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SPARCL1 / SPARC-like protein 1 · Western blot design guide
Design a Western Blot for SPARCL1
Source-linked SPARCL1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SPARCL1 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
SPARCL1 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 | ~75.2 kDa | |
| Observed band | Approximately 62 kDa | |
| Gel | 5–20% (catalog A05726) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked SPARCL1 Western Blot Protocol Options
The A05726 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human U-87MG, human SHG-44, human MDA-MB-231, human K562, rat C6, mouse smooth muscle (catalog A05726) |
| Gel % | 5–20% (catalog A05726) |
| Load | 50ug; reducing conditions (catalog A05726) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A05726) |
| Membrane | Nitrocellulose membrane (catalog A05726) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A05726) |
| Primary antibody | A05726; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | overnight (catalog A05726) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:10000 (catalog A05726) |
| Secondary incubation | 1.5 hour at RT (catalog A05726) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05726) |
| Detection | ECL (catalog A05726) |
Section 2
What Is the Expected SPARCL1 Western Blot Band Size?
SPARCL1 has a predicted full-length mass of 75.2 kDa and an empirical band near 62 kDa; the cause of their difference is not established.
| Band near 62 kDa | Empirical SPARCL1-reactive band; the reason it runs below the predicted mass is unknown. |
| Band near 75.2 kDa | Consistent with the predicted full-length precursor mass, pending identity controls. |
| Little or no band in whole-cell lysate | SPARCL1 is secreted into the extracellular space and matrix. |
| Band below the precursor position | Signal-peptide cleavage could reduce size, but its effect on migration is unestablished. |
| Several bands at different positions | Isoforms 1 and 2 could differ in size; distinct bands are unconfirmed. |
| Broad band or smear | Variable N- and O-linked glycosylation is possible; a visible smear is unconfirmed. |
💡Expected SPARCL1 appearanceThe predicted full-length mass is 75.2 kDa, while antibody QC reports a band near 62 kDa; the cause of the difference is unestablished, so confirm identity with an appropriate control.
| Predicted full-length mass | Provides a 75.2 kDa reference; the empirical band is near 62 kDa. |
| Signal peptide at residues 1–16 | Cleavage could make mature protein smaller than the precursor, but its apparent migration is unknown. |
| N-linked glycosylation at Asn169 | Can affect apparent mass; its contribution to migration is unknown. |
| O-linked GalNAc glycosylation at Thr31 | Can affect apparent mass; its contribution to migration is unknown. |
| Isoforms 1 and 2 | Could differ in size; their relative sizes and migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Secreted SPARCL1 may be depleted from whole-cell lysate. | Check conditioned medium or extracellular matrix alongside a positive control. |
| Band higher than expected | N- or O-linked glycosylation could affect migration. | Compare matched glycosidase-treated samples and confirm band identity. |
| Band lower than expected | The reported 62 kDa band is below the 75.2 kDa prediction; its cause is unestablished. | Confirm identity by knockdown or a second antibody recognizing another epitope. |
| Broad smear instead of sharp band | Heterogeneous N- or O-linked glycosylation is possible. | Compare matched samples before and after appropriate glycosidase treatment. |
| Multiple bands | Isoforms 1 and 2 may differ in size, although distinct migration is unconfirmed. | Use isoform-specific validation or peptide mapping to identify the bands. |
| Weak or no signal | Secretion may leave little SPARCL1 in the sampled lysate. | Test conditioned medium with a known positive sample. |
Experiment evidence
Western blot images showing SPARCL1 band
A05726 · SPARCL1
PA2088 · SPARCL1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SPARCL1 Western blot
🧪For positive controls for SPARCL1 in Western blot, you can use an independently validated SPARCL1-positive sample, since the supplied HPA evidence lists no positive tissue or cell line.
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 such as Ponceau alongside the samples.
⚠️Feasibility: Because SPARCL1 is secreted, whole-cell lysate may give weak signal; consider conditioned medium.
