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- Table of Contents
BMF / Bcl-2-modifying factor · Western blot design guide
Design a Western Blot for BMF
Real validated BMF Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BMF 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
BMF 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 | ~20.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (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 | — | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated BMF Western Blot Protocols
The A02432-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | 293 cell lysate (catalog A02432-1) |
| Gel % | 12–15% (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 | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A02432-1 · (A) 1 and (B) 2 μg/ml (catalog A02432-1) |
| 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 BMF Western Blot Band Size?
BMF is predicted at 20.5 kDa; three splice isoforms could affect band patterns, but no distinct migration or empirical band size is demonstrated.
| Band near 20.5 kDa | Consistent with the predicted BMF size; confirm identity with controls |
| Several bands at different positions | Could reflect BMF isoforms 1, 2, and 3; band identities require verification |
| One sharp band despite three isoforms | Isoforms need not produce distinct visible bands |
| Band away from 20.5 kDa | An isoform is possible, but its size and the band identity are unestablished |
💡Expected BMF appearanceBMF has a predicted mass of 20.5 kDa, but no empirical band size or isoform migration pattern is supplied; verify any detected band with ordinary identity controls.
| UniProt predicted mass | The listed 184-residue sequence has a calculated mass of 20.5 kDa |
| Splice isoform 1 | Its individual apparent band size is not supplied |
| Splice isoform 2 | Its size may differ, but no individual apparent band size is supplied |
| Splice isoform 3 | Its size may differ, but no individual apparent band size is supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | BMF abundance in this lysate is unknown | Check loading and use a BMF positive control |
| Band higher than expected | An isoform or an unrelated band is possible; no size assignment is supplied | Compare with a BMF positive control and confirm specificity |
| Band lower than expected | An isoform or an unrelated band is possible; no size assignment is supplied | Confirm identity with an independent BMF antibody or BMF depletion |
| Multiple bands | BMF has three named splice isoforms, but their migration is unknown | Test band identities with isoform-specific controls or BMF depletion |
| Weak or no signal | Sample BMF abundance or assay sensitivity may be insufficient | Check loading and detection with a BMF positive control |
Experiment evidence
Western blot images showing BMF band
A02432-1 · BMF
M02432-1 · BMF
A02432 · BMF
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BMF Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for BMF in Western blot, you can use adrenal gland lysate, which HPA rates as high expression.
Positive control: Adrenal gland (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: HPA reports BMF as not detected in adipose tissue, providing a tissue negative control.
HPA tissue expression evidence for BMF
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 → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Lung | macrophages | Low | Protein (IHC) | HPA → |
Section 3
Advanced BMF Western Blot Tips
Deeper troubleshooting and optimisation questions for BMF, answered from its protein features.
What band size is expected for canonical BMF?
Band shift · The supplied predicted mass is 20.5 kDa for the 184-residue canonical sequence. No empirical apparent band size is supplied, so use 20.5 kDa as a reference, not a guaranteed migration position.
Could BMF isoforms produce different bands?
Isoforms · Three isoforms are listed. Isoform 2 lacks UniProt residues 131–151; isoform 3 replaces residues 98–129 and lacks residues 130–184. These sequence differences could affect band size and antibody recognition, but the features do not establish which bands will be visible.
Isoform 2 lacks UniProt residues 131–151, a 21-residue segment. Check whether the antibody recognizes that region when comparing bands; a shorter sequence alone does not establish its apparent molecular weight.
Isoform 3 replaces UniProt residues 98–129 with another 32-residue sequence and lacks residues 130–184. Check the antibody epitope against both changes: an antibody targeting the deleted region would not recognize isoform 3.
Isoform 2 lacks UniProt residues 131–151, a 21-residue segment. Check whether the antibody recognizes that region when comparing bands; a shorter sequence alone does not establish its apparent molecular weight.
Isoform 3 replaces UniProt residues 98–129 with another 32-residue sequence and lacks residues 130–184. Check the antibody epitope against both changes: an antibody targeting the deleted region would not recognize isoform 3.
Do annotated modifications explain a shifted BMF band?
PTM · No modified residues or glycosylation sites are listed in the supplied features. The features therefore do not support assigning a shifted band to a specific modification; they also do not rule out unlisted modifications.
Does this guide establish induction of BMF?
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 BMF?
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 A02432-1 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 can I quantify BMF when multiple bands appear?
Quantitation · Decide whether the antibody epitope is shared by the listed isoforms before combining band signals. For isoform-specific comparisons, quantify resolved bands separately and report which band was measured. The supplied features do not establish the identity of any observed band.
How should unexpected BMF bands be interpreted?
Interpretation · First compare their pattern with the three listed isoforms and the antibody epitope. A band at another position cannot be assigned to an isoform or modification from these features alone; no observed band size is supplied.
Boster reagents
BMF 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-Bcl-2-modifying factor Bmf Antibody
Cat # A02432-1
Real WB data
Anti-Bmf Rabbit Monoclonal Antibody
Cat # M02432-1
Real WB data
Anti-Bcl-2-modifying factor Bmf Antibody
Cat # A02432
Three the supplier anti-BMF antibodies have Western blot images using 293, Ramos, or HepG2 cell lysates. All list human reactivity; two list mouse and one lists rat. The supplied captions show specific lysate examples, not WB validation across every listed species.
Which to pick: Choose A02432-1 for its 293 lysate blot at 1 and 2 μg/mL, A02432 for HepG2 at 2.5 and 5 μg/mL, or M02432-1 for the Ramos blot and listed rat reactivity. Match the listed reactivity and demonstrated sample to your experiment.
Source: BosterBio BMF gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96LC9.
- Human Protein Atlas. BMF tissue expression.
- PMC9937703 — target-verified WB comparison
- PMC6481009 — target-verified WB comparison
- PMC3639396 — target-verified WB comparison