Human PACT Antibody

R&D Systems, part of Bio-Techne | Catalog # AF1980

R&D Systems, part of Bio-Techne
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AF1980
AF1980-SP
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Key Product Details

Species Reactivity

Human

Applications

Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all PACT Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human PACT
Ser2-Lys313
Accession # O75569

Specificity

Detects human PACT in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human PACT Antibody

Detection of Human PACT antibody by Western Blot.

Detection of Human PACT by Western Blot.

Western blot shows lysates of K562 human chronic myelogenous leukemia cell line and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human PACT Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1980) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for PACT at approximately 32 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human PACT antibody by Simple WesternTM.

Detection of Human PACT by Simple WesternTM.

Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for PACT at approximately 42 kDa (as indicated) using 10 µg/mL of Goat Anti-Human PACT Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1980) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for Human PACT Antibody

Application
Recommended Usage

Simple Western

10 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line

Western Blot

1 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line and HeLa human cervical epithelial carcinoma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: PACT

PACT (PKR Activator; also HSD14, PRKRA and RAX [in mouse]) is a 34-37 kDa member of the PRKRA family of molecules. It is a ubiquitously expressed, cytoplasmic, 313 amino acid (aa) protein that contains three discrete double-stranded (ds) RNA binding motifs (aa 31-101; 126-194; 240-308). It exists as a monomer, homodimer, and heterodimer with at least two partners, TRBP and Dicer. Two isoform variants are known to exist, including one that utilizes an alternative start site at Met26, and another that shows a 10 aa substitution for aa 1-21. PACT is perhaps best known for its relationship with PKR, a 52-58 kDa Ser/Thr kinase that downmodulates protein synthesis and initiates apoptosis. PKR is constitutively silent, and activated via two principal ways; the first is through binding to (viral) dsRNA, and the second is through binding to an activator, PACT. PACT is kept in check upstream through dimerization with at 48-49 kDa regulatory protein termed TRBP. Following growth factor withdrawl or exposure to bacterial products such as LPS, PACT is phosphorylated at Ser287, and this inactive PACT:TRBP heterodimer dissociates, freeing up PACT to bind to PKR. PACT's interaction with PKR induces an autophosphorylation event on PKR. This allows for an activated PKR interaction with eIF2a, causing its phosphorylation and a subsequent reduction of protein synthesis. PACT may also act independently of PKR, and serve as a component of RISC, a complex that downregulates mRNA translation. Within this complex, PACT likely associates with Dicer in concert with Argonaute. Full-length PACT shares 98% aa sequence identity with the mouse ortholog to human PACT termed RAX.

Alternate Names

DYT16, HSD14, PKR-Associating Protein X, PRKRA, RAX

Entrez Gene IDs

8575 (Human); 23992 (Mouse); 311130 (Rat)

Gene Symbol

PRKRA

UniProt

O75569

Additional PACT Products

Product Documents for Human PACT Antibody

Product Datasheets

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human PACT Antibody

For research use only

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