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- Table of Contents
SLIT2 / Slit homolog 2 protein · Western blot design guide
Design a Western Blot for SLIT2
Real validated SLIT2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLIT2 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
SLIT2 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~169.9 kDa | |
| Observed band | ≈170 kDa | |
| Gel | 5–20% (catalog A01627-1) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Cleavage-fragment controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated SLIT2 Western Blot Protocols
The A01627-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human Raji, human PC-3 (catalog A01627-1) |
| Gel % | 5–20% (catalog A01627-1) |
| Load | 30 ug; reducing conditions (catalog A01627-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01627-1) |
| Membrane | nitrocellulose membrane (catalog A01627-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01627-1) |
| Primary antibody | A01627-1 · 0.5 μg/mL (catalog A01627-1) |
| Primary incubation | overnight at 4°C (catalog A01627-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01627-1) |
| Secondary incubation | 1.5 hour at RT (catalog A01627-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01627-1) |
| Detection | ECL (catalog A01627-1) |
Section 2
What Is the Expected SLIT2 Western Blot Band Size?
SLIT2 is predicted at 169.9 kDa and observed near 170 kDa; the small difference has no established cause in the supplied evidence.
| Band near 170 kDa | matches the empirical SLIT2 band and its 169.9 kDa predicted mass; confirm identity with antibody controls |
| Higher band near twice the monomer size | could reflect SLIT2 homodimer retained under nonreducing conditions |
| Band below 170 kDa | could reflect signal peptide removal or the annotated C-terminal cleavage product; fragment size is unspecified |
| Broad band or smear near the main band | could reflect heterogeneous N-linked glycosylation |
| Several bands at different positions | could reflect cleavage products or isoforms 1, 2, and 3; distinct isoform migration is unproven |
| Little or no band in whole-cell lysate | SLIT2 is secreted and may be depleted from the lysate |
💡Expected SLIT2 appearanceSLIT2 has a predicted mass of 169.9 kDa and an empirical band near 170 kDa; confirm band identity with appropriate antibody controls because its annotated modifications and processing do not establish a specific migration change.
| Predicted SLIT2 mass | 169.9 kDa for the full-length sequence, consistent with the empirical band near 170 kDa |
| N-linked glycosylation at Asn66, Asn186, Asn564, Asn623, Asn794, Asn799, Asn1009, and Asn1010 | may alter apparent migration if occupied; no shift magnitude is established |
| Signal peptide at residues 1–30 | cleavage yields a smaller mature chain than the full-length precursor; apparent size is unspecified |
| SLIT2 homodimer | may yield a band near twice the monomer size if the dimer remains intact during electrophoresis |
| Isoforms 1, 2, and 3 | may differ in size, but their masses and resolvable migration differences are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SLIT2 is secreted | check conditioned medium alongside lysate |
| Band higher than expected | SLIT2 homodimer may persist during electrophoresis | compare reducing and nonreducing preparations |
| Band lower than expected | signal peptide removal or annotated C-terminal cleavage | check antibody epitope and compare lysate with conditioned medium |
| Broad smear instead of sharp band | heterogeneous N-linked glycosylation is possible | compare untreated and deglycosylated samples |
| Multiple bands | cleavage products or isoforms may be present | compare antibodies targeting different SLIT2 regions |
| Weak or no signal | secreted SLIT2 may be scarce in the sampled fraction | test conditioned medium and a known positive control |
| Fragments below expected size | SLIT2 has an annotated C-terminal cleavage product | check antibody epitope and compare fragment patterns across sample fractions |
Experiment evidence
Western blot images showing SLIT2 band
A01627-1 · SLIT2
M01627 · SLIT2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLIT2 Western blot
🧪For positive controls for SLIT2 in Western blot, you can use conditioned medium from a verified SLIT2-expressing sample; the supplied HPA data identify 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: SLIT2 is secreted, so conditioned medium may give a clearer signal than whole-cell lysate.
HPA tissue expression evidence for SLIT2
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 SLIT2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLIT2, answered from its protein features.
How should SLIT2 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.
Could SLIT2 isoforms produce different bands?
Isoforms · Three isoforms are listed. Relative to the canonical sequence, isoform 2 replaces S258 with SDEEE and lacks residues 480–487; isoform 3 also lacks residues 480–487. Compare the antibody’s binding region with these changes before assigning a band to an isoform. The features do not specify each isoform’s apparent mass.
Which SLIT2 glycosylation sites matter when interpreting bands?
PTM · UniProt lists N-linked sites at positions 66, 186, 564, 623, 794, 799, 1009, 1010, 1019, 1183, 1266, and 1300. These are UniProt canonical-sequence coordinates; antibody or paper numbering may differ. Glycosylation could affect migration, but the site list alone does not establish a particular band shift.
Does this guide establish induction of SLIT2?
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 SLIT2 Western blot?
Transfer · SLIT2 is predicted at 169.9 kDa, so choose transfer conditions validated for proteins around 170 kDa. Check transfer completeness, including whether protein remains in the gel, before interpreting a weak or absent band. The supplied features do not establish one required transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01627-1 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 I quantify SLIT2 across sample fractions?
Quantitation · Quantify comparable sample fractions consistently. SLIT2 is secreted, and its reported cleavage products differ in diffusibility and cell association, so a change in one fraction or band need not represent a change in total SLIT2. Define which band and fraction you measure before comparing samples.
Does the ≈170 kDa band indicate a SLIT2 mass shift?
Interpretation · The supplied apparent band (≈170 kDa) is close to the predicted 169.9 kDa. SLIT2 has a signal peptide at residues 1–30 and 12 annotated N-linked glycosylation sites, but these features alone do not establish a visible shift or explain any mass difference.
Yes. SLIT2 is secreted, and the supplied location note says its C-terminal cleavage protein is more diffusible while the larger N-terminal protein is more tightly cell associated. Examine the relevant sample fractions when assessing signal or missing bands; the features do not identify the cleavage enzyme or fragment sizes.
Consider the three annotated isoforms and the reported cleavage into a larger N-terminal and more diffusible C-terminal protein. Check which SLIT2 region the antibody recognizes and which sample fraction contains the band. These features do not establish the identity or size of any particular lower band.
Yes. SLIT2 is secreted, and the supplied location note says its C-terminal cleavage protein is more diffusible while the larger N-terminal protein is more tightly cell associated. Examine the relevant sample fractions when assessing signal or missing bands; the features do not identify the cleavage enzyme or fragment sizes.
Consider the three annotated isoforms and the reported cleavage into a larger N-terminal and more diffusible C-terminal protein. Check which SLIT2 region the antibody recognizes and which sample fraction contains the band. These features do not establish the identity or size of any particular lower band.
Boster reagents
SLIT2 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-SLIT2 Antibody Picoband®
Cat # A01627-1
Real WB data
Anti-Slit2 Monoclonal Antibody
Cat # M01627
Two the supplier anti-SLIT2 antibodies have WB images. A01627-1 shows an approximately 170 kDa band in four human cell lysates under reported conditions. M01627 shows a blot of SH-SY5Y lysate, but its caption gives no band size or assay conditions.
Which to pick: Choose A01627-1 for human samples when a documented 170 kDa WB example is useful. M01627 lists human, mouse, and rat reactivity, but its supplied WB example uses only SH-SY5Y lysate; the image does not establish WB performance in mouse or rat.
Source: BosterBio SLIT2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O94813.
- Human Protein Atlas. SLIT2 tissue expression.
- PMC6745244 — target-verified WB comparison
- PMC9999516 — target-verified WB comparison
- PMC13102090 — target-verified WB comparison