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- Table of Contents
WIPI1 / WD repeat domain phosphoinositide-interacting protein 1 · Western blot design guide
Design a Western Blot for WIPI1
Source-linked WIPI1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-WIPI1 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
WIPI1 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 | ~48.7 kDa | |
| Observed band | ~49 kDa | |
| Gel | 10% (catalog M06206-2) | |
| Positive control | Placenta (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 | — | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked WIPI1 Western Blot Protocol Options
The M06206-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HEL, human HepG2, human MCF-7 (catalog M06206-2) |
| Gel % | 10% (catalog M06206-2) |
| Load | 30 ug; reducing conditions (catalog M06206-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M06206-2) |
| Membrane | nitrocellulose membrane (catalog M06206-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M06206-2) |
| Primary antibody | M06206-2 · 1:500 (catalog M06206-2) |
| Primary incubation | overnight at 4°C (catalog M06206-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M06206-2) |
| Secondary incubation | 1.5 hour at RT (catalog M06206-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M06206-2) |
| Detection | ECL (catalog M06206-2) |
Section 2
What Is the Expected WIPI1 Western Blot Band Size?
WIPI1 is predicted at 48.7 kDa and observed at ~49 kDa; the cause of the small difference is not established.
| Band at ~49 kDa | Empirical WIPI1 band in reducing whole-cell lysates; confirm identity with antibody controls |
| Single band near 48.7 kDa | Consistent with the UniProt predicted mass |
| Multiple bands near the expected size | Isoforms 1 and 2 could contribute, but distinct migration is unproven |
| No separate isoform band | Isoforms 1 and 2 are listed, but their separation on a blot is unknown |
💡Expected WIPI1 appearanceWIPI1 has a predicted mass of 48.7 kDa and an empirical band at ~49 kDa in reducing whole-cell lysates; confirm band identity with ordinary antibody controls.
| UniProt predicted mass | Predicts 48.7 kDa; the observed whole-cell band is ~49 kDa |
| Splice isoform 1 | May migrate differently from isoform 2; its individual mass is unspecified |
| Splice isoform 2 | May migrate differently from isoform 1; its individual mass is unspecified |
| Alternative splicing of isoforms 1 and 2 | Could produce more than one band, but a resolvable size difference is unproven |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated WIPI1 may be poorly recovered | Check lysate preparation and include a positive whole-cell lysate |
| Band higher than expected | Its identity and cause of migration are unestablished | Compare with the ~49 kDa control band and check antibody specificity |
| Band lower than expected | An isoform is possible, but its migration is unknown | Compare with the ~49 kDa band and confirm identity with an independent antibody |
| Multiple bands | Isoforms 1 and 2 are annotated, but separate bands are unproven | Check antibody specificity and compare with a positive control |
| Weak or no signal | Recovery of peripheral membrane-associated WIPI1 may be limited | Check extraction and loading with a positive whole-cell lysate |
Experiment evidence
Western blot images showing WIPI1 band
M06206-2 · WIPI1
A06206 · WIPI1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for WIPI1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for WIPI1 in Western blot, you can use placenta lysate, as HPA reports high expression in placenta.
Positive control: Placenta (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: WIPI1 is membrane-associated, so ensure the lysate retains membrane-associated proteins.
HPA tissue expression evidence for WIPI1
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 |
|---|---|---|---|---|
| Placenta | decidual cells | High | Protein (IHC) | HPA → |
| Skin | fibrohistiocytic cells | High | 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced WIPI1 Western Blot Tips
Deeper troubleshooting and optimisation questions for WIPI1, answered from its protein features.
How should WIPI1 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.
Can WIPI1 isoforms 1 and 2 be distinguished by Western blot?
Isoforms · Isoform 2 replaces the first six residues of isoform 1, MEAEAA, with a single methionine. That five-residue difference may be difficult to resolve on a standard gel. A band near 49 kDa alone cannot establish the isoform.
Should a modification cause a visible WIPI1 band shift?
PTM · The supplied features list no modified residues or glycosylation sites. They provide no basis to predict a modification-driven shift; an unexpected band needs independent investigation.
Can autophagy induction change WIPI1 Western blot signal?
Induction · WIPI1 is associated with autophagy and preautophagosomal membranes. Compare matched total lysates when assessing abundance, and use consistent fractionation if measuring membrane-associated WIPI1. These features do not establish that autophagy induction increases WIPI1 expression.
How should transfer be checked for WIPI1?
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 M06206-2 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 WIPI1 be quantified across conditions?
Quantitation · Quantify bands from the same type of preparation, with matched loading and signals within the assay’s linear range. Because WIPI1 occupies cytoplasmic and membrane-associated locations, changes in one fraction alone do not establish a change in total abundance.
Why is the WIPI1 band near 49 kDa?
Interpretation · The observed band near 49 kDa agrees with the predicted 48.7 kDa mass. This agreement does not identify which isoform is present.
WIPI1 is annotated as a peripheral membrane protein at several sites, including preautophagosomal membranes, endosomes, and the trans-Golgi network. Keep extraction and fractionation consistent across samples; a change in the fraction collected could change the measured signal.
Compare it with the expected band near 49 kDa and verify its identity independently. The two annotated isoforms differ by only five residues at the N terminus, and the supplied features do not establish a visible shift from a modification. Band position alone cannot assign an unexpected band to an isoform or modification.
WIPI1 is annotated as a peripheral membrane protein at several sites, including preautophagosomal membranes, endosomes, and the trans-Golgi network. Keep extraction and fractionation consistent across samples; a change in the fraction collected could change the measured signal.
Compare it with the expected band near 49 kDa and verify its identity independently. The two annotated isoforms differ by only five residues at the N terminus, and the supplied features do not establish a visible shift from a modification. Band position alone cannot assign an unexpected band to an isoform or modification.
Boster reagents
WIPI1 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-WIPI1 Rabbit Monoclonal Antibody
Cat # M06206-2
Real WB data
Anti-WIPI1 Antibody
Cat # A06206
The catalog reports two anti-WIPI1 antibodies with WB images. Both list Human, Mouse, and Rat reactivity. M06206-2 shows a band near 49 kDa in three human cell lysates; A06206 is shown with rat colon tissue lysate. No mouse WB example is supplied.
Which to pick: For human cell lysates, start with M06206-2; its image uses HEL, HepG2, and MCF-7 lysates at 1:500. For rat colon tissue, A06206 has a WB example at 1 and 2 μg/mL. Both list mouse reactivity, but neither caption shows a mouse sample.
Source: BosterBio WIPI1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q5MNZ9.
- Human Protein Atlas. WIPI1 tissue expression.
- PMC8980855 — target-verified WB comparison
- PMC12466441 — target-verified WB comparison
- PMC8632285 — target-verified WB comparison