This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SLIT3 / Slit homolog 3 protein · Western blot design guide
Design a Western Blot for SLIT3
Source-linked SLIT3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLIT3 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
SLIT3 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 | ~167.7 kDa | |
| Observed band | ~145 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (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 + Cleaved | |
| Caveat | Observed below predicted | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked SLIT3 Western Blot Protocol Options
The A04745 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | nitrocellulose (catalog A04745) |
| Membrane | nitrocellulose (catalog A04745) |
| Blocking | After blocking the membrane was probed with the primary antibody diluted to 1:800 (catalog A04745) |
| Primary antibody | A04745 · 1:800 (catalog A04745) |
| 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 SLIT3 Western Blot Band Size?
SLIT3 has a predicted precursor size of 167.7 kDa and a reported ~145 kDa band; the cause of their difference is not established.
| Predominant band at ~145 kDa | Empirical SLIT3 band reported in bovine thyroid lysate; confirm identity with controls. |
| Band near 167.7 kDa | Could represent SLIT3 near its predicted precursor mass; confirm identity. |
| Band below precursor size | Could reflect signal-peptide cleavage; the resulting migration is not established. |
| Little or no band in whole-cell lysate | SLIT3 is secreted and may be depleted from cells. |
| Multiple bands | Isoforms 1, 2, 3, and 4 exist, but distinct band positions are not established. |
| Broad band or smear | Could reflect variation in N-linked glycosylation; visible heterogeneity is not established. |
💡Expected SLIT3 appearanceSLIT3 has a predicted 167.7 kDa precursor and a reported predominant ~145 kDa band in bovine thyroid lysate; the size difference is unexplained, so confirm band identity with appropriate controls.
| Predicted precursor mass | 167.7 kDa by sequence; the reported ~145 kDa band has no established explanation for its lower migration. |
| Signal peptide at residues 1–33 | Cleavage can make mature SLIT3 smaller than its precursor, but its apparent band size is unknown. |
| N-linked glycosylation at Asn72, Asn192, Asn563, Asn622, Asn784, Asn792, Asn797, and Asn928 | Glycan occupancy could alter apparent migration; the size and visibility of any effect are unknown. |
| Isoforms 1, 2, 3, and 4 | Alternative sequences may differ in size, but their masses and band positions are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Secreted SLIT3 may be scarce in the cell fraction. | Check conditioned medium and a positive-control lysate. |
| Band higher than expected | N-linked glycosylation or retained signal peptide could affect migration. | Compare with a positive control and assess glycosylation or processing experimentally. |
| Band lower than expected | The reported ~145 kDa band is below the 167.7 kDa prediction, but the cause is unknown. | Compare with the reported positive lysate and confirm identity using an independent antibody or knockdown. |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but unproven. | Compare treated and untreated samples after enzymatic deglycosylation. |
| Multiple bands | Alternative isoforms are documented, but their migration is unknown. | Check band identity with knockdown or isoform-specific reagents if available. |
| Weak or no signal | Secretion may leave little SLIT3 in the sampled fraction. | Examine conditioned medium and include a positive-control sample. |
Experiment evidence
Western blot images showing SLIT3 band
A04745 · SLIT3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLIT3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SLIT3 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (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: Because SLIT3 is secreted, whole-cell lysates may show little signal; consider an extracellular fraction.
HPA tissue expression evidence for SLIT3
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 | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | ciliated cells (ciliary rootlets) | Medium | Protein (IHC) | HPA → |
| Colon | glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | glandular 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lung | alveolar cells | Not detected | Protein (IHC) | HPA → |
| Nasopharynx | basal cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLIT3 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLIT3, answered from its protein features.
How should SLIT3 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 can SLIT3 isoforms affect band interpretation?
Isoforms · UniProt lists four isoforms. Relative to the canonical sequence, isoform 2 lacks residues 1117–1216, isoform 3 replaces residues 1446–1523, and isoform 4 changes residue 906 from K to KVLWFCCP. Check whether the antibody epitope is retained in each isoform before assigning bands. These are UniProt canonical-sequence coordinates; antibody numbering may differ.
Which SLIT3 glycosylation sites matter when assessing band variation?
PTM · UniProt annotates N-linked sites at residues 72, 192, 563, 622, 784, 792, 797, 928, 1008, 1025, 1181, 1247, and 1406. A comparison before and after N-glycan removal could test whether glycosylation contributes to band variation. The listed sites alone do not establish a visible shift. Coordinates follow the UniProt canonical sequence and may differ from antibody numbering.
Does this guide establish induction of SLIT3?
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 SLIT3 Western blot?
Transfer · SLIT3 is a large protein: its predicted mass is 167.7 kDa, and the reported apparent band is about 145 kDa. Start with conditions suited to transferring proteins in that size range, then check the membrane and post-transfer gel for retained protein. Optimize transfer time if recovery is poor.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04745 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 can SLIT3 bands be quantified consistently?
Quantitation · Quantify the same antibody-reactive band and sample fraction across lanes, documenting whether it is the reported approximately 145 kDa band. SLIT3 is secreted and has four annotated isoforms, so mixing medium with lysate or combining distinct bands could obscure comparisons. Verify that the measured signal is within the assay’s linear range.
Why might SLIT3 migrate near 145 kDa instead of 167.7 kDa?
Interpretation · The reported apparent band is about 145 kDa, while the predicted mass is 167.7 kDa. SLIT3 has a signal peptide at residues 1–33, 13 annotated N-linked glycosylation sites, and four isoforms. These features warrant checking which form the antibody detects, but their presence alone does not explain the mass difference.
UniProt annotates SLIT3 as secreted and identifies a signal peptide at residues 1–33. Check conditioned medium as well as cell lysate when evaluating expression. Keep the fractions separate when comparing band intensity.
First compare their sizes and antibody-epitope coverage with the four annotated isoforms. Then consider the signal peptide at residues 1–33 and the 13 N-linked glycosylation sites. SLIT3 also has 37 annotated disulfide bonds, so compare reducing conditions consistently. None of these features alone identifies an unexpected band.
UniProt annotates SLIT3 as secreted and identifies a signal peptide at residues 1–33. Check conditioned medium as well as cell lysate when evaluating expression. Keep the fractions separate when comparing band intensity.
First compare their sizes and antibody-epitope coverage with the four annotated isoforms. Then consider the signal peptide at residues 1–33 and the 13 N-linked glycosylation sites. SLIT3 also has 37 annotated disulfide bonds, so compare reducing conditions consistently. None of these features alone identifies an unexpected band.
Boster reagents
SLIT3 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-SLIT-3 Antibody
Cat # A04745
the supplier A04745 is an anti-SLIT3 antibody with a Western blot image showing a predominant ~145 kDa band in bovine thyroid whole-cell lysate. The supplied evidence documents this tested sample and does not establish Western blot performance in other tissues or species.
Which to pick: A04745 is the only listed option. Its stated reactivity includes bovine, canine, human, mouse, and rat, but the supplied Western blot image uses bovine thyroid lysate. Use that tested context to guide selection and verify performance in your own sample.
Source: BosterBio SLIT3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O75094.
- Human Protein Atlas. SLIT3 tissue expression.
- PMC13176150 — target-verified WB comparison
- PMC13071080 — target-verified WB comparison
- PMC8974015 — target-verified WB comparison