Anti-PP-T antibody [12F7] (ab223367)
![Anti-PP-T antibody [12F7] (ab223367)](https://www.abcam.com/ps/products/223/ab223367/Images/ab223367-410178-anti-pp-t-antibody-12f7-western-blot-human-knockout-lysate.jpg "Anti-PP-T antibody [12F7] (ab223367)")
Key features and details
- Mouse monoclonal [12F7] to PP-T
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-PP-T antibody [12F7]
See all PP-T primary antibodies -
Description
Mouse monoclonal [12F7] to PP-T -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Recombinant full length protein corresponding to Human PP-T aa 1 to the C-terminus.
Database link: P53041 -
Positive control
- WB: A431, HEK-293 and Jurkat whole cell lysate (ab7899). ICC/IF: HeLa and HEK-293 cells.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.09% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), PBS -
Concentration information loading... -
Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
12F7 -
Isotype
IgG1 -
Research areas
Images
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Western blot - Anti-PP-T antibody [12F7] (ab223367)All lanes : Anti-PP-T antibody [12F7] (ab223367) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : PPP5C knockout HEK293T cell lysate
Lane 3 : A431 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Predicted band size: 56 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab223367 observed at 58 kDa. Red - loading control ab181602 observed at 36 kDa.
ab223367 Anti-PP-T antibody [12F7] was shown to specifically react with PP-T in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266507 (knockout cell lysate ab258136) was used. Wild-type and PP-T knockout samples were subjected to SDS-PAGE. ab223367 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)](https://www.abcam.com/ps/products/223/ab223367/Images/ab223367-283068-anti-pp-t-antibody-12f7-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)4% formaldehyde-fixed HeLa (human epithelial cell line from cervix adnocarcinoma) cells stained for PP-T (green) using ab223367 at 1/100 dilution for 60 minutes at room temperature in ICC/IF, followed by Goat Anti-Mouse ATTO 488 at 1/100 dilution for 60 minutes at room temperature. Counterstain: DAPI (blue) nuclear stain at 1/5000 for 5 minutes room temperature.
(A) DAPI nuclear stain (B) ab223367 (C) Merge.
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![Western blot - Anti-PP-T antibody [12F7] (ab223367)](https://www.abcam.com/ps/products/223/ab223367/Images/ab223367-283067-anti-pp-t-antibody-12f7-western-blot.png)
Western blot - Anti-PP-T antibody [12F7] (ab223367)All lanes : Anti-PP-T antibody [12F7] (ab223367) at 1/500 dilution
Lane 1 : A431 (human epidermoid carcinoma cell line) cell lysate
Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate
Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG HRP at 1/200 dilution
Developed using the ECL technique.
Predicted band size: 56 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?
Exposure time: 6 minutesBlocking buffer: 5% skim milk.
Primary and secondary antibodies were incubated for 1 hour at room temperature.
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![Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)](https://www.abcam.com/ps/products/223/ab223367/Images/ab223367-283066-anti-pp-t-antibody-12f7-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (ab223367)2% formaldehyde-fixed HEK-293 (human epithelial cell line from embryonic kidney) cells stained for PP-T (green) using ab223367 at 1/50 dilution in ICC/IF, followed by Goat Anti-Mouse Alexa Fluor 488 at 1/100 dilution for 60 minutes at RT. Counterstain: DAPI (blue) nuclear stain.
(A) DAPI nuclear stain (B) ab223367 (C) Merge.
