The Axin-related protein, Axin2, presumably plays an important role in the regulation of the stability of beta-catenin in the Wnt signaling pathway, like its rodent homologs, mouse conductin/rat axil. In mouse, conductin organizes a multiprotein complex of APC (adenomatous polyposis of the colon), beta-catenin, glycogen synthase kinase 3-beta, and conductin, which leads to the degradation of beta-catenin. To purchase this antibody, use the following link: http://www.stjohnslabs.com/axin2-antibody-p-67883?filter_name=STJ26209

1. HOW DID OUR
ANTIBODY PERFORM?
CUSTOMER REVIEW: AXIN2 Polyclonal Antibody (STJ26209)

2. WHAT WAS USED?
Primary antibody: AXIN2 Polyclonal Antibody
(STJ26209) 20μl.
Provider: St John's Laboratory
Dilution ratio: 1:100
Application: Flow Cytometry
Materials for validation: Human primary hepatocytes
Treatment of materials: Liver tissue was digested with
collagenase and cell were subjected to characterization
with FC.

3. WHAT WAS THE PROTOCOL?
Cell viability was detected by trypan blue.
Aliquots of 10^6 cell/ml were prepared in 0.5 % BSA in 1x PBBS
(labelling buffer).
Cell fixation: with 4 % paraformaldehyde at RT for 20˚C minutes.
Cell permealization: with -20°C cold 100% methanol at on ice for 30
minutes.
Primary antibody probing: primary antibody (or the isotype) was
added to the cell suspension 5μl/ 10^6 cells, at RT for 30 minutes in
dark.
Cells washed twice with labelling buffer 1 ml each and spun at 1000rpm
for 5 minutes.
Secondary antibody probing: secondary antibody to the cell
suspension 2μl/ 10^6 cells, at RT for 15 minutes in dark.
Cells washed twice with labelling buffer 1 ml each and spun at 1000rpm
for 5 minutes.
Visualization: FC visualization and analysis using Accuri C6.

4. WHAT WAS THE PROTOCOL?
No. Antigen
Loading
amount
Primary
antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
1
Human
AXIN2 10μg
STJ26209.
AXIN2
antibody
1:100 1:500
2 Non-specific 10μg
Isotype
rabbit IgG
1:100 1:500

5. WHAT DID THE CUSTOMER THINK?
Testing results were provided by Mogib Khedr from the
University of Southampton. They stated that “After
gating, hepatocytes in the area of interest were labelled
with Anti-Axin2 antibody and were distinct from unstained
or isotype control.”

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