Anti-Peroxiredoxin 1/PAG antibody [EPR5434] (ab109506)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5434] to Peroxiredoxin 1/PAG
- Suitable for: WB, ICC/IF, Flow Cyt
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-Peroxiredoxin 1/PAG antibody [EPR5434]
See all Peroxiredoxin 1/PAG primary antibodies -
Description
Rabbit monoclonal [EPR5434] to Peroxiredoxin 1/PAG -
Host species
Rabbit -
Specificity
Corresponding to residues in Human Peroxiredoxin 1/PAG -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanWB Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat, 293T, K562 or U87-MG cell lysate. IHC-P: Human liver or kidney tissue. IF/ICC: HeLa cells.
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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. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR5434 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-Peroxiredoxin 1/PAG antibody [EPR5434] (ab109506) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : PRDX1 knockout HEK293T cell lysate
Lane 3 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 22 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab109506 observed at 26 kDa. Red - loading control ab8245 observed at 36 kDa.
ab109506 Anti-Peroxiredoxin 1/PAG antibody [EPR5434] was shown to specifically react with Peroxiredoxin 1/PAG in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266842 (knockout cell lysate ab257040) was used. Wild-type and Peroxiredoxin 1/PAG knockout samples were subjected to SDS-PAGE. ab109506 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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® 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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ICC/IF image of ab109506 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab109506, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Peroxiredoxin 1/PAG with unpurified ab109506 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Peroxiredoxin 1/PAG knockout HAP1 cell lysate (20 µg)
Lane 3: A431 cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109506 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.ab109506 was shown to specifically react with Peroxiredoxin 1/PAG when Peroxiredoxin 1/PAG knockout samples were used. Wild-type and Peroxiredoxin 1/PAG knockout samples were subjected to SDS-PAGE. ab109506 and ab8245 (loading control to GAPDH) were both diluted 1/10 000 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 1/10 000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-Peroxiredoxin 1/PAG antibody [EPR5434] (ab109506) at 1/10000 dilution
Lane 1 : 293T cell lysate
Lane 2 : K562 cell lysate
Lane 3 : U87-MG cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 22 kDa
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