Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
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Validation data
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- Product number
- 720115 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-Cyclin E (Ser88) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), HEK-293 treated with hydroxyurea (2mM Hydroxy urea for 24hrs) (Lane 2), HEL (Lane 3), HEL treated with hydroxyurea (2mM Hydroxy urea for 24hrs) (Lane 4), HeLa (Lane 5), HeLa treated with hydroxyurea (2mM Hydroxy urea for 24hrs) (Lane 6). The blots were probed with Anti-Cyclin E (pS88) Rabbit Polyclonal Antibody (Product # 720115, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 35 kDa band corresponding to Cyclin E (pS88) which possibly represents the lower molecular weight isoform of Cyclin E upon phosphorylation at S88 is observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12% Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis HeLa cells were fixed and permeabilized for detection of Cyclin E (pS88) using Cyclin E (pS88) Rabbit Polyclonal antibody (Product # 720115, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Detection and localization Cyclin E (pS88) (green) in the cytoplasm can be clearly observed in cells treated with serum starvation (SS) as compared to untreated cells. Antibody specificity was demonstrated by competition with the Cyclin E (pS88) peptide, which results in total loss of signal. No competition was observed with the non-phospho peptide.