Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3224] to PACT (PKR activating protein) / PRKRA
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224]
See all PACT (PKR activating protein) / PRKRA primary antibodies -
Description
Rabbit monoclonal [EPR3224] to PACT (PKR activating protein) / PRKRA -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human -
Immunogen
Synthetic peptide within Human PACT (PKR activating protein)/ PRKRA aa 300-400 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa, K562, Jurkat and HepG2 whole cell lysate (ab7900). IHC-P: Human brain tissue.
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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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
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
EPR3224 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PRKRA knockout HeLa cell lysate
Lane 3 : K-562 cell lysate
Lane 4 : HepG2 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: 34 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab75749 observed at 36 kDa. Red - loading control ab8245 observed at 36 kDa.
ab75749 Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] was shown to specifically react with PACT in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266806 (knockout cell lysate ab258141) was used. Wild-type and PACT knockout samples were subjected to SDS-PAGE. ab75749 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)
ab75749 at 1/100 dilution staining PACT (PKR activating protein) / PRKRA in paraffin-embedded human brain tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PACT (PKR activating protein)/PRKRA knockout HAP1 cell lysate (20 µg)
Lane 3: K562 cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab75749 observed at 36 kDa. Red - loading control, ab18058, observed at 124 kDa.ab75749 was shown to specifically react with PACT (PKR activating protein)/PRKRA when PACT (PKR activating protein)/PRKRA knockout samples were used. Wild-type and PACT (PKR activating protein)/PRKRA knockout samples were subjected to SDS-PAGE. ab75749 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749) at 1/1000 dilution
Lane 1 : K562 cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : HepG2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDa
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