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- Table of Contents
JARID2 / Protein Jumonji · Western blot design guide
Design a Western Blot for JARID2
Real validated JARID2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-JARID2 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
JARID2 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 | ~138.7 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated JARID2 Western Blot Protocols
The M01969 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | NCCIT cell lysate (catalog M01969) |
| 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% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M01969; 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 JARID2 Western Blot Band Size?
JARID2 has a predicted mass of 138.7 kDa; isoforms could affect band patterns, but no empirical migration or isoform separation is supplied.
| Band near 138.7 kDa | Consistent with the predicted JARID2 mass; identity requires validation |
| Single band near 138.7 kDa | Isoforms may be unresolved or only one may be detected |
| Several bands at different positions | Could reflect isoforms 1, 2, and 3; their migration is unverified |
| Stronger band in nuclear extract than whole-cell lysate | Consistent with JARID2 nuclear localization |
💡Expected JARID2 appearanceUniProt predicts 138.7 kDa for JARID2, but no empirical band size is supplied; validate any candidate band with antibody specificity controls and nuclear enrichment.
| UniProt predicted mass | Sets a 138.7 kDa reference, not a measured band position |
| Splice isoform 1 | May migrate differently from other isoforms; its mass is unspecified |
| Splice isoform 2 | May migrate differently from other isoforms; its mass is unspecified |
| Splice isoform 3 | May migrate differently from other isoforms; its mass is unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear JARID2 may be poorly represented in the sample | Check nuclear extraction and compare with a nuclear-enriched fraction |
| Band higher than expected | Its identity and migration cannot be established from predicted mass alone | Compare nuclear-enriched samples and verify antibody specificity |
| Band lower than expected | An alternatively spliced isoform is possible, but its size is unknown | Check isoform expression and verify band identity |
| Multiple bands | Isoforms 1, 2, and 3 are annotated; their separation is unverified | Use isoform-aware controls and check antibody specificity |
| Weak or no signal | Nuclear localization may limit signal in whole-cell lysate | Enrich the nuclear fraction and confirm sample loading |
Experiment evidence
Western blot images showing JARID2 band
M01969 · JARID2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for JARID2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for JARID2 in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: No tissue is listed as undetected, so a JARID2 knockdown or knockout line is the clearest negative control.
HPA tissue expression evidence for JARID2
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Cervix | 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 | Low | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Low | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | adipocytes | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced JARID2 Western Blot Tips
Deeper troubleshooting and optimisation questions for JARID2, answered from its protein features.
How should JARID2 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.
How could JARID2 isoforms affect band interpretation?
Isoforms · Three isoforms are listed. Isoform 3 lacks canonical residues 1–172; isoform 2 replaces residues 1–60 and 949–1246. Consider these sequence differences when assessing additional bands, and check whether the antibody target lies in a changed region.
Which JARID2 modifications matter when assessing band shifts?
PTM · The supplied canonical coordinates list phosphoserine at 78 and 455 and N6-acetyllysine at 378. Residue 78 is absent from isoform 3, and isoform 2 replaces the region containing it. These features do not by themselves demonstrate a visible band shift.
Does this guide establish induction of JARID2?
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 JARID2 Western blot?
Transfer · For canonical JARID2 at a predicted 138.7 kDa, choose transfer conditions suited to a large protein. Check the high-mass marker range and whether protein remains in the gel after transfer; adjust the transfer conditions if recovery is poor.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01969 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 JARID2 Western blot signals be quantified?
Quantitation · JARID2 is nuclear and colocalizes with PRC2 on chromatin. Use comparable nuclear or chromatin-containing preparations across samples and quantify the same defined band or isoform consistently. Antibody recognition may depend on the sequence region targeted.
Should JARID2 migrate at its predicted 138.7 kDa?
Interpretation · The canonical sequence has a predicted mass of 138.7 kDa, but no observed band position is supplied. Use 138.7 kDa as a reference, not an expected apparent mass. The listed modifications alone do not establish a visible shift.
Compare each band with the three listed isoforms and the antibody target region. Isoform 3 lacks residues 1–172, while isoform 2 changes both terminal regions. The supplied features cannot establish that any particular unexpected band is an isoform or a modification.
Compare each band with the three listed isoforms and the antibody target region. Isoform 3 lacks residues 1–172, while isoform 2 changes both terminal regions. The supplied features cannot establish that any particular unexpected band is an isoform or a modification.
Boster reagents
JARID2 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-Jarid2 Rabbit Monoclonal Antibody
Cat # M01969
The catalog reports M01969, an anti-Jarid2 rabbit monoclonal antibody with stated human reactivity. Its Western blot image shows Jarid2 expression in NCCIT cell lysate; the supplied evidence covers that tested context only.
Which to pick: M01969 is the only listed JARID2 antibody. It has a Western blot image using NCCIT cell lysate and is listed as reactive with human samples. Confirm suitability for your sample and conditions.
Source: BosterBio JARID2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92833.
- Human Protein Atlas. JARID2 tissue expression.
- PMC7841153 — target-verified WB comparison
- PMC6097338 — target-verified WB comparison
- PMC5130007 — target-verified WB comparison