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- Table of Contents
HRK / Activator of apoptosis harakiri · Western blot design guide
Design a Western Blot for HRK
Real validated HRK Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HRK 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
HRK 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 | ~9.9 kDa | |
| Gel | 15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | No observed band reported | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated HRK Western Blot Protocols
The A09125-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A09125-1; 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 HRK Western Blot Band Size?
HRK’s predicted size is 9.9 kDa; membrane localization may affect recovery, but empirical migration and any size shift are unestablished.
| Band near 9.9 kDa | consistent with predicted HRK size, pending identity controls |
| Band in a membrane fraction | consistent with HRK membrane localization |
| Band in a mitochondrial fraction | consistent with HRK mitochondrial localization |
| Faint or absent band in soluble lysate | HRK may remain in the membrane fraction |
💡Expected HRK appearanceHRK has a predicted mass of 9.9 kDa, but no empirical band size is supplied; confirm the identity of any band near that size.
| UniProt predicted mass of 9.9 kDa | provides the reference size for a putative HRK band |
| UniProt molecular weight of 9884 Da | expresses the same predicted sequence mass in daltons |
| Predicted monomer mass | supports looking near 9.9 kDa without establishing apparent migration |
| Sequence-based mass estimate | does not establish an experimentally observed band position |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated HRK may be poorly recovered | Check membrane or mitochondrial fractions and confirm protein recovery |
| Band higher than expected | Band identity or apparent migration is unverified | Test HRK depletion and an independent antibody |
| Band lower than expected | The band may be a fragment or nonspecific signal | Compare fresh samples and test HRK depletion |
| Multiple bands | Their identities are unverified; UniProt lists one HRK isoform | Use HRK depletion to identify the specific band |
| Weak or no signal | HRK abundance or membrane protein recovery may be low | Check loading, membrane recovery, and antibody performance |
Experiment evidence
Western blot images showing HRK band
A09125-1 · HRK
A09125 · HRK
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HRK Western blot
🧪For positive controls for HRK in Western blot, you can use no tissue or cell line identified by the supplied HPA evidence.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA supplies no tissue data, so a positive tissue control cannot be selected.
HPA tissue expression evidence for HRK
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced HRK Western Blot Tips
Deeper troubleshooting and optimisation questions for HRK, answered from its protein features.
Where should the HRK band appear relative to its predicted mass?
Band shift · HRK is 91 amino acids with a predicted mass of 9.9 kDa. No observed band position is supplied, so use 9.9 kDa as a reference rather than an expected apparent position. Confirm candidate bands with an HRK-specific control.
Could an HRK isoform explain a second band?
Isoforms · The supplied features list one isoform and no alternative sequence. They do not support assigning a second band to another HRK isoform; verify its identity before interpreting it.
Do listed modifications explain an HRK band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features therefore provide no specific modification to test as the cause of a shifted band. They also do not establish why an apparent mass might differ from 9.9 kDa.
Does this guide establish induction of HRK?
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 HRK Western blot?
Transfer · HRK is a small, 9.9 kDa single-pass membrane protein. Choose a transfer setup intended to retain small proteins, and check HRK retention when optimizing transfer conditions. Its membrane association also makes recovery during sample preparation worth checking.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A09125-1 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 HRK abundance be quantified?
Quantitation · Quantify a verified HRK band within the assay's linear range. Because HRK is mitochondrial and membrane-associated, keep sample preparation consistent across conditions and confirm comparable recovery before interpreting intensity differences as changes in abundance.
How should unexpected higher-mass HRK bands be interpreted?
Interpretation · HRK is reported to interact with BCL2, BCL2L1 and C1QBP, but those interactions alone do not identify a higher-mass Western blot band. The supplied features give no observed band position. Verify band identity and assess sample preparation before assigning an explanation.
Boster reagents
HRK 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-HRK Antibody
Cat # A09125-1
Real WB data
Anti-Hrk Antibody
Cat # A09125
Two the supplier anti-HRK antibodies have Western blot images: A09125-1 with SH-SY5Y cell lysate at 1:1000, and A09125 with mouse pancreas lysate at 2.5 and 5 μg/mL. These examples document specific test conditions, not broad validation across listed species.
Which to pick: For rat samples, A09125-1 is the only antibody with rat reactivity listed, though its shown blot uses SH-SY5Y cells. For mouse pancreas, A09125 has the directly relevant blot. Both list human and mouse reactivity.
Source: BosterBio HRK gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O00198.
- Human Protein Atlas. HRK tissue expression.
- PMC3069046 — target-verified WB comparison
- PMC5797550 — target-verified WB comparison