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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
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- Product number
- 711351 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-HSP27 (Ser78) Recombinant Polyclonal Antibody (4HCLC)
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 4HCLC
- 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
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis was performed on Whole cell extracts (30 µg lysate) of HeLa, HeLa (UV for 40 min). The blots were probed with Anti-Hsp27 (pS78) Recombinant Rabbit Polyclonal Antibody (Product # 711351, 1-2 µg/mL) (Lane 1, 2). To confirm the specificity of Anti-Anti-Hsp27 (pS78), competition was performed with the phospopeptide (10 µg/mL) (Lane 3, 4) and non phospopeptide (10 µg/mL) (Lane 5, 6). A 27 kDa band corresponding to Anti-Hsp27 (pS78) was observed in HeLa (with increase in expression upon UV treatment) when probed with the antibody