HPA tissue expression evidence for SPARCL1
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 SPARCL1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SPARCL1, answered from its protein features.
How should SPARCL1 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 SPARCL1 isoforms affect band interpretation?
Isoforms · UniProt lists isoforms 1 and 2; isoform 2 lacks canonical residues 1–125. An antibody targeting that region may miss isoform 2. The deletion makes isoform 2 a candidate when investigating a smaller band, but the supplied features do not identify the approximately 62 kDa band as that isoform.
Which SPARCL1 glycosylation sites matter when assessing band mobility?
PTM · In UniProt canonical numbering, O-linked sites are 31, 40, 44, 116, 331, and 398; N-linked sites are 169, 176, 196, 280, 412, and 476. Compare bands across samples with these sites in mind, but site annotations alone cannot establish glycan occupancy or a visible shift.
UniProt canonical coordinates list phosphoserines at 76, 84, 92, 171, 272, 358, 365, and 420. Use those coordinates when matching an antibody or peptide result; numbering may differ in an isoform or publication. The annotations do not show that phosphorylation causes a detectable band shift.
UniProt canonical coordinates list phosphoserines at 76, 84, 92, 171, 272, 358, 365, and 420. Use those coordinates when matching an antibody or peptide result; numbering may differ in an isoform or publication. The annotations do not show that phosphorylation causes a detectable band shift.
Does this guide establish induction of SPARCL1?
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 SPARCL1?
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 A05726 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 SPARCL1 Western blot signals be quantified?
Quantitation · Quantify the same defined band or bands across samples and report whether the material came from cells or an extracellular compartment. Because SPARCL1 has two isoforms and multiple annotated modifications, establish which signal is being measured before treating band intensities as comparable SPARCL1 abundance.
Why might SPARCL1 appear near 62 kDa instead of 75.2 kDa?
Interpretation · The 75.2 kDa prediction is for the canonical sequence, while the reported band is approximately 62 kDa. SPARCL1 has a signal peptide at residues 1–16, two isoforms, and listed glycosylation and phosphorylation sites. These features alone do not establish which form produces the 62 kDa band or explain the full difference.
SPARCL1 is annotated as secreted and located in extracellular space and extracellular matrix. Consider both extracellular material and cells when choosing samples, and record the sampled compartment so band intensities can be interpreted consistently.
Check whether the antibody epitope lies within canonical residues 1–125, which are missing from isoform 2. Also account for the 1–16 signal peptide when considering mature protein. Neither feature alone identifies an unexpected band; the listed modification sites likewise do not prove a mobility shift.
SPARCL1 is annotated as secreted and located in extracellular space and extracellular matrix. Consider both extracellular material and cells when choosing samples, and record the sampled compartment so band intensities can be interpreted consistently.
Check whether the antibody epitope lies within canonical residues 1–125, which are missing from isoform 2. Also account for the 1–16 signal peptide when considering mature protein. Neither feature alone identifies an unexpected band; the listed modification sites likewise do not prove a mobility shift.
Boster reagents
SPARCL1 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-SPARCL1 Antibody Picoband®
Cat # A05726
Real WB data
Anti-SPARC-like protein 1 SPARCL1 Antibody Picoband®
Cat # PA2088
Two the supplier anti-SPARCL1 antibodies, A05726 and PA2088, list human, mouse and rat reactivity and have Western blot images. A05726 reports a band near 62 kDa versus an expected 75 kDa; the supplied captions do not establish band identity independently.
Which to pick: Choose A05726 for a closer match to the reported U-87MG, SHG-44, MDA-MB-231, K562, rat C6 or mouse smooth muscle samples. PA2088 shows rat and mouse lung tissue and several named cell lysates. Both have WB images; match your sample to the reported lanes.
Source: BosterBio SPARCL1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14515.
- Human Protein Atlas. SPARCL1 tissue expression.
- PMC7241726 — target-verified WB comparison
- PMC13250643 — target-verified WB comparison
- PMC6030747 — target-verified WB comparison