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- Table of Contents
ZFP36L2 / mRNA decay activator protein ZFP36L2 · Western blot design guide
Design a Western Blot for ZFP36L2
Source-linked ZFP36L2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ZFP36L2 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
ZFP36L2 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 | ~51.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Epididymis (IHC candidate; verify WB) +4 more | |
| Negative control | Caudate (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
Source-Linked ZFP36L2 Western Blot Protocol Options
The A05565-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | PC12 (40ug), HCT116 (40ug), CT26 (40ug), A375 (40ug), Myla2059 (40ug) (catalog A05565-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A05565-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected ZFP36L2 Western Blot Band Size?
ZFP36L2 is predicted at 51.1 kDa; phosphorylation could affect migration, but no empirical band or demonstrated shift is supplied.
| Band near 51.1 kDa | consistent with the predicted size of ZFP36L2; confirm identity with a specific control |
| Band slightly above 51.1 kDa | phosphorylation could affect migration, but a shift is not established |
| Close doublet near 51.1 kDa | different phosphorylation states are possible, but distinct bands are not established |
| Several closely spaced bands near 51.1 kDa | could reflect phosphorylation states; verify that the bands are ZFP36L2 |
💡Expected ZFP36L2 appearanceUniProt predicts 51.1 kDa, but no empirical band size is supplied; use a ZFP36L2-specific control to identify the band because its listed phosphorylation sites do not establish a migration shift.
| Predicted molecular weight | sets a 51.1 kDa reference for the full-length protein |
| Phosphoserine 57 and 125 | phosphorylation could alter migration, with no demonstrated shift |
| Phosphothreonine 238 | phosphorylation could alter migration, with no demonstrated shift |
| RPS6KA1-dependent phosphoserine 490 and 492 | phosphorylation could alter migration, with no demonstrated shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ZFP36L2 may be missed during preparation of its nuclear or cytoplasmic fractions | check both fractions and include a positive control |
| Band higher than expected | phosphorylation is possible, but the band's identity and shift are unverified | confirm identity with a specific control and compare phosphatase-treated material |
| Band lower than expected | the band may be a fragment or nonspecific signal | confirm identity with a specific control and check sample integrity |
| Multiple bands | phosphorylation states are possible, but distinct migration is unverified | compare phosphatase-treated material and use a specific identity control |
| Weak or no signal | expression or recovery of nuclear and cytoplasmic ZFP36L2 may be low | check fraction recovery and include a positive control |
Experiment evidence
Western blot images showing ZFP36L2 band
A05565-1 · ZFP36L2
A05565 · ZFP36L2
A05565 · ZFP36L2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ZFP36L2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ZFP36L2 in Western blot, you can use epididymis lysate.
Positive control: Epididymis (IHC candidate; verify WB)
Negative control: Caudate (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: ZFP36L2 is intracellular, and HPA identifies caudate as a candidate negative tissue; confirm its absence by Western blot.
HPA tissue expression evidence for ZFP36L2
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 |
|---|---|---|---|---|
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | High | Protein (IHC) | HPA → |
| Small intestine | glandular cells | High | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | High | Protein (IHC) | HPA → |
| Tonsil | non-germinal center cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ZFP36L2 Western Blot Tips
Deeper troubleshooting and optimisation questions for ZFP36L2, answered from its protein features.
Where should the main ZFP36L2 band appear?
Band shift · The predicted mass is 51.1 kDa. Use that as a reference when examining the blot; no empirical apparent band position is supplied.
Are alternative isoforms expected to create multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no specific isoform explanation for multiple bands; investigate other causes before assigning them to ZFP36L2 isoforms.
Could phosphorylation change ZFP36L2 band migration?
PTM · UniProt lists phosphoserine at positions 57, 125, 490, and 492, and phosphothreonine at 238. Positions 490 and 492 are attributed to RPS6KA1. These are UniProt coordinates, which may differ from antibody or paper numbering. The listed sites alone do not establish a visible shift.
Does this guide establish induction of ZFP36L2?
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 ZFP36L2?
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 A05565-1 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 ZFP36L2 abundance be quantified?
Quantitation · Quantify a consistently identified band across comparable samples. Because ZFP36L2 occurs in both nucleus and cytoplasm, keep the sampled fraction consistent; a change in one fraction may reflect altered distribution rather than total abundance.
How should RPS6KA1-related bands be assessed?
Interpretation · RPS6KA1 is listed as phosphorylating ZFP36L2 at serines 490 and 492 in UniProt coordinates. Compare band patterns across samples with relevant RPS6KA1 activity, while avoiding attribution of a band shift to those sites without further evidence.
ZFP36L2 is reported in both nucleus and cytoplasm and shuttles between them in an XPO1/CRM1-dependent manner. Examine both fractions when assessing localization, and compare matched fractions when interpreting differences in band intensity.
Compare them with the 51.1 kDa predicted mass and check whether they occur in nuclear or cytoplasmic fractions. The record lists phosphorylation sites but no observed band position or alternative sequence, so neither phosphorylation nor isoforms can be assigned as the cause from these features alone.
ZFP36L2 is reported in both nucleus and cytoplasm and shuttles between them in an XPO1/CRM1-dependent manner. Examine both fractions when assessing localization, and compare matched fractions when interpreting differences in band intensity.
Compare them with the 51.1 kDa predicted mass and check whether they occur in nuclear or cytoplasmic fractions. The record lists phosphorylation sites but no observed band position or alternative sequence, so neither phosphorylation nor isoforms can be assigned as the cause from these features alone.
Boster reagents
ZFP36L2 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-TIS11D (H243) ZFP36L2 Antibody
Cat # A05565-1
Real WB data
Anti-TIS11D ZFP36L2 Antibody
Cat # A05565
Two the supplier anti-ZFP36L2 antibodies have WB images. A05565-1 shows testing at 1:500 with PC12, HCT116, CT26, A375, and Myla2059 lysates. A05565’s caption says only ‘various cells’ and gives a secondary antibody dilution; it provides less detail about the tested samples.
Which to pick: Choose A05565-1 if you need listed rat reactivity or a WB example with named lysates and a primary antibody dilution. It lists human, mouse, and rat; A05565 lists human and mouse. Both have WB images, but A05565’s caption does not name its samples.
Source: BosterBio ZFP36L2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P47974.
- Human Protein Atlas. ZFP36L2 tissue expression.
- PMC10432607 — target-verified WB comparison
- PMC9334557 — target-verified WB comparison
- PMC4022657 — target-verified WB comparison