This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
B4GALT1 / Beta-1,4-galactosyltransferase 1 · Western blot design guide
Design a Western Blot for B4GALT1
Source-linked B4GALT1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-B4GALT1 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
B4GALT1 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~43.9 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 | Glycosylated | |
| Caveat | Knockout control | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked B4GALT1 Western Blot Protocol Options
The A03993-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts from normal and B4GALT1 knockout HeLa cells, (catalog A03993-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A03993-1) |
| Primary antibody | A03993-1 · 1:3000 (catalog A03993-1) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A03993-1) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A03993-1) |
Section 2
What Is the Expected B4GALT1 Western Blot Band Size?
B4GALT1 has a predicted mass of 43.9 kDa; glycosylation, homodimerization, and isoforms could alter bands, but their migration effects are not demonstrated here.
| Band near 43.9 kDa | Consistent with the predicted protein mass; confirm identity with the knockout control. |
| Band above 43.9 kDa | N-linked glycosylation at Asn113 could affect migration, but no shift is established. |
| Band near twice the monomer size under nonreducing conditions | Could represent the annotated homodimer. |
| Several bands at different positions | Long and Short isoforms are annotated, but distinct migration is unverified. |
| Little or no band in a soluble lysate fraction | B4GALT1 is a Golgi membrane protein and may be poorly recovered in that fraction. |
💡Expected B4GALT1 appearanceUniProt predicts 43.9 kDa, but no empirical band size is supplied; glycosylation, homodimerization, and isoforms may affect the pattern, so use the B4GALT1 knockout control to identify bands.
| UniProt predicted mass | Sets a 43.9 kDa reference for the monomer. |
| N-linked glycosylation at Asn113 | Could change apparent migration; the size of any shift is unknown. |
| Homodimer formation | Could produce a band near twice the monomer size under nonreducing or incompletely reducing conditions. |
| Long and Short isoforms | Could differ in apparent size, but their masses and band separation are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Golgi membrane protein may be poorly extracted. | Check membrane recovery and compare with the B4GALT1 knockout control. |
| Band higher than expected | Homodimer may persist if reduction is incomplete; Asn113 glycosylation may also affect migration. | Compare reducing conditions and confirm band identity with knockout lysate. |
| Band lower than expected | Could reflect an isoform, but its migration is unknown. | Compare with knockout lysate before assigning the band. |
| Multiple bands | Long and Short isoforms or differing Asn113 glycosylation are possible; neither pattern is established. | Compare each band with knockout lysate and assess glycosylation if needed. |
| Weak or no signal | Poor recovery of this Golgi membrane protein is possible. | Check extraction and loading, then compare with knockout lysate. |
Experiment evidence
Western blot images showing B4GALT1 band
A03993-1 · B4GALT1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for B4GALT1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for B4GALT1 in Western blot, you can use adrenal gland tissue, which shows high HPA 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 such as Ponceau alongside the samples.
⚠️Feasibility: Adipose tissue offers a reported negative control, though B4GALT1 secretion may affect the signal retained in tissue lysate.
HPA tissue expression evidence for B4GALT1
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 → |
| Bronchus | basal cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | 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 → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced B4GALT1 Western Blot Tips
Deeper troubleshooting and optimisation questions for B4GALT1, answered from its protein features.
How should B4GALT1 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 the Long and Short isoforms affect band interpretation?
Isoforms · The Short isoform lacks residues 1–13 of the Long isoform. This sequence difference could produce distinct bands, but their separation is not established. Check whether the antibody recognizes both isoforms before assigning bands.
How can glycosylation be tested when interpreting B4GALT1 bands?
PTM · Compare matched samples before and after N-glycan removal. The annotated site is Asn113 in the supplied UniProt sequence; coordinates may differ in isoform or antibody numbering. A change in migration would support a glycosylation contribution, but is not guaranteed.
Does this guide establish induction of B4GALT1?
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 B4GALT1 Western blot?
Transfer · B4GALT1 is a predicted 43.9 kDa single-pass membrane protein. Choose a transfer setup validated around 44 kDa and check transfer with total-protein staining. The supplied features do not establish whether wet or semidry transfer performs better.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03993-1 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 B4GALT1 bands be quantified?
Quantitation · Quantify bands consistently across samples and normalize to a suitable loading measure. Because Long and Short isoforms are annotated, decide whether the measurement represents one resolved band or their combined signal, and use an antibody that recognizes the intended forms.
Why might B4GALT1 migrate differently from its predicted 43.9 kDa mass?
Interpretation · B4GALT1 has an annotated N-linked glycosylation site at Asn113. Glycosylation could affect migration, but this feature alone does not establish a visible shift. No empirical band size is supplied, so verify any difference experimentally.
B4GALT1 is annotated as a homodimer and can associate with LALBA to form lactose synthase. Those features do not prove a higher Western-blot band is a complex. Compare reducing and nonreducing preparations and confirm band identity with appropriate controls.
B4GALT1 is annotated as a homodimer and can associate with LALBA to form lactose synthase. Those features do not prove a higher Western-blot band is a complex. Compare reducing and nonreducing preparations and confirm band identity with appropriate controls.
Boster reagents
B4GALT1 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-B4GALT1 Antibody
Cat # A03993-1
A03993-1 is listed as reactive with human samples. Its Western blot image compares normal and B4GALT1-knockout HeLa cell extracts using a 1:3000 antibody dilution. The supplied validation evidence is limited to this reported context.
Which to pick: A03993-1 is the only listed B4GALT1 antibody. It has a Western blot image from normal and knockout HeLa extracts, making it the documented option for that sample context.
Source: BosterBio B4GALT1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P15291.
- Human Protein Atlas. B4GALT1 tissue expression.
- PMC10618527 — target-verified WB comparison
- PMC4982324 — target-verified WB comparison
- PMC10259029 — target-verified WB comparison