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- Table of Contents
CXCR1 / C-X-C chemokine receptor type 1 · Western blot design guide
Design a Western Blot for CXCR1
Real validated CXCR1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CXCR1 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
CXCR1 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 | ~39.8 kDa | |
| Observed band | ~50 kDa | |
| Gel | 5–20% (catalog PA2080) | |
| Positive control | Spleen (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 | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated CXCR1 Western Blot Protocols
The PA2080 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat (catalog PA2080) |
| Gel % | 5–20% (catalog PA2080) |
| Load | 30 ug; reducing conditions (catalog PA2080) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog PA2080) |
| Membrane | nitrocellulose membrane (catalog PA2080) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog PA2080) |
| Primary antibody | PA2080 · 0.5 μg/mL (catalog PA2080) |
| Primary incubation | overnight at 4°C (catalog PA2080) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog PA2080) |
| Secondary incubation | 1.5 hour at RT (catalog PA2080) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog PA2080) |
| Detection | ECL (catalog PA2080) |
Section 2
What Is the Expected CXCR1 Western Blot Band Size?
CXCR1 is predicted at 39.8 kDa and observed near 50 kDa on a reducing blot; the cause of the difference is not established.
| Band near 50 kDa | Empirical CXCR1 band in reducing whole-cell lysate; confirm identity with controls |
| Band near 40 kDa | Near the 39.8 kDa predicted mass; identity requires confirmation |
| Band above 40 kDa | N-linked glycosylation at N3 or N16 could affect migration, but its contribution is unestablished |
| Faint or absent band in soluble lysate | CXCR1 is a multipass cell-membrane protein |
💡Expected CXCR1 appearanceCXCR1 has a predicted mass of 39.8 kDa, while an antibody QC blot shows approximately 50 kDa; the cause of the difference is unestablished, so confirm band identity with appropriate controls.
| Predicted 39.8 kDa mass | Sets the sequence-based reference; the empirical band is approximately 50 kDa |
| N-linked glycosylation site N3 | Could affect apparent size if occupied; its contribution is unmeasured |
| N-linked glycosylation site N16 | Could affect apparent size if occupied; its contribution is unmeasured |
| Combined N3 and N16 glycosylation sites | Occupancy could alter migration, but no specific shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated CXCR1 may be poorly recovered | Check a membrane-enriched fraction and an extraction control |
| Band higher than expected | The observed CXCR1 band is near 50 kDa rather than the predicted 39.8 kDa; the cause is unestablished | Compare with the QC band and verify identity using a positive control or independent antibody |
| Band lower than expected | Identity or protein integrity is uncertain | Check sample handling and confirm the band with an independent antibody |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but unproven | Test N-deglycosylation alongside an untreated sample and verify band identity |
| Multiple bands | CXCR1 has no listed alternative isoforms; additional bands have uncertain identity | Compare with a positive control and confirm using an independent antibody |
| Weak or no signal | Membrane-protein recovery or CXCR1 abundance may be low | Check membrane extraction, loading, and a CXCR1-positive control |
Experiment evidence
Western blot images showing CXCR1 band
PA2080 · CXCR1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CXCR1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CXCR1 in Western blot, you can use spleen tissue.
Positive control: Spleen (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: As a multi-pass membrane protein, CXCR1 may require effective membrane protein extraction for detection.
HPA tissue expression evidence for CXCR1
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 |
|---|---|---|---|---|
| Spleen | cells in red pulp | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CXCR1 Western Blot Tips
Deeper troubleshooting and optimisation questions for CXCR1, answered from its protein features.
How should CXCR1 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 a second CXCR1 band represent a listed isoform?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It therefore provides no specific alternative isoform to assign to a second band. Investigate glycosylation and sample handling before assigning its identity.
Which CXCR1 glycosylation sites matter when interpreting bands?
PTM · UniProt lists N-linked glycosylation at Asn3 and Asn16, using UniProt sequence coordinates. Check the numbering convention used by your antibody or other reference before comparing sites. The listed sites do not establish how many bands will appear.
Does this guide establish induction of CXCR1?
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 CXCR1 Western blot?
Transfer · CXCR1 is a multi-pass cell-membrane protein. Verify recovery in the membrane-containing sample and check transfer efficiency across the observed approximately 50 kDa region. The supplied features do not establish a particular transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PA2080 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 CXCR1 bands be quantified?
Quantitation · Use the same band definition across samples and report whether quantification covers the approximately 50 kDa band alone or additional CXCR1-reactive bands. Glycosylation at Asn3 and Asn16 may affect mobility, but the supplied features do not establish which bands contain CXCR1.
Why might CXCR1 appear near 50 kDa instead of 39.8 kDa?
Interpretation · The observed band is about 50 kDa, while the predicted mass is 39.8 kDa. UniProt lists N-linked glycosylation at Asn3 and Asn16, which could affect apparent mobility. Those features alone do not establish the cause of the difference; compare untreated and deglycosylated samples to test their contribution.
UniProt reports interactions with IL8 and GNAI2, but does not establish that these complexes survive Western-blot sample preparation or produce specific bands. Do not assign a high-mass band to either interaction from its position alone.
UniProt lists one disulfide bond. Compare bands under consistent reducing and nonreducing sample conditions if a mobility difference is in question. The listed bond alone does not predict the direction or size of a shift.
UniProt reports interactions with IL8 and GNAI2, but does not establish that these complexes survive Western-blot sample preparation or produce specific bands. Do not assign a high-mass band to either interaction from its position alone.
UniProt lists one disulfide bond. Compare bands under consistent reducing and nonreducing sample conditions if a mobility difference is in question. The listed bond alone does not predict the direction or size of a shift.
Boster reagents
CXCR1 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-CXCR1 Antibody Picoband®
Cat # PA2080
PA2080 is the sole listed anti-CXCR1 antibody, with reported human reactivity and a Western blot image from human Jurkat whole-cell lysate. The caption reports a band near 50 kDa, versus an expected 40 kDa; evidence is limited to this example.
Which to pick: PA2080 is the only listed option. Its Western blot image uses human Jurkat lysate under reducing conditions, so it offers a documented starting point for human samples. Check band identity carefully given the reported 50 kDa band and 40 kDa expectation.
Source: BosterBio CXCR1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P25024.
- Human Protein Atlas. CXCR1 tissue expression.
- PMC5601684 — target-verified WB comparison
- PMC3058860 — target-verified WB comparison
- PMC10974724 — target-verified WB comparison