Customer Review For AXIN2 Polyclonal Antibody (STJ26209)

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

Transcript of Customer Review For AXIN2 Polyclonal Antibody (STJ26209)

Page 1: Customer Review For AXIN2 Polyclonal Antibody (STJ26209)

HOW DID OUR ANTIBODY PERFORM?

CUSTOMER REVIEW: AXIN2 Polyclonal Antibody (STJ26209)

Page 2: Customer Review For AXIN2 Polyclonal Antibody (STJ26209)

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.

Page 3: Customer Review For AXIN2 Polyclonal Antibody (STJ26209)

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.

Page 4: Customer Review For AXIN2 Polyclonal Antibody (STJ26209)

WHAT WAS THE PROTOCOL?

No. Antigen Loading amount

Primary antibody

Primary antibody

dilution ratio

Secondaryantibody

dilution ratio

1Human AXIN2

 10μg

STJ26209. AXIN2

antibody1:100 1:500

2 Non-specific 10μg Isotype rabbit IgG 1:100 1:500

Page 5: Customer Review For AXIN2 Polyclonal Antibody (STJ26209)

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.”

Page 6: Customer Review For AXIN2 Polyclonal Antibody (STJ26209)

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Page 7: Customer Review For AXIN2 Polyclonal Antibody (STJ26209)

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