CD1A / T-cell surface glycoprotein CD1a · Western blot design guide

Design a Western Blot for CD1A

Real validated CD1A Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CD1A 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 CD1A: expected band ~37.1 kDa, hero antibody PA1875, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **CD1A** Western blot protocol sheet — expected band ~37.1 kDa, antibody PA1875, controls and PMC citations. Open the full CD1A WB guide →

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CD1A 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	~37.1 kDa
	Observed band	~37 kDa
	Gel	5–20% (catalog PA1875)
	Positive control ⓘ	Skin (IHC candidate; verify WB) +2 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	Glycosylation-state controls
	Gene-set association	MSigDB C7 membership
	Isoform	1 isoform(s)

Section 1

Real Curated CD1A Western Blot Protocols

The PA1875 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 Jurkat (catalog PA1875)
Gel %	5–20% (catalog PA1875)
Load	30 ug; reducing conditions (catalog PA1875)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog PA1875)
Membrane	nitrocellulose membrane (catalog PA1875)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog PA1875)
Primary antibody	PA1875 · 0.5 μg/mL (catalog PA1875)
Primary incubation	overnight at 4°C (catalog PA1875)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog PA1875)
Secondary incubation	1.5 hour at RT (catalog PA1875)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog PA1875)
Detection	ECL (catalog PA1875)

Section 2

What Is the Expected CD1A Western Blot Band Size?

CD1a is predicted at 37.1 kDa and observed at ~37 kDa; the cause of their small apparent difference is not established.

What am I looking at on my blot?
Band at ~37 kDa	Empirical CD1a band near its 37.1 kDa predicted precursor mass
Band below ~37 kDa	Could reflect signal peptide removal; identity requires confirmation
Band above ~37 kDa	Could reflect N-linked glycosylation; a visible shift is unproven
Broad smear near the expected region	Could reflect variable N-linked glycosylation; heterogeneity is unproven

💡Expected CD1A appearanceCD1a has a predicted precursor mass of 37.1 kDa and an empirical band at ~37 kDa; confirm band identity with a positive control and a second antibody.

How each factor affects band size
Predicted precursor mass	37.1 kDa predicted; the empirical CD1a band is ~37 kDa
N-linked glycosylation site at Asn37	Could affect migration if occupied; shift magnitude is unknown
N-linked glycosylation site at Asn60	Could affect migration if occupied; shift magnitude is unknown
N-linked glycosylation site at Asn74	Could affect migration if occupied; shift magnitude is unknown
N-linked glycosylation site at Asn145	Could affect migration if occupied; shift magnitude is unknown
Signal peptide at residues 1–16	Cleavage yields a smaller mature chain, without a demonstrated resolvable shift

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Membrane-associated CD1a may be poorly recovered	Check membrane extraction and run a CD1a-positive control
Band higher than expected	N-linked glycosylation could affect migration	Compare paired samples with and without N-glycan removal
Band lower than expected	Signal peptide cleavage could reduce chain size	Verify identity with a second antibody and a positive control
Broad smear instead of sharp band	Variable N-linked glycosylation is possible	Compare paired samples with and without N-glycan removal
Multiple bands	Glycosylation or signal peptide processing could contribute	Compare glycan removal results and confirm bands with a second antibody
Weak or no signal	Membrane-associated CD1a may be underrepresented in the preparation	Enrich membrane protein and check a CD1a-positive control

Source: UniProt P06126 Human Protein Atlas product datasheet (PA1875)

Experiment evidence

Western blot images showing CD1A band

PA1875 · CD1A

M00375-2 · CD1A

← swipe / scroll for more images →

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

Sample controls for CD1A Western blot

🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CD1A in Western blot, you can use skin tissue, where HPA reports high expression.

Positive control: Skin (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: CD1A is membrane-associated, so whole-tissue lysate may dilute its signal.

HPA tissue expression evidence for CD1A

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
Skin	fibrohistiocytic cells	High	Protein (IHC)	HPA →
Thymus	cortical cells	High	Protein (IHC)	HPA →
Tonsil	non-germinal center 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 →
Adrenal gland	glandular cells	Not detected	Protein (IHC)	HPA →
Appendix	glandular cells	Not detected	Protein (IHC)	HPA →
Bone marrow	hematopoietic cells	Not detected	Protein (IHC)	HPA →
Breast	adipocytes	Not detected	Protein (IHC)	HPA →

Section 3

Advanced CD1A Western Blot Tips

Deeper troubleshooting and optimisation questions for CD1A, answered from its protein features.

How should CD1A 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 CD1A isoform explain an extra band?

Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning an extra band to a CD1A splice isoform.

Which CD1A glycosylation sites matter when comparing bands?

PTM · UniProt lists N-linked sites at Asn37, Asn60, Asn74 and Asn145. Keep these UniProt coordinates when comparing with antibody or paper numbering, which may use a different convention. The listed sites do not establish how much glycan is present in your sample or whether a band shift will be visible.

Does this guide establish induction of CD1A?

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 CD1A Western blot?

Transfer · CD1A is a single-pass membrane protein with a reported band near 37 kDa. Check recovery of the membrane fraction and transfer at that size. The supplied features do not establish a preferred transfer method.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the PA1875 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 CD1A bands be quantified?

Quantitation · Quantify the same consistently identified CD1A band across samples. The reported band is about 37 kDa, but four N-linked sites could complicate band assignment if additional forms appear. CD1A is found at the cell and endosome membranes, so use comparable sample preparation across lanes.

Why might CD1A migrate differently from its predicted mass?

Interpretation · The predicted mass is 37.1 kDa, and the reported band is about 37 kDa. CD1A has a signal peptide at UniProt residues 1–16 and four N-linked glycosylation sites. These features can affect the mature protein, but their presence alone does not establish a visible shift or explain any measured mass difference.

UniProt lists two disulfide bonds. Compare bands only under consistent reducing and denaturing conditions; the feature count alone does not predict a particular mobility change or extra band.

Compare them with the reported ~37 kDa band, then consider the listed signal peptide, N-linked sites and disulfide bonds. CD1A also forms a heterodimer with B2M, but these features alone cannot identify an unexpected band. Verify its identity before assigning a specific cause.

Boster reagents

CD1A 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 CD1a using anti-CD1a antibody (PA1875). 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 Jurkat whole cell 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-CD1a antigen affinity purified polyclonal antibody (Catalog # PA1875) 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 CD1a at approximately 37 kDa. The expected band size for CD1a is at 37 kDa.

Anti-CD1a Antibody Picoband®

Cat # PA1875

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Real WB data Western blot analysis of CD1a expression in human fetal thymus lysate.

Anti-CD1a Rabbit Monoclonal Antibody

Cat # M00375-2

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Two the supplier anti-CD1a antibodies are listed for human Western blotting. PA1875 has an image showing an approximately 37 kDa band in Jurkat whole-cell lysate; M00375-2 has an image from human fetal thymus lysate. These images document specific tested samples, not broader validation.

Which to pick: Choose PA1875 for a documented Jurkat lysate protocol with a reported 37 kDa band and 0.5 μg/mL primary antibody. Choose M00375-2 if the fetal thymus example better matches your sample; its supplied caption gives no antibody concentration or band size. Both list human reactivity.

Source: BosterBio CD1A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry P06126.
Human Protein Atlas. CD1A tissue expression.
PMC5347785 — target-verified WB comparison