This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
CYFIP1 / Cytoplasmic FMR1-interacting protein 1 · Western blot design guide
Design a Western Blot for CYFIP1
Source-linked CYFIP1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CYFIP1 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
CYFIP1 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 | ~145.2 kDa | |
| Observed band | ~145 kDa | |
| Gel | 5–20% (catalog M04596) | |
| Positive control | Adrenal gland (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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked CYFIP1 Western Blot Protocol Options
The M04596 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human SH-SY5Y, human U20S, human U251, rat brain, rat C6, mouse brain, mouse Neuro-2a (catalog M04596) |
| Gel % | 5–20% (catalog M04596) |
| Load | 30 ug; reducing conditions (catalog M04596) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M04596) |
| Membrane | nitrocellulose membrane (catalog M04596) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M04596) |
| Primary antibody | M04596 · 1:500 (catalog M04596) |
| Primary incubation | overnight at 4°C (catalog M04596) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M04596) |
| Secondary incubation | 1.5 hour at RT (catalog M04596) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M04596) |
| Detection | ECL (catalog M04596) |
Section 2
What Is the Expected CYFIP1 Western Blot Band Size?
CYFIP1 is predicted at 145.2 kDa and observed near 145 kDa; the small difference has no established cause in the supplied evidence.
| Dominant band near 145 kDa | Matches the empirical CYFIP1 band and its 145.2 kDa predicted mass; confirm identity with a control |
| Single sharp band near 145 kDa | Consistent with CYFIP1; the three annotated isoforms need not resolve separately |
| Several discrete bands | Isoforms 1, 2 and 3 are annotated, but their migration and band identities are unverified |
| Band away from 145 kDa | An isoform is possible, but its mass and migration are not supplied; verify identity |
💡Expected CYFIP1 appearanceCYFIP1 is predicted at 145.2 kDa and observed near 145 kDa in reducing whole-cell blots; use band-identity controls to confirm the signal in your sample.
| Predicted CYFIP1 mass | 145.2 kDa predicts a band near the empirical 145 kDa signal |
| Isoform 1 | Annotated splice form; its individual mass and migration are not supplied |
| Isoform 2 | Annotated splice form; its size relative to the other isoforms is unknown |
| Isoform 3 | Annotated splice form; its size relative to the other isoforms is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Insufficient CYFIP1 signal in the cytoplasmic sample | Check sample loading and compare with a positive-control lysate |
| Band higher than expected | Isoform migration or nonspecific binding is possible; neither is established | Compare with the 145 kDa control band and validate identity by CYFIP1 depletion |
| Band lower than expected | An isoform or degradation product is possible; its identity is unverified | Use fresh lysate with protease inhibitors and validate by CYFIP1 depletion |
| Multiple bands | Three splice isoforms are annotated, but distinct bands are unproven | Check which bands diminish after CYFIP1 depletion |
| Weak or no signal | Low sample signal or inefficient transfer of the large protein | Check loading and transfer near 145 kDa against a positive-control lysate |
| Fragments below expected size | Proteolysis during sample preparation is possible | Prepare fresh lysate with protease inhibitors and check fragment identity by CYFIP1 depletion |
Experiment evidence
Western blot images showing CYFIP1 band
M04596 · CYFIP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CYFIP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CYFIP1 in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (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: Whole-cell lysate is suitable for cytoplasmic CYFIP1, though the listed positive tissues show only medium expression.
HPA tissue expression evidence for CYFIP1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | glial 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 → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Low | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced CYFIP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for CYFIP1, answered from its protein features.
How should CYFIP1 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 could CYFIP1 isoforms affect band interpretation?
Isoforms · Three isoforms are listed. Relative to the canonical sequence, isoform 2 lacks residues 1–431 and replaces residues 432–557; isoform 3 lacks residues 1–806. Antibody recognition and apparent size may therefore differ by isoform. Check which sequence your antibody recognizes.
Which CYFIP1 phosphorylation sites are listed?
PTM · UniProt lists phosphoserine at position 583 and phosphothreonine at position 1234. These are canonical UniProt coordinates; antibody or paper numbering may differ. Neither site alone establishes a visible band shift.
Phosphorylation at canonical Ser583 or Thr1234 is a possibility to test, but the listed features do not show that either modification changes gel mobility. Compare matched samples with and without phosphatase treatment before assigning an extra band to phosphorylation.
Phosphorylation at canonical Ser583 or Thr1234 is a possibility to test, but the listed features do not show that either modification changes gel mobility. Compare matched samples with and without phosphatase treatment before assigning an extra band to phosphorylation.
Does this guide establish induction of CYFIP1?
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 CYFIP1 Western blot?
Transfer · CYFIP1 is predicted at 145.2 kDa. Use transfer conditions validated for proteins near 145 kDa, and check transfer with total-protein staining. The supplied features do not specify a membrane or transfer protocol.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M04596 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 CYFIP1 bands be quantified?
Quantitation · Quantify bands consistently within the same apparent-size range and confirm that the signal reflects the isoform your antibody detects. CYFIP1 has three listed isoforms with substantial sequence differences, so combining distinct bands may obscure isoform-specific changes.
Does CYFIP1 show a band shift from its predicted mass?
Interpretation · The reported band is about 145 kDa, close to the predicted 145.2 kDa for canonical CYFIP1. The listed features do not establish a meaningful mass shift.
Consider the listed alternative sequences: isoform 2 lacks canonical residues 1–431, and isoform 3 lacks residues 1–806. A lower band is not sufficient to identify either isoform. Check antibody epitope coverage and whether the band matches the expected isoform-specific sequence.
Consider the listed alternative sequences: isoform 2 lacks canonical residues 1–431, and isoform 3 lacks residues 1–806. A lower band is not sufficient to identify either isoform. Check antibody epitope coverage and whether the band matches the expected isoform-specific sequence.
Boster reagents
CYFIP1 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-CYFIP1 Rabbit Monoclonal Antibody
Cat # M04596
The catalog reports one anti-CYFIP1 Western blot antibody, M04596, with reported human, mouse, and rat reactivity. Its product image shows a band near the expected 145 kDa in the named cell and brain lysates. No independent publication evidence was supplied.
Which to pick: M04596 is the only listed option and has a Western blot image. Its caption documents human cell, rat brain and cell, and mouse brain and cell lysates using a 1:500 primary dilution; use those tested contexts to guide your choice.
Source: BosterBio CYFIP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q7L576.
- Human Protein Atlas. CYFIP1 tissue expression.
- PMC13133249 — target-verified WB comparison
- PMC6555997 — target-verified WB comparison