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- Table of Contents
CAGE1 / Cancer-associated gene 1 protein · Western blot design guide
Design a Western Blot for CAGE1
Source-linked CAGE1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CAGE1 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
CAGE1 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 | ~90.3 kDa | |
| Gel | 8–10% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Isoform controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked CAGE1 Western Blot Protocol Options
The A12728 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Hela (40ug), HEK293T (40ug), The testis of Mouse(40ug), The testis of Rat(40ug), H9C2 (40ug) (catalog A12728) |
| Gel % | 8–10% (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 | A12728; 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 CAGE1 Western Blot Band Size?
CAGE1 is predicted at 90.3 kDa; five isoforms could affect band patterns, but no distinct migration or empirical band size is established.
| Band near 90.3 kDa | consistent with the predicted CAGE1 mass; identity requires controls |
| Single band near 90.3 kDa | the five annotated isoforms need not resolve into separate bands |
| Additional band at a different size | an isoform is possible, but its migration is unknown |
| Several bands at different sizes | alternative splicing is a possible explanation; confirm band identities |
💡Expected CAGE1 appearanceUniProt predicts CAGE1 at 90.3 kDa; no empirical band size or isoform masses are supplied, so verify any band near that size with identity controls.
| UniProt predicted mass | sets a 90.3 kDa reference, without establishing observed migration |
| Isoform 1 | may differ in size from other named isoforms; its mass is not supplied |
| Isoforms 2 and 3 | may migrate differently; their masses are not supplied |
| Isoforms 4 and 5 | may migrate differently; their masses are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Multiple bands | five annotated isoforms could contribute, but their migration is unknown | check band identity with CAGE1 knockdown or isoform-specific controls |
| Band higher than expected | an isoform is possible; the cause of higher migration is unestablished | verify the band with CAGE1 knockdown and a second antibody |
| Band lower than expected | an isoform is possible; the cause of lower migration is unestablished | verify the band with CAGE1 knockdown and a second antibody |
| Weak or no signal | CAGE1 abundance in the tested sample is not established | check sample loading and antibody performance with a validated positive control |
| Fragments below expected size | sample degradation or nonspecific binding is possible | prepare fresh lysate and check whether the bands disappear with CAGE1 knockdown |
Experiment evidence
Western blot images showing CAGE1 band
A12728 · CAGE1
A12728-1 · CAGE1
A12728-1 · CAGE1
A12728-1 · CAGE1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CAGE1 Western blot
🧪For positive controls for CAGE1 in Western blot, you can use no HPA-verified sample because no positive tissue or cell data were supplied.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: With no HPA data, tissue controls cannot be verified; use a knockdown or KO line for a negative control.
HPA tissue expression evidence for CAGE1
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced CAGE1 Western Blot Tips
Deeper troubleshooting and optimisation questions for CAGE1, answered from its protein features.
Where should the main CAGE1 band appear?
Band shift · The canonical sequence is 777 amino acids with a predicted mass of 90.3 kDa. No empirical band position is supplied, so use 90.3 kDa as a reference, not a guaranteed apparent mass.
Could CAGE1 isoforms produce bands of different sizes?
Isoforms · UniProt lists five isoforms with sequence differences that could change their calculated masses. Isoform 2 lacks residues 1–136; isoforms 3, 4 and 5 have changes near the C terminus. These features do not establish which isoforms are present in your sample or whether their bands resolve.
An epitope within canonical residues 137–667 avoids the listed variable regions at 1–136 and 668–777. Check the antibody's stated epitope against each isoform sequence before assuming equal recognition.
An epitope within canonical residues 137–667 avoids the listed variable regions at 1–136 and 668–777. Check the antibody's stated epitope against each isoform sequence before assuming equal recognition.
Could annotated modifications explain a shifted CAGE1 band?
PTM · The supplied features list no modified residues or glycosylation sites. They therefore provide no specific modification to assign to a shift. A difference between apparent and predicted mass needs experimental evidence to explain it.
Does this guide establish induction of CAGE1?
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 CAGE1?
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 A12728 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 CAGE1 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
How should I compare CAGE1 signal across samples?
Interpretation · First establish whether the antibody recognizes a shared sequence or an isoform-specific region. An antibody targeting canonical residues 1–136 may miss isoform 2, while one targeting changed C-terminal sequence may respond differently to isoforms 3–5. Compare signals only with the antibody's isoform coverage in mind.
Compare their apparent sizes with the five listed isoform sequences, particularly isoform 2's 1–136 deletion and the C-terminal changes in isoforms 3–5. The features alone cannot identify an observed band; confirm its identity with an appropriate isoform-aware antibody or other experimental evidence.
Compare their apparent sizes with the five listed isoform sequences, particularly isoform 2's 1–136 deletion and the C-terminal changes in isoforms 3–5. The features alone cannot identify an observed band; confirm its identity with an appropriate isoform-aware antibody or other experimental evidence.
Boster reagents
CAGE1 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-CAGE1 Antibody
Cat # A12728
Real WB data
Anti-CAGE-1 Antibody
Cat # A12728-1
Two the supplier anti-CAGE1 antibodies list Human, Mouse, and Rat reactivity and have WB images. A12728 shows HeLa, HEK293T, mouse and rat testis, and H9C2 lysates at 1:500; A12728-1 reports various cells at 1:2000 without identifying them.
Which to pick: Choose A12728 when its documented sample types match your experiment: its WB caption identifies five lysates and loading conditions. A12728-1 also has a WB image, but its caption does not identify the cells. Both list Human, Mouse, and Rat reactivity.
Source: BosterBio CAGE1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8TC20.
- Human Protein Atlas. CAGE1 tissue expression.
- PMC3994729 — target-verified WB comparison