Western blot, you can use heart muscle lysate.
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 because HPA reports high expression in heart muscle and no detection in adipose tissue.
HPA tissue expression evidence for ANKRD1
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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ANKRD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ANKRD1, answered from its protein features.
How should ANKRD1 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 annotated isoforms explain multiple ANKRD1 bands?
Isoforms · Only one isoform is listed, with no alternative sequence. The supplied features therefore do not support assigning additional bands to annotated ANKRD1 isoforms.
Do listed modifications predict an ANKRD1 band shift?
PTM · No modified residues or glycosylation sites are listed. Do not assign a shifted band to a specific modification from these features alone.
Does this guide establish induction of ANKRD1?
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 ANKRD1?
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 A04671-2 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 ANKRD1 bands be quantified across samples?
Quantitation · Measure the approximately 36 kDa band consistently across samples. Because ANKRD1 is annotated in the nucleus, keep sample fractionation and loading normalization consistent when comparing signal.
Should ANKRD1 run at its predicted molecular mass?
Interpretation · ANKRD1 has a predicted mass of 36.3 kDa, close to the reported apparent band at approximately 36 kDa. Use that region as a starting point for band identification; the features alone cannot establish the cause of any mobility difference.
ANKRD1 is annotated in the nucleus. Include the nuclear fraction when evaluating detection, and compare the same fraction across samples.
The supplied features identify a 319-residue protein with a predicted mass of 36.3 kDa and one listed isoform. They do not establish the identity of additional bands; verify those bands experimentally before attributing them to ANKRD1.
ANKRD1 is annotated in the nucleus. Include the nuclear fraction when evaluating detection, and compare the same fraction across samples.
The supplied features identify a 319-residue protein with a predicted mass of 36.3 kDa and one listed isoform. They do not establish the identity of additional bands; verify those bands experimentally before attributing them to ANKRD1.
Boster reagents
ANKRD1 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-ANKRD1 Antibody Picoband®
Cat # A04671-2
The catalog reports A04671-2, a rabbit polyclonal anti-ANKRD1 antibody with reported human, mouse, and rat reactivity. Its Western blot image shows rat and mouse heart lysates with a reported band near 36 kDa; no human blot is supplied.
Which to pick: A04671-2 is the only listed option. Its supplied Western blot uses rat and mouse heart lysates at 30 μg per lane and 0.5 μg/mL primary antibody. Human reactivity is listed, but the supplied blot does not test a human sample.
Source: BosterBio ANKRD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15327.
- Human Protein Atlas. ANKRD1 tissue expression.
- PMC4748412 — target-verified WB comparison
- PMC9871652 — target-verified WB comparison
- PMC13333114 — target-verified WB comparison