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- Table of Contents
CST7 / Cystatin-F · Western blot design guide
Design a Western Blot for CST7
Source-linked CST7 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CST7 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
CST7 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 | ~16.5 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Bone marrow (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Reducing-condition controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked CST7 Western Blot Protocol Options
The A08883 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A08883) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A08883) |
| Primary antibody | A08883 · 1:1000 (catalog A08883) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A08883) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A08883) |
Section 2
What Is the Expected CST7 Western Blot Band Size?
Cystatin-F has a predicted precursor mass of 16.5 kDa; cleavage, glycosylation, and dimerization could affect migration, but no empirical band size is supplied.
| Band near 16.5 kDa | Compatible with the predicted precursor mass; identity requires validation. |
| Band above 16.5 kDa | N-linked glycosylation may affect migration, but no shift is established. |
| Band near twice the monomer size under nonreducing conditions | Compatible with the disulfide-linked homodimer. |
| Band below 16.5 kDa | Signal-peptide removal may yield a smaller mature protein. |
| Little or no band in whole-cell lysate | Secretion may reduce the cellular signal. |
💡Expected CST7 appearanceThe predicted precursor mass is 16.5 kDa; signal-peptide cleavage, N-linked glycosylation, and disulfide-linked dimerization may alter migration, but no empirical band size is supplied, so validate band identity with appropriate controls.
| Predicted precursor mass | 16.5 kDa provides a sequence-based reference, not an observed band. |
| N-linked glycosylation at Asn62 | May increase apparent size; the shift is not established. |
| N-linked glycosylation at Asn115 | May increase apparent size; the shift is not established. |
| Disulfide-linked homodimer | May produce a band near twice the monomer size under nonreducing conditions. |
| Signal-peptide cleavage at residues 1–19 | May make the mature protein smaller than the precursor. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cystatin-F is secreted as well as cytoplasmic. | Check conditioned medium alongside lysate and include a positive control. |
| Band higher than expected | A disulfide-linked homodimer or N-linked glycosylation may alter migration. | Compare reducing and nonreducing samples; use a glycosidase control to assess glycosylation. |
| Band lower than expected | Removal of the signal peptide may reduce mass. | Compare with the precursor reference and confirm band identity with a specificity control. |
| Broad smear instead of sharp band | Variable N-linked glycosylation at the two annotated sites is possible. | Compare untreated and glycosidase-treated samples to test that explanation. |
| Multiple bands | Mature and precursor forms or monomer and disulfide-linked dimer may coexist. | Compare reducing and nonreducing lanes and verify each band with a specificity control. |
| Weak or no signal | Secretion may limit the amount retained in cells. | Test conditioned medium and a known positive sample. |
Experiment evidence
Western blot images showing CST7 band
A08883 · CST7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CST7 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CST7 in Western blot, you can use bone marrow, the top HPA positive tissue.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Because CST7 is secreted as well as cytoplasmic, consider conditioned medium if whole-cell lysate signal is weak.
HPA tissue expression evidence for CST7
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 |
|---|---|---|---|---|
| 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CST7 Western Blot Tips
Deeper troubleshooting and optimisation questions for CST7, answered from its protein features.
How should CST7 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.
Are CST7 isoforms expected to explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. Do not assign multiple bands to annotated isoforms on this evidence alone; consider signal-peptide processing, glycosylation, and disulfide-linked dimerization when designing follow-up comparisons.
Can glycosylation help interpret CST7 bands?
PTM · CST7 has N-linked glycosylation sites at UniProt positions 62 and 115. Compare treated and untreated samples if testing whether glycans contribute to a band pattern. The listed sites alone do not show that a visible shift will occur.
Does this guide establish induction of CST7?
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 CST7?
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 A08883 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 CST7 Western blot signals be quantified?
Quantitation · Compare signals from the same sample fraction and preparation conditions. CST7 is listed as secreted and cytoplasmic, with N-linked glycosylation sites and a disulfide-linked homodimer; changing the fraction or reducing conditions can change which forms are measured.
Why might CST7 migrate differently from its predicted 16.5 kDa mass?
Interpretation · The 16.5 kDa prediction is for the full-length protein. CST7 has a signal peptide at residues 1–19 and N-linked glycosylation sites at UniProt positions 62 and 115. These features could affect the protein detected, but the supplied record gives no observed band size, so it cannot establish a shift or its cause.
UniProt describes CST7 as a disulfide-linked homodimer. Compare reducing and nonreducing sample conditions when assessing a higher band. Four disulfide bonds are listed, but these features alone do not identify an observed band.
UniProt lists CST7 in secreted and cytoplasmic locations. If the experiment calls for both pools, examine conditioned medium and cell lysate separately, with consistent collection conditions. A band in either fraction should be interpreted in light of those listed locations.
Compare cell lysate with conditioned medium, and reducing with nonreducing preparations. CST7 has a signal peptide at residues 1–19, N-linked glycosylation sites at UniProt positions 62 and 115, and a disulfide-linked homodimer. These are candidates to test, not established explanations for any particular band.
UniProt describes CST7 as a disulfide-linked homodimer. Compare reducing and nonreducing sample conditions when assessing a higher band. Four disulfide bonds are listed, but these features alone do not identify an observed band.
UniProt lists CST7 in secreted and cytoplasmic locations. If the experiment calls for both pools, examine conditioned medium and cell lysate separately, with consistent collection conditions. A band in either fraction should be interpreted in light of those listed locations.
Compare cell lysate with conditioned medium, and reducing with nonreducing preparations. CST7 has a signal peptide at residues 1–19, N-linked glycosylation sites at UniProt positions 62 and 115, and a disulfide-linked homodimer. These are candidates to test, not established explanations for any particular band.
Boster reagents
CST7 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-Cystatin-F CST7 Antibody
Cat # A08883
the supplier A08883 is an anti-CST7 antibody listed for human and mouse reactivity. Its Western blot image uses cell-line extracts at 25 µg per lane and a 1:1000 primary dilution; the supplied caption does not identify the cell lines.
Which to pick: A08883 is the only listed CST7 antibody and has a Western blot image. Check that its stated human or mouse reactivity matches your sample; the image caption does not specify which cell lines were tested.
Source: BosterBio CST7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O76096.
- Human Protein Atlas. CST7 tissue expression.
- PMC12140383 — target-verified WB comparison
- PMC10037308 — target-verified WB comparison