This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ANGPTL2 / Angiopoietin-related protein 2 · Western blot design guide
Design a Western Blot for ANGPTL2
Source-linked ANGPTL2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ANGPTL2 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
ANGPTL2 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 | ~57.1 kDa | |
| Observed band | ~60–65 kDa | |
| Gel | 10% (catalog A05747-1) | |
| Positive control | Testis (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 + Cleaved | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked ANGPTL2 Western Blot Protocol Options
The A05747-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HUVEC, human SH-SY5Y, rat heart, mouse heart (catalog A05747-1) |
| Gel % | 10% (catalog A05747-1) |
| Load | 30 ug; reducing conditions (catalog A05747-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05747-1) |
| Membrane | nitrocellulose membrane (catalog A05747-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05747-1) |
| Primary antibody | A05747-1 · 0.5 μg/mL (catalog A05747-1) |
| Primary incubation | overnight at 4°C (catalog A05747-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05747-1) |
| Secondary incubation | 1.5 hour at RT (catalog A05747-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05747-1) |
| Detection | ECL (catalog A05747-1) |
Section 2
What Is the Expected ANGPTL2 Western Blot Band Size?
ANGPTL2 has a predicted 57.1 kDa precursor and an empirical ~60–65 kDa band; the cause of their difference is not established.
| Band at ~60–65 kDa | Empirical ANGPTL2 band in reducing lysate blots; confirm identity with antibody controls |
| Band near 57.1 kDa | Near the predicted precursor mass; identity requires confirmation |
| Band below the precursor position | Could reflect signal peptide cleavage; its migration is not established |
| Several bands at different positions | Could include isoforms 1 and 2, if their migration differs |
| Little or no band in whole-cell lysate | ANGPTL2 is secreted and may be enriched in conditioned medium |
💡Expected ANGPTL2 appearanceUniProt predicts a 57.1 kDa precursor, while antibody QC detects ~60–65 kDa in reducing lysate blots; the cause of the difference is unproven, so use ordinary band-identity controls.
| Predicted precursor mass | 57.1 kDa is the sequence-based reference; the empirical band is ~60–65 kDa |
| N-linked glycosylation at Asn164 | May affect apparent mass; its contribution to the observed band is unproven |
| N-linked glycosylation at Asn192 | May affect apparent mass; its contribution to the observed band is unproven |
| Signal peptide at residues 1–22 | Cleavage makes the mature chain smaller than the precursor; migration is not supplied |
| Isoforms 1 and 2 | May differ in size, but their masses and separation on a blot are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Secreted ANGPTL2 may be depleted from cells | Check conditioned medium alongside lysate |
| Band higher than expected | N-linked glycosylation may affect migration, but the cause of ~60–65 kDa is unproven | Compare matched mock-treated and deglycosylated samples with band-identity controls |
| Band lower than expected | Signal peptide cleavage may reduce size relative to the precursor | Compare cellular and secreted fractions and verify band identity |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible at Asn164 and Asn192 | Compare matched mock-treated and deglycosylated samples |
| Multiple bands | Isoforms 1 and 2 may differ in migration; their separation is unknown | Use isoform-aware controls to identify each band |
| Weak or no signal | Secreted ANGPTL2 may be scarce in the sampled lysate | Check conditioned medium and a validated positive control |
Experiment evidence
Western blot images showing ANGPTL2 band
A05747-1 · ANGPTL2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ANGPTL2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ANGPTL2 in Western blot, you can use testis tissue.
Positive control: Testis (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside your samples.
⚠️Feasibility: ANGPTL2 is secreted, so whole-cell lysates may have weak signal; consider conditioned medium.
HPA tissue expression evidence for ANGPTL2
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 |
|---|---|---|---|---|
| Testis | Leydig 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 ANGPTL2 Western Blot Tips
Deeper troubleshooting and optimisation questions for ANGPTL2, answered from its protein features.
Does signal-peptide removal explain ANGPTL2 band size?
Band shift · ANGPTL2 has a signal peptide at canonical residues 1–22 and is annotated as secreted. Consider processing when comparing a detected band with the 57.1 kDa predicted mass, but the supplied features do not establish how much processing changes its apparent migration.
How could ANGPTL2 isoforms affect the blot?
Isoforms · Two isoforms are listed. Isoform 2 lacks residues 1–302 of the canonical sequence, including the annotated signal peptide and both glycosylation sites. Check which isoform an antibody can recognize before assigning a band; the supplied features do not give isoform-specific apparent masses.
Which ANGPTL2 glycosylation sites matter for band interpretation?
PTM · UniProt annotates N-linked glycosylation at Asn164 and Asn192, using canonical sequence coordinates. Record the coordinate convention when comparing antibody or paper numbering. The site annotations alone do not establish a visible band shift.
Does this guide establish induction of ANGPTL2?
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.
How should transfer be checked for ANGPTL2?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05747-1 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 ANGPTL2 bands be quantified across samples?
Quantitation · Compare the same sample type and band definition under consistent blot conditions. ANGPTL2 is secreted, has two isoforms, and has two annotated N-linked glycosylation sites; these features make it useful to report whether quantitation includes one band or several.
Why might ANGPTL2 appear near 60–65 kDa?
Interpretation · The supplied apparent band is 60–65 kDa, versus a predicted 57.1 kDa. ANGPTL2 has a signal peptide at residues 1–22 and two annotated N-linked glycosylation sites. These features are relevant when interpreting migration, but they do not establish the cause or size of the observed difference.
ANGPTL2 is annotated as secreted, so conditioned medium is a relevant sample alongside cell lysate. State which sample was measured when comparing bands or abundance across experiments.
Check antibody recognition of isoforms 1 and 2: isoform 2 lacks canonical residues 1–302. Also consider the annotated signal peptide, N-linked glycosylation sites, and two disulfide bonds when reviewing sample preparation and migration. These features alone cannot identify an unexpected band.
ANGPTL2 is annotated as secreted, so conditioned medium is a relevant sample alongside cell lysate. State which sample was measured when comparing bands or abundance across experiments.
Check antibody recognition of isoforms 1 and 2: isoform 2 lacks canonical residues 1–302. Also consider the annotated signal peptide, N-linked glycosylation sites, and two disulfide bonds when reviewing sample preparation and migration. These features alone cannot identify an unexpected band.
Boster reagents
ANGPTL2 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-ANGPTL2 Antibody Picoband®
Cat # A05747-1
A05747-1 is a rabbit polyclonal anti-ANGPTL2 antibody with a WB image showing a 60–65 kDa band in human cell and rat and mouse heart lysates; the expected size is 57 kDa. No orthogonal validation is supplied.
Which to pick: A05747-1 is the only listed option. Its reported reactivity is human, mouse, and rat; its WB image uses human HUVEC and SH-SY5Y cells and rat and mouse heart tissue. Match your sample to these tested contexts where possible.
Source: BosterBio ANGPTL2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UKU9.
- Human Protein Atlas. ANGPTL2 tissue expression.
- PMC5052800 — target-verified WB comparison
- PMC4317964 — target-verified WB comparison
- PMC9976406 — target-verified WB comparison