western blot antibody customer review for p63 polyclonal antibody (stj94912)
TRANSCRIPT
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HOW DID OUR ANTIBODY PERFORM?
ANTIBODY CUSTOMER REVIEW: p63 Polyclonal Antibody (STJ94912)
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What was used?Primary antibody: STJ94912 p63 antibody
Provider: St John's Laboratory Ltd.
Dilution ratio: 1:500
Application: Western Blot
Materials for validation: Human corneal limbal stem cells
Click for product data sheet PDF
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What Was The Protocol?Treatment of Materials When cells were cultured to 70%-80% plating density, lysis buffer with protease inhibitors
(Roche) and equal volume of 2X loading buffer were added for 10 minutes in a water bath at 100˚C.
Gel Electrophoresis 10% polyacrylamide gel, voltage 80V for 15 minutes, then 140 V for 45 minutes (total 60 minutes).
Transfer 0.45 µm NC membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 minutes.
Blocking Odyssey Blocking Buffer (PBS) for 1 hour.
Primary Antibody Probing Primary antibody was added to the solution of Odyssey Blocking Buffer (PBS) plus 0.01% Tween 20, 4˚C overnight.
Membrane Wash Odyssey Blocking Buffer (PBS) plus 0.01% Tween-20 for 4 times.
Secondary Antibody Probing Secondary antibody was added to the solution of Odyssey Blocking Buffer (PBS) plus 0.01% Tween-20 and 0.02% SDS with 1:10000 dilution ratio, for 1 hour.
Membrane Wash Odyssey Blocking Buffer (PBS) plus 0.01% Tween-20 for 4 times.
Visualization Odyssey CLx Near-Infrared Imaging System.
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What was the protocol?10% polyacrylamide gel, voltage 80V for 15 minutes, then 140 V for 45 minutes (total 60 minutes).
Transfer information: 0.45 µm NC membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 minutes.
No. Antigen Loading amount
Primary antibody
Primary antibody
dilution ratio
Secondaryantibody
dilution ratioTarget band KD Visualization
time
1P0 human
corneal limbal stem cells (P0)
20µg (total protein)
STJ94912 p63 antibody
1:500
1:0000 ~77KD 60s
2P1 human
corneal limbal stem cells (P1)
20µg (total protein) ~77KD 60s1:0000
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What were the results?
Figure: Lane1: P0; Lane 2: P1
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What did the customer think?
Antibody Specificity:
Antibody RPPA Rating:
Testing results were provided independently by F. Abidin at Newcastle University. They stated “In human corneal limbal stem cells, there were target bands detected by STJ94912 p63 antibody”
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JOIN THE ANTIBODY VALIDATION PROJECT
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1. Order any STJ9 catalogued product using this online form.
2. Test it in your laboratory using your preferred method.
3. Return a fully completed review form to receive rewards.
…in 3 simple steps…
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@StJohnsLabs
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