XCR1 / Chemokine XC receptor 1 · Western blot design guide

Design a Western Blot for XCR1

Real validated XCR1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-XCR1 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

Western blot protocol sheet for XCR1: expected band ~38.5 kDa, hero antibody A04185-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **XCR1** Western blot protocol sheet — expected band ~38.5 kDa, antibody A04185-1, controls and PMC citations. Open the full XCR1 WB guide →

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XCR1 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	~38.5 kDa
	Observed band	~42 kDa
	Gel	5–20% (catalog A04185-1)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	—
	Caveat	Band identity controls
	Gene-set association	MSigDB C7 membership
	Isoform	1 isoform(s)

Section 1

Real Curated XCR1 Western Blot Protocols

The A04185-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	human RT4, human U-87MG, human HEL (catalog A04185-1)
Gel %	5–20% (catalog A04185-1)
Load	30 ug; reducing conditions (catalog A04185-1)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04185-1)
Membrane	nitrocellulose membrane (catalog A04185-1)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog A04185-1)
Primary antibody	A04185-1 · 0.5 μg/mL (catalog A04185-1)
Primary incubation	overnight at 4°C (catalog A04185-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A04185-1)
Secondary incubation	1.5 hour at RT (catalog A04185-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A04185-1)
Detection	ECL (catalog A04185-1)

Section 2

What Is the Expected XCR1 Western Blot Band Size?

XCR1 is predicted at 38.5 kDa and observed near 42 kDa in antibody QC; the cause of the difference is not established.

What am I looking at on my blot?
Band at approximately 42 kDa	Empirical XCR1 band reported in reducing whole-cell lysates; confirm identity with appropriate controls.
Band near 38.5 kDa	Compatible with the UniProt predicted mass, but band identity still requires confirmation.
Weak band in whole-cell lysate	A membrane-localized receptor may be underrepresented in the extracted sample.
Stronger band in a membrane-enriched fraction	Consistent with XCR1 cell-membrane localization.

💡Expected XCR1 appearanceUniProt predicts XCR1 at 38.5 kDa, while antibody QC reports a band at approximately 42 kDa; the cause of the difference is unestablished, so confirm identity with appropriate controls.

How each factor affects band size
UniProt predicted molecular weight	Places the unmodified 333-residue sequence near 38.5 kDa.
Empirical comparison with predicted mass	The reported 42 kDa band runs above the prediction, without an established cause.
Multi-pass membrane localization	May complicate extraction and apparent migration; no specific size shift is established.
Intrachain disulfide between residues 102 and 175	Does not support a doubled-mass band; any effect of reduction on migration is unestablished.

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Membrane-localized XCR1 may be poorly recovered in the lysate.	Check membrane extraction and compare with a membrane-enriched fraction.
Band higher than expected	The reported 42 kDa band exceeds the 38.5 kDa prediction; its cause is unestablished.	Compare with the reported band and confirm identity using XCR1 depletion or an independent antibody.
Band lower than expected	The supplied features do not establish a smaller XCR1 product.	Check sample integrity and confirm the band with XCR1 depletion or an independent antibody.
Multiple bands	Additional bands are not explained by documented XCR1 isoforms.	Use XCR1 depletion or an independent antibody to identify the specific band.
Weak or no signal	Recovery of this multi-pass membrane receptor may be low.	Optimize membrane solubilization and verify protein transfer.

Source: UniProt P46094 Human Protein Atlas product datasheet (A04185-1)

Experiment evidence

Western blot images showing XCR1 band

A04185-1 · XCR1

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Real WB data behind the band-size expectations in this section.

Sample controls for XCR1 Western blot

🧪For positive controls for XCR1 in Western blot, you can use an independently validated XCR1-positive sample, since the supplied HPA evidence identifies 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: XCR1 is a multi-pass membrane protein, and the supplied HPA evidence provides no tissue controls, so sample selection requires independent validation.

HPA tissue expression evidence for XCR1

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 XCR1 Western Blot Tips

Deeper troubleshooting and optimisation questions for XCR1, answered from its protein features.

How should XCR1 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 another XCR1 isoform explain an extra band?

Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. An extra band cannot be assigned to a documented XCR1 isoform from these features alone.

Do annotated modifications prove a band shift?

PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.

Does this guide establish induction of XCR1?

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 XCR1 Western blot?

Transfer · XCR1 is a 333-residue, multi-pass cell-membrane protein with a predicted mass of 38.5 kDa. Check transfer efficiency around the observed 42 kDa band by inspecting the membrane and post-transfer gel. The supplied features do not establish whether wet or semi-dry transfer works better.

How should blocking conditions be optimised for XCR1?

Blocking · Compare blocking conditions using the specific band near 42 kDa and background signal as your readouts. XCR1's membrane location identifies the target, but its listed features do not specify a preferred blocking reagent.

How should the XCR1 band be quantified?

Quantitation · Quantify the band near the supplied observed position of 42 kDa using the same band definition across samples. Keep sample preparation and detection conditions consistent; the predicted mass alone does not identify every detected band as XCR1.

Why might XCR1 run near 42 kDa rather than 38.5 kDa?

Interpretation · XCR1 has a predicted mass of 38.5 kDa, while the supplied observed band is about 42 kDa. Its multi-pass membrane structure may affect migration, but the listed features do not establish the cause of this difference. Do not assign the shift to glycosylation or another modification without evidence.

UniProt lists one disulfide bond for XCR1. If comparing reducing and nonreducing preparations, interpret any mobility difference in light of that feature, but do not assume the bond produces a visible shift; no bond coordinates or shift are supplied.

Use the approximately 42 kDa observed band and 38.5 kDa predicted mass as reference points. The record lists one isoform, one disulfide bond, and no annotated glycosylation or modified residues. Those features alone cannot identify the source of additional bands; verify band identity experimentally.

Boster reagents

XCR1 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

Western blot analysis of CCXCR1/XCR1 using anti-CCXCR1/XCR1 antibody (A04185-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human RT4 whole cell lysates, Lane 2: human U-87MG whole cell lysates, Lane 3: human HEL whole cell lysates, Lane 4: human U20S whole cell lysates, Lane 5: rat spleen tissue lysates, Lane 6: mouse spleen tissue lysates, Lane 7: mouse thymus tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CCXCR1/XCR1 antigen affinity purified polyclonal antibody (Catalog # A04185-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CCXCR1/XCR1 at approximately 42 kDa. The expected band size for CCXCR1/XCR1 is at 39 kDa.

Anti-CCXCR1/XCR1 Antibody Picoband®

Cat # A04185-1

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The catalog reports one anti-XCR1 antibody for Western blot, A04185-1, with stated human, mouse, and rat reactivity. Its WB image shows a band near 42 kDa versus an expected 39 kDa in the specimens tested; no publication evidence is supplied.

Which to pick: A04185-1 is the only listed option. Its WB image includes human cell lysates, rat spleen, and mouse spleen and thymus. Use those tested contexts and the reported 0.5 μg/mL antibody concentration to judge its fit for your samples.

Source: BosterBio XCR1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry P46094.
Human Protein Atlas. XCR1 tissue expression.
PMC9814969 — target-verified WB comparison
PMC7704520 — target-verified WB comparison
PMC11809430 — target-verified WB comparison