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KSR1 / Kinase suppressor of Ras 1 · Western blot design guide
Design a Western Blot for KSR1
Source-linked KSR1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KSR1 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
KSR1 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 | ~102.2 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Ovary (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 C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked KSR1 Western Blot Protocol Options
The M03285 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HEK293 cell lysate (catalog M03285) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M03285; 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 KSR1 Western Blot Band Size?
KSR1 is predicted at 102.2 kDa; isoforms, phosphorylation, and homodimerization could affect its pattern, but their electrophoretic effects are unestablished here.
| Band near 102.2 kDa | consistent with the UniProt predicted mass; confirm KSR1 identity with controls |
| Band near twice the predicted mass | could reflect a retained KSR1 homodimer; SDS conditions may disrupt it |
| Several bands near the expected region | could include KSR1 isoforms 1, 2, 3, and 4; their separation is unestablished |
| Close doublet near the expected region | could reflect phosphorylation states; confirm with phosphatase treatment |
💡Expected KSR1 appearanceUniProt predicts KSR1 at 102.2 kDa, but no empirical band size is supplied; use a positive control and KSR1 depletion to verify any candidate band.
| UniProt predicted mass | places the reference band near 102.2 kDa |
| KSR1 homodimer | could appear near twice the monomer size if it survives sample preparation |
| Splice isoforms 1 and 2 | may differ in size; their individual masses and migration are unspecified |
| Splice isoforms 3 and 4 | may differ in size; their individual masses and migration are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | KSR1 abundance or recovery may be low in the sampled fraction | check a positive control and assess cytoplasmic and membrane fractions |
| Band higher than expected | a KSR1 homodimer may persist during preparation | compare denaturing conditions and confirm identity by KSR1 depletion |
| Band lower than expected | a smaller splice isoform may be present | compare isoform expression and verify the band by KSR1 depletion |
| Multiple bands | isoforms or phosphorylation states may contribute | compare isoform expression, phosphatase treatment, and KSR1 depletion |
| Weak or no signal | KSR1 recovery or expression may be insufficient | check loading, antibody performance, and a KSR1 positive control |
| Fragments below expected size | sample degradation may produce smaller immunoreactive fragments | prepare fresh lysate with protease inhibitors and confirm identity by KSR1 depletion |
Experiment evidence
Western blot images showing KSR1 band
M03285 · KSR1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KSR1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for KSR1 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Ovary (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: KSR1 is cytoplasmic and membrane-associated, so tissue lysates should be suitable.
HPA tissue expression evidence for KSR1
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 | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | glandular cells | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced KSR1 Western Blot Tips
Deeper troubleshooting and optimisation questions for KSR1, answered from its protein features.
How should KSR1 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 KSR1 isoforms affect band interpretation?
Isoforms · UniProt lists four isoforms. Isoforms 3 and 4 lack canonical residues 1–137. Isoform 2 replaces residues 504–526 with D; isoforms 2 and 4 replace residues 898–923 with EL. These sequence differences could affect recognition or migration, but they do not establish where bands will appear.
An epitope within canonical residues 1–137 would be absent from isoforms 3 and 4. Residues 504–526 differ in isoform 2, and residues 898–923 differ in isoforms 2 and 4. Check the antibody epitope against the isoforms you intend to measure.
An epitope within canonical residues 1–137 would be absent from isoforms 3 and 4. Residues 504–526 differ in isoform 2, and residues 898–923 differ in isoforms 2 and 4. Check the antibody epitope against the isoforms you intend to measure.
Which KSR1 phosphorylation sites are documented?
PTM · In canonical UniProt coordinates, threonines 270, 274, and 425 and serines 311, 334, 351, 406, 569, and 888 are phosphorylated. Use these coordinates when interpreting site-specific antibodies; numbering in a paper or antibody description may follow a different sequence convention.
Does this guide establish induction of KSR1?
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 KSR1 Western blot?
Transfer · The supplied features identify a predicted 102.2 kDa canonical protein but do not specify a transfer method or settings. Check transfer efficiency around that mass and account for possible isoform bands when assessing the membrane.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03285 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 KSR1 bands be quantified?
Quantitation · Define whether the measurement targets total KSR1, a particular isoform, or a phosphorylation site. Confirm that the antibody recognizes the intended sequences, then quantify the corresponding bands consistently. KSR1 is reported in the cytoplasm and at membranes, so compare like sample preparations.
Why might KSR1 migrate away from its predicted 102.2 kDa mass?
Interpretation · 102.2 kDa is the predicted mass of the canonical sequence. KSR1 has alternative isoforms and documented phosphorylation, but these features alone cannot establish a visible shift or explain a measured difference. No empirical band position was supplied.
Compare bands with the four annotated isoforms and the antibody epitope. KSR1 also has nine documented phosphorylation sites, but their presence alone does not identify a shifted band. Because no observed band position is supplied, band identity requires experimental validation.
KSR1 is annotated in the cytoplasm and as a peripheral membrane protein, including at the cell membrane. A cytoplasmic fraction and a membrane-enriched preparation may therefore yield different KSR1 signals. Keep fraction preparation consistent across samples used for comparison.
Compare bands with the four annotated isoforms and the antibody epitope. KSR1 also has nine documented phosphorylation sites, but their presence alone does not identify a shifted band. Because no observed band position is supplied, band identity requires experimental validation.
KSR1 is annotated in the cytoplasm and as a peripheral membrane protein, including at the cell membrane. A cytoplasmic fraction and a membrane-enriched preparation may therefore yield different KSR1 signals. Keep fraction preparation consistent across samples used for comparison.
Boster reagents
KSR1 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-KSR1 Rabbit Monoclonal Antibody
Cat # M03285
M03285 is a rabbit monoclonal anti-KSR1 antibody listed for Western blotting with reported Human and Mouse reactivity. Its WB image shows KSR1 expression in HEK293 cell lysate; no publication evidence or mouse sample image is supplied.
Which to pick: M03285 is the only listed KSR1 antibody. Its HEK293 lysate WB image provides a tested context for human cell work; Mouse reactivity is listed, but no mouse WB example is supplied.
Source: BosterBio KSR1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8IVT5.
- Human Protein Atlas. KSR1 tissue expression.
- PMC3038243 — target-verified WB comparison
- PMC12133430 — target-verified WB comparison
- PMC3242779 — target-verified WB comparison