Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
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Validation data
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- Product number
- AF3227 - Provider product page
- Provider
- R&D Systems
- Product name
- Human/Mouse 4EBP1 Antibody
- Antibody type
- Polyclonal
- Description
- Antigen Affinity-purified. Detects human and mouse 4EBP1.
- Reactivity
- Human, Mouse
- Host
- Goat
- Conjugate
- Unconjugated
- Antigen sequence
Q13541
- Isotype
- IgG
- Vial size
- 100 ug
- Concentration
- LYOPH
- 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.
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Supportive validation
- Submitted by
- R&D Systems (provider)
- Main image
- Experimental details
- 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.
- Submitted by
- R&D Systems (provider)
- Main image
- Experimental details
- 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.
- Submitted by
- R&D Systems (provider)
- Main image
- Experimental details
- 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.