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- Table of Contents
YTHDC2 / 3'-5' RNA helicase YTHDC2 · Western blot design guide
Design a Western Blot for YTHDC2
Real validated YTHDC2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-YTHDC2 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
YTHDC2 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 | ~160.2 kDa | |
| Observed band | ~175 kDa | |
| Gel | 5–20% (catalog A09692-1) | |
| Positive control | Colon (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated YTHDC2 Western Blot Protocols
The A09692-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Daudi, human Hela, human 293T (catalog A09692-1) |
| Gel % | 5–20% (catalog A09692-1) |
| Load | 30 ug; reducing conditions (catalog A09692-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A09692-1) |
| Membrane | nitrocellulose membrane (catalog A09692-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A09692-1) |
| Primary antibody | A09692-1 · 0.5 μg/mL (catalog A09692-1) |
| Primary incubation | overnight at 4°C (catalog A09692-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A09692-1) |
| Secondary incubation | 1.5 hour at RT (catalog A09692-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A09692-1) |
| Detection | ECL (catalog A09692-1) |
Section 2
What Is the Expected YTHDC2 Western Blot Band Size?
YTHDC2 is predicted at 160.2 kDa but observed near 175 kDa on reducing Western blots; the cause of this difference is not established.
| Band near 175 kDa | empirical YTHDC2 band in reducing whole-cell lysates; confirm identity with controls |
| Band near 160 kDa | close to the 160.2 kDa sequence-predicted mass; identity requires confirmation |
| Close doublet near the main band | could reflect phosphostates at listed serines, but distinct migration is unproven |
| Little signal in a cytoplasm-depleted sample | consistent with YTHDC2's cytoplasmic location |
💡Expected YTHDC2 appearanceYTHDC2 has a predicted mass of 160.2 kDa and an empirical band near 175 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm band identity with controls.
| Predicted sequence mass | 160.2 kDa calculated; the empirical band is near 175 kDa for an unestablished reason |
| Phosphoserines 1089, 1090, and 1092 | may affect migration; no visible shift is established |
| Phosphoserine 1202 | may affect migration; no visible shift is established |
| Phosphoserines 1263, 1267, and 1281 | may affect migration; no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | insufficient cytoplasmic YTHDC2 in the sample or inadequate detection | check cytoplasmic sample content and a positive-control lysate |
| Band higher than expected | the empirical YTHDC2 band is near 175 kDa despite a 160.2 kDa predicted mass; the cause is unestablished | compare with a positive-control lysate and verify antibody specificity |
| Band lower than expected | possible degradation or unrelated antibody binding; no cleavage feature is listed | check sample integrity and verify the band with an independent antibody or YTHDC2 depletion |
| Multiple bands | possible phosphostates or unrelated antibody binding; distinct migration is unproven | compare phosphatase-treated samples and verify bands with YTHDC2 depletion |
| Fragments below expected size | possible sample degradation; no programmed cleavage is listed | prepare fresh lysate with protease inhibitors and check fragment specificity |
Experiment evidence
Western blot images showing YTHDC2 band
A09692-1 · YTHDC2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for YTHDC2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for YTHDC2 in Western blot, you can use colon tissue lysate.
Positive control: Colon (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: Cytoplasmic localization supports tissue lysate testing, with adipose tissue as an HPA not-detected negative control.
HPA tissue expression evidence for YTHDC2
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 |
|---|---|---|---|---|
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | High | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | High | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced YTHDC2 Western Blot Tips
Deeper troubleshooting and optimisation questions for YTHDC2, answered from its protein features.
How should YTHDC2 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.
Are multiple YTHDC2 isoforms expected?
Isoforms · The supplied entry lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands. Investigate additional bands with a YTHDC2-specific control before assigning them to YTHDC2.
Could phosphorylation shift the YTHDC2 band?
PTM · UniProt lists phosphoserines at positions 1089, 1090, 1092, 1202, 1263, 1267 and 1281, using UniProt numbering. These sites do not establish a visible shift or explain the 175 kDa band. Compare phosphatase-treated and untreated samples to test whether phosphorylation affects mobility.
Does this guide establish induction of YTHDC2?
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 YTHDC2 Western blot?
Transfer · YTHDC2 is a large protein with a predicted mass of 160.2 kDa. Choose transfer conditions suitable for proteins in this size range, and check transfer near 160–175 kDa using the ladder and the gel after transfer. The supplied features do not specify a particular membrane or transfer setting.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A09692-1 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 YTHDC2 bands be quantified?
Quantitation · Measure the same confirmed YTHDC2 band across comparable samples and normalize for sample loading. If studying its listed phosphoserine sites, assess phosphorylation separately from total YTHDC2 abundance; a mobility change alone does not quantify phosphorylation.
Why might YTHDC2 appear near 175 kDa?
Interpretation · YTHDC2 has a predicted mass of 160.2 kDa, while the supplied Western blot observation is approximately 175 kDa. The listed features do not establish the cause of that difference. Check the band against a molecular weight ladder and confirm its identity with a YTHDC2-specific control.
YTHDC2 is annotated with meiosis, spermatogenesis and oogenesis, and is located in the cytoplasm, including the perinuclear region. These features support testing relevant meiotic or reproductive samples and retaining the cytoplasmic material during preparation. No specific inducing treatment is supplied.
The entry lists one isoform, seven phosphoserines, and no signal peptide, propeptide or glycosylation sites. None of these features establishes the identity of an extra band. Confirm whether each band is YTHDC2 before attributing it to processing or phosphorylation.
YTHDC2 is annotated with meiosis, spermatogenesis and oogenesis, and is located in the cytoplasm, including the perinuclear region. These features support testing relevant meiotic or reproductive samples and retaining the cytoplasmic material during preparation. No specific inducing treatment is supplied.
The entry lists one isoform, seven phosphoserines, and no signal peptide, propeptide or glycosylation sites. None of these features establishes the identity of an extra band. Confirm whether each band is YTHDC2 before attributing it to processing or phosphorylation.
Boster reagents
YTHDC2 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-YTHDC2 Antibody Picoband®
Cat # A09692-1
The listed anti-YTHDC2 antibody, A09692-1, has a Western blot image using human, rat, and mouse cell lysates. Its reported band is approximately 175 kDa versus an expected 160 kDa. No independent validation is supplied.
Which to pick: A09692-1 is the only listed option. Its Western blot image uses human Daudi, HeLa, and 293T; rat C6; and mouse NIH/3T3 whole cell lysates, with 30 µg per lane and antibody at 0.5 µg/mL. Match your sample and conditions to these reported contexts.
Source: BosterBio YTHDC2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9H6S0.
- Human Protein Atlas. YTHDC2 tissue expression.
- PMC12775497 — target-verified WB comparison
- PMC10457424 — target-verified WB comparison
- PMC8681016 — target-verified WB comparison