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- Table of Contents
CERS6 / Ceramide synthase 6 · Western blot design guide
Design a Western Blot for CERS6
Source-linked CERS6 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CERS6 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
CERS6 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 | ~44.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cerebellum (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 | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked CERS6 Western Blot Protocol Options
The A07033 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat brain tissue lysate (catalog A07033) |
| Gel % | 12–15% (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 | A07033 · (A) 1 and (B) 2 μg/mL (catalog A07033) |
| 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 CERS6 Western Blot Band Size?
CERS6 has a predicted monomer mass of 44.9 kDa; glycosylation, isoforms, and homodimer formation could affect bands, but their migration effects are unproven.
| Band near 44.9 kDa | Consistent with the predicted CERS6 monomer; confirm identity with controls. |
| Band near 90 kDa | Could represent a homodimer if it survives electrophoresis; this migration is unverified. |
| Slight upward shift from 44.9 kDa | Could reflect glycosylation at Asn18 if occupied; a visible shift is unproven. |
| Multiple bands at different positions | Could reflect isoforms 1 and 2; distinct migration is unproven. |
| Weak band in soluble lysate fraction | CERS6 is an endoplasmic reticulum membrane protein and may remain in the membrane fraction. |
💡Expected CERS6 appearanceUniProt predicts a 44.9 kDa CERS6 monomer; glycosylation, isoforms, and homodimer formation may affect migration, but no empirical band size is supplied, so confirm band identity with controls.
| Predicted monomer mass | 44.9 kDa is the sequence-based reference size, not a measured band. |
| N-linked glycosylation site at Asn18 | Occupancy could raise apparent size, but a visible shift is not established. |
| Homodimer formation | Could yield a band near twice the monomer size if the dimer survives electrophoresis. |
| Isoforms 1 and 2 | Could differ in size, but their masses and separation on a blot are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CERS6 is a multi-pass endoplasmic reticulum membrane protein that may be poorly recovered. | Check membrane extraction and probe a membrane-enriched fraction. |
| Band higher than expected | A homodimer may persist during electrophoresis, or Asn18 may be glycosylated. | Compare denaturing conditions and use a deglycosylation control to investigate identity. |
| Band lower than expected | Isoform 2 may differ in size from isoform 1; their relative masses are unknown. | Check isoform expression and confirm the band with an independent antibody or CERS6 depletion. |
| Multiple bands | Isoforms 1 and 2 or a persistent homodimer are possible; distinct bands are unverified. | Compare denaturing conditions and use CERS6 depletion to identify specific bands. |
| Weak or no signal | Recovery or solubilization of this multi-pass membrane protein may be insufficient. | Assess membrane-fraction recovery and include a positive lysate control. |
Experiment evidence
Western blot images showing CERS6 band
A07033 · CERS6
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CERS6 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CERS6 in Western blot, you can use cerebellum, where HPA reports high expression.
Positive control: Cerebellum (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: As an ER membrane protein, CERS6 may require effective membrane solubilization for detection.
HPA tissue expression evidence for CERS6
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 |
|---|---|---|---|---|
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CERS6 Western Blot Tips
Deeper troubleshooting and optimisation questions for CERS6, answered from its protein features.
How should CERS6 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 CERS6 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. At canonical UniProt position 334, isoform 2 replaces K with KAGKWNPLH. This sequence difference could affect mass, but the supplied features do not establish whether the isoforms resolve as separate bands.
Could glycosylation affect the CERS6 band?
PTM · UniProt annotates one N-linked glycosylation site at asparagine 18, using UniProt coordinates. Compare matched samples before and after an appropriate N-glycan removal treatment if assessing its contribution. The annotation alone does not show that glycosylation causes a visible band shift.
Does this guide establish induction of CERS6?
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 CERS6 Western blot?
Transfer · CERS6 is a 384-residue, multi-pass ER membrane protein with a predicted mass of 44.9 kDa. Check transfer efficiency for the CERS6 size region and optimize membrane-protein handling if recovery is weak. The supplied features do not specify a preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A07033 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 CERS6 bands be quantified?
Quantitation · Quantify the same defined band region across samples and document whether it represents a presumed monomer or a higher band. UniProt reports both monomer and homodimer, while supplying no observed Western blot positions; avoid combining distinct bands without establishing their identity.
Should CERS6 run at its predicted 44.9 kDa?
Interpretation · 44.9 kDa is the predicted mass; no observed Western blot band position is supplied. CERS6 is a multi-pass ER membrane protein with an annotated N-linked glycosylation site, but these features alone do not establish a visible shift or explain any difference from 44.9 kDa.
Consider the annotated isoform-2 sequence change at canonical UniProt position 334, N-linked glycosylation at asparagine 18, and reported monomer and homodimer states. None establishes the identity of an unexpected Western blot band. Compare its apparent size and behavior across matched conditions before assigning it to CERS6.
Consider the annotated isoform-2 sequence change at canonical UniProt position 334, N-linked glycosylation at asparagine 18, and reported monomer and homodimer states. None establishes the identity of an unexpected Western blot band. Compare its apparent size and behavior across matched conditions before assigning it to CERS6.
Boster reagents
CERS6 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-LASS6 CERS6 Antibody
Cat # A07033
The catalog reports one anti-CERS6 antibody, A07033 (Anti-LASS6 CERS6), with reported Human, Mouse, and Rat reactivity. Its Western blot image uses rat brain tissue lysate at 1 and 2 μg/mL; the supplied evidence shows no Human or Mouse WB examples.
Which to pick: A07033 is the only listed option. Its supplied WB image documents rat brain lysate at 1 and 2 μg/mL. Human and Mouse are listed as reactive, but no WB examples for those samples are provided.
Source: BosterBio CERS6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q6ZMG9.
- Human Protein Atlas. CERS6 tissue expression.
- PMC13369121 — target-verified WB comparison
- PMC8253294 — target-verified WB comparison