Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5513] to PI 3 Kinase p85 alpha
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-PI 3 Kinase p85 alpha antibody [EPR5513]
See all PI 3 Kinase p85 alpha primary antibodies -
Description
Rabbit monoclonal [EPR5513] to PI 3 Kinase p85 alpha -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species WB Human -
Immunogen
Synthetic peptide corresponding to a region within the N terminus of Human PI 3 Kinase p85 alpha (UniProt ID: P27986)
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Positive control
- HeLa, Jurkat, Raji, K562, MCF7, C6, Raw 264.7, PC12, NIH3T3 cell lysates
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 5.50 x 10 -12 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR5513 -
Isotype
IgG -
Research areas
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PI 3 Kinase p85 alpha knockout HAP1 cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: THP1 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab133595 was shown to specifically react with PI 3 Kinase p85 alpha when PI 3 Kinase p85 alpha knockout samples were used. Wild-type and PI 3 Kinase p85 alpha knockout samples were subjected to SDS-PAGE. ab133595 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PIK3R1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab133595 was shown to react with PI 3 Kinase p85 alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265116 (knockout cell lysate ab257029) was used. Wild-type HeLa and PIK3R1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133595 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : Raji cell lysate
Lane 3 : K562 cell lysate
Lane 4 : MCF7 cell lysate
Lane 5 : C6 cell lysate
Lane 6 : Raw 264.7 cell lysate
Lane 7 : PC12 cell lysate
Lane 8 : NIH3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled Goat anti Rabbit at 1/2000 dilution
Predicted band size: 83 kDa
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