This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
JAK2 / Tyrosine-protein kinase JAK2 · Western blot design guide
Design a Western Blot for JAK2
Real validated JAK2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-JAK2 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
JAK2 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 | ~130.7 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Colon (total-target IHC; phospho state unverified) +4 more | |
| Negative control | Caudate (total-target IHC; phospho state unverified) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Activation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated JAK2 Western Blot Protocols
The MP00027 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysates treated with Pervanadate (catalog MP00027) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% BSA in TBST (standard starting point) |
| Primary antibody | MP00027; 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 JAK2 Western Blot Band Size?
Full-length JAK2 is predicted at 130.7 kDa; phosphorylation could affect migration, but no empirical band or visible shift is established.
| Band near 130.7 kDa | Consistent with full-length JAK2; confirm identity with appropriate controls. |
| Band near 130.7 kDa after pervanadate treatment | May represent phosphorylated JAK2 recognized by the Y1007 + Y1008 phospho-specific antibody; band identity remains unvalidated here. |
| No band with the phospho-specific antibody | The recognized phosphorylation state may be absent or below detection. |
| Similar band positions before and after phosphorylation | JAK2 phosphorylation does not necessarily produce a visible mobility shift. |
💡Expected JAK2 appearanceUniProt predicts 130.7 kDa for full-length JAK2; no empirical band size is supplied, and the Y1007 + Y1008 phospho-specific antibody requires phosphorylation and band-identity controls.
| UniProt predicted mass | Full-length JAK2 is predicted at 130.7 kDa. |
| Phosphotyrosine 119 | Autocatalytic phosphorylation may affect migration, but no visible shift is established. |
| Phosphoserine 523 | Phosphorylation may affect migration, but no visible shift is established. |
| Phosphotyrosine 868 | Autocatalytic phosphorylation may affect migration, but no visible shift is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| Weak or no signal | The Y1007 + Y1008 antibody recognizes a phosphorylation state that may be scarce. | Compare treated and untreated lysates and check a total JAK2 blot. |
| No band in lysate | JAK2 abundance or the recognized phosphorylation state may be below detection. | Check lysate loading and probe a positive control and total JAK2. |
| Band higher than expected | The supplied features do not establish the cause of a higher band. | Compare with total JAK2 and a positive control; verify band identity. |
| Band lower than expected | The supplied features do not establish a smaller JAK2 product. | Check sample integrity and compare with total JAK2. |
| Multiple bands | The supplied features do not establish distinct JAK2 isoform bands. | Use a positive control and total JAK2 to identify the relevant band. |
Experiment evidence
Western blot images showing JAK2 band
MP00027 · JAK2
M00027 · JAK2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for JAK2 Western blot
HPA tissue expression does not establish phosphorylation at the selected antibody epitope. Verify activation-state controls for the phospho-specific lead.
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for JAK2 in Western blot, you can use colon tissue, where HPA reports high expression.
Positive control: Colon (total-target IHC; phospho state unverified)
Negative control: Caudate (total-target IHC; phospho state unverified)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA reports tissues with high expression and tissues where JAK2 is not detected, making tissue controls feasible.
HPA tissue expression evidence for JAK2
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 | endothelial cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Lung | macrophages | High | Protein (IHC) | HPA → |
| Lymph node | non-germinal center 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 → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Prostate | glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced JAK2 Western Blot Tips
Deeper troubleshooting and optimisation questions for JAK2, answered from its protein features.
Where should the JAK2 band appear?
Band shift · The predicted mass is 130.7 kDa. No empirical apparent band is supplied, so use 130.7 kDa as a reference rather than an exact migration position.
Should I expect multiple JAK2 isoform bands?
Isoforms · The supplied record reports one isoform and lists no isoform names. It therefore provides no basis for assigning additional bands to alternative JAK2 isoforms.
Can phosphorylation explain a shifted JAK2 band?
PTM · UniProt lists 11 modified residues, including phosphoserine 523 and multiple phosphotyrosines. Their presence alone does not establish a visible shift or explain a difference from 130.7 kDa. Compare the band with an appropriate control before assigning a shift to phosphorylation.
UniProt lists phosphotyrosines 119, 372, 373, 570, 813, 868, 966, 972, 1007 and 1008, plus phosphoserine 523. It specifies autocatalysis at tyrosines 119, 868, 966, 972, 1007 and 1008. These are UniProt coordinates; verify the numbering used by the antibody.
Use a total-JAK2 measurement to assess JAK2 abundance and a site-specific phospho measurement to assess modification at the named residue. Report the site using UniProt coordinates and check the antibody’s numbering convention. The listed phosphorylation sites do not establish how much JAK2 is phosphorylated in a sample.
UniProt lists phosphotyrosines 119, 372, 373, 570, 813, 868, 966, 972, 1007 and 1008, plus phosphoserine 523. It specifies autocatalysis at tyrosines 119, 868, 966, 972, 1007 and 1008. These are UniProt coordinates; verify the numbering used by the antibody.
Use a total-JAK2 measurement to assess JAK2 abundance and a site-specific phospho measurement to assess modification at the named residue. Report the site using UniProt coordinates and check the antibody’s numbering convention. The listed phosphorylation sites do not establish how much JAK2 is phosphorylated in a sample.
Does this guide establish induction of JAK2?
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 JAK2?
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 MP00027 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 JAK2 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
How should I interpret unexpected JAK2 bands?
Interpretation · Compare them with the 130.7 kDa predicted mass and verify antibody specificity before assigning an identity. The supplied record lists phosphorylation sites but no empirical band position; it does not establish that phosphorylation causes a visible shift. It also reports only one isoform.
Yes. UniProt places JAK2 in the cytoplasm and nucleus and describes it as a peripheral protein of the endomembrane system. Consider which fractions were collected when comparing samples; a band’s absence from one fraction alone does not establish absence of JAK2 from the sample.
Yes. UniProt places JAK2 in the cytoplasm and nucleus and describes it as a peripheral protein of the endomembrane system. Consider which fractions were collected when comparing samples; a band’s absence from one fraction alone does not establish absence of JAK2 from the sample.
Boster reagents
JAK2 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-Phospho-JAK2 (Y1007 + Y1008) Monoclonal Antibody
Cat # MP00027
Real WB data
Anti-JAK2 Monoclonal Antibody
Cat # M00027
Two the supplier JAK2 antibodies have supplied Western blot image evidence: MP00027 for JAK2 phosphorylation in pervanadate-treated Jurkat lysates, and M00027 for JAK2 detection. The supplied records do not establish broader validation or performance across sample types.
Which to pick: Choose MP00027 for the pictured phosphorylation assay in pervanadate-treated Jurkat lysates, or M00027 for the pictured JAK2 Western blot. Both have WB images; reactivity beyond the described samples is not specified.
Source: BosterBio JAK2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O60674.
- Human Protein Atlas. JAK2 tissue expression.
- PMC5256931 — target-verified WB comparison
- PMC11291247 — target-verified WB comparison
- PMC3319112 — target-verified WB comparison