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SREBF1 / Sterol regulatory element-binding protein 1 · Western blot design guide
Design a Western Blot for SREBF1
Real validated SREBF1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SREBF1 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
SREBF1 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 | ~121.7 kDa | |
| Observed band | Approximately 122 kDa | |
| Gel | 10% (catalog A00282-2) | |
| 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 | Phosphorylated + Ubl conjugation | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 6 isoform(s) |
Section 1
Real Curated SREBF1 Western Blot Protocols
The A00282-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human Jurkat, rat RH-35 (catalog A00282-2) |
| Gel % | 10% (catalog A00282-2) |
| Load | 30 ug; reducing conditions (catalog A00282-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00282-2) |
| Membrane | nitrocellulose membrane (catalog A00282-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00282-2) |
| Primary antibody | A00282-2 · 1:1000 (catalog A00282-2) |
| Primary incubation | overnight at 4°C (catalog A00282-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00282-2) |
| Secondary incubation | 1.5 hour at RT (catalog A00282-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00282-2) |
| Detection | ECL (catalog A00282-2) |
Section 2
What Is the Expected SREBF1 Western Blot Band Size?
SREBF1 has a predicted mass of 121.7 kDa and an observed band at approximately 122 kDa; the evidence does not establish a cause for their small difference.
| Band at approximately 122 kDa | matches the observed SREBF1 band and its 121.7 kDa predicted mass |
| Additional bands at different positions | could reflect SREBF1 splice isoforms; their migration is unreported |
| Closely spaced bands | could reflect differences in phosphorylation; a visible shift is unproven |
| Weak or absent band in a soluble fraction | could reflect SREBF1 association with ER and Golgi membranes |
💡Expected SREBF1 appearanceThe predicted full-length mass is 121.7 kDa, and an approximately 122 kDa band was observed in whole-cell lysates; confirm its identity with an appropriate antibody or SREBF1 depletion control.
| Predicted full-length mass | 121.7 kDa predicted; an approximately 122 kDa band was observed |
| SREBP-1A and SREBP-1B splice isoforms | may differ in size; individual masses and migration are unreported |
| SREBP-1C and isoform 4 | may differ in size; individual masses and migration are unreported |
| SREBP-1aDelta and SREBP-1cDelta splice isoforms | may differ in size; individual masses and migration are unreported |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | membrane-associated SREBF1 may be poorly recovered | check membrane extraction and use an appropriate positive-control lysate |
| Band higher than expected | an isoform or altered migration may be involved; the cause is unestablished | compare with the approximately 122 kDa reference band and confirm by SREBF1 depletion |
| Band lower than expected | an isoform or sample fragmentation may be involved; the cause is unestablished | use fresh lysate and confirm the band by SREBF1 depletion |
| Multiple bands | splice isoforms or phosphorylation may contribute, but distinct bands are unproven | check which bands respond to SREBF1 depletion |
| Weak or no signal | incomplete recovery of membrane-associated SREBF1 may reduce signal | check extraction and compare with the whole-cell lysates used to observe the 122 kDa band |
Experiment evidence
Western blot images showing SREBF1 band
A00282-2 · SREBF1
A00282 · SREBF1
A00282-1 · SREBF1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SREBF1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SREBF1 in Western blot, you can use adrenal gland tissue, which shows high expression in HPA.
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 such as Ponceau alongside the samples.
⚠️Feasibility: SREBF1 is a membrane protein, so effective membrane protein extraction may be needed for a strong signal.
HPA tissue expression evidence for SREBF1
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 → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | Medium | Protein (IHC) | HPA → |
| Lung | macrophages | Medium | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | Medium | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SREBF1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SREBF1, answered from its protein features.
How should SREBF1 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.
Which SREBF1 isoforms could affect band size?
Isoforms · UniProt lists six isoforms. SREBP-1C has a shorter N-terminal alternative sequence; SREBP-1B and SREBP-1C replace residues 1035–1147; and both Delta isoforms lack residues 471–1147. Compare the isoform expressed in your sample before assigning a band by size.
Check the antibody epitope against the isoform you intend to measure. An epitope within UniProt residues 471–1147 is absent from both Delta isoforms; the N terminus and residues 1035–1147 also vary among listed isoforms. Match any reported antibody numbering to the UniProt sequence before comparing coordinates.
Check the antibody epitope against the isoform you intend to measure. An epitope within UniProt residues 471–1147 is absent from both Delta isoforms; the N terminus and residues 1035–1147 also vary among listed isoforms. Match any reported antibody numbering to the UniProt sequence before comparing coordinates.
Which phosphorylation sites matter when interpreting SREBF1 bands?
PTM · UniProt lists phosphoserines at residues 98, 117, 337, 338, 396, 402, 457, and 1060. It attributes 337, 338, and 402 to SIK1, and 396 to AMPK. These are UniProt coordinates, which may differ from antibody or paper numbering. Their presence does not establish a visible band shift.
Does this guide establish induction of SREBF1?
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 SREBF1 Western blot?
Transfer · For the approximately 122 kDa, multi-pass membrane protein, consider a wet transfer setup suitable for high-mass proteins. Verify transfer around 122 kDa before interpreting a weak signal; the supplied features do not specify transfer conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00282-2 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 SREBF1 bands be quantified?
Quantitation · Quantify the approximately 122 kDa band consistently across samples, using the same band boundary and a documented normalization method. If additional bands appear, measure them separately until their identity is established. The six isoforms and membrane localization make an antibody's recognized region relevant to interpretation.
Should SREBF1 run above its predicted mass?
Interpretation · The predicted mass is 121.7 kDa, and the supplied observed band is approximately 122 kDa. These values agree at this precision. The listed modifications and isoforms alone do not establish a visible shift.
First check whether the antibody recognizes the relevant isoform sequences: the Delta isoforms lack UniProt residues 471–1147, and other isoforms have terminal changes. SREBF1 is also listed in ER and Golgi membranes. Band position alone cannot identify an isoform, a phosphorylation state, or a processing product.
First check whether the antibody recognizes the relevant isoform sequences: the Delta isoforms lack UniProt residues 471–1147, and other isoforms have terminal changes. SREBF1 is also listed in ER and Golgi membranes. Band position alone cannot identify an isoform, a phosphorylation state, or a processing product.
Boster reagents
SREBF1 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-SREBF1 Antibody
Cat # A00282-2
Real WB data
Anti-SREBF1 Antibody
Cat # A00282
Real WB data
Anti-SREBP-1 SREBF1 Antibody
Cat # A00282-1
Three the supplier anti-SREBF1 antibodies have Western blot images. A00282-2 shows an approximately 122 kDa band in human HepG2 and Jurkat and rat RH-35 lysates; A00282 and A00282-1 have images in Daudi and TNF-treated JK lysates, respectively. Evidence is limited to these reported contexts.
Which to pick: For human or rat samples, A00282-2 has the clearest reported WB sample and band details. A00282 lists human and mouse reactivity, but its WB caption describes only Daudi lysate. A00282-1 lists human and rat reactivity and shows TNF-treated JK lysate with a peptide-treated lane.
Source: BosterBio SREBF1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P36956.
- Human Protein Atlas. SREBF1 tissue expression.
- PMC10256812 — target-verified WB comparison
- PMC9178675 — target-verified WB comparison
- PMC11801665 — target-verified WB comparison