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Human/Mouse 4EBP1 Antibody
Catalog # AF3227 | R&D Systems, Inc. a Bio-Techne Brand
Key Product Details
Validated by
Knockout/Knockdown
Species Reactivity
Human, Mouse
Applications
Knockout Validated, Simple Western, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
Product Summary for Human/Mouse 4EBP1 Antibody
Immunogen
E. coli-derived recombinant human 4EBP1
Ser2-Ile118
Accession # Q13541
Ser2-Ile118
Accession # Q13541
Specificity
Detects human and mouse 4EBP1.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Human/Mouse 4EBP1 Antibody
Detection of Human/Mouse 4EBP1 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and C2C12 mouse myoblast cell line. PVDF membrane was probed with 0.5 µg/mL of Human/Mouse 4EBP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3227) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for 4EBP1 at approximately 18-20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human 4EBP1 by Simple WesternTM.
Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for 4EBP1 at approximately 26 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse 4EBP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3227) 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.
Western Blot Shows Human 4EBP1 Specificity by Using Knockout Cell Line.
Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and 4EBP1 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse 4EBP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3227) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for 4EBP1 at approximately 17-20 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1. New adjunct appears with knockout cell line.Applications for Human/Mouse 4EBP1 Antibody
Application
Recommended Usage
Knockout Validated
4EBP1
is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in
4EBP1 knockout HeLa cell line.
Simple Western
5 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line
Sample: HeLa human cervical epithelial carcinoma cell line
Western Blot
0.5 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and C2C12 mouse myoblast cell line
Sample: HeLa human cervical epithelial carcinoma cell line and C2C12 mouse myoblast 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.
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
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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: 4EBP1
The eukaryotic initiation factor 4E (eIF-4E) binding protein 1 (4EBP1) binds to eIF-4E to prevent translation initiation. Upon phosphorylation by the target of rapamycin (TOR) kinase, 4EBP1 releases eIF-4E and translation initiation ensues. 4EBP1 is also known as PHAS-1 (phosphorylated heat and acid stable protein regulated by insulin 1).
Product Documents for Human/Mouse 4EBP1 Antibody
Product Specific Notices for Human/Mouse 4EBP1 Antibody
For research use only
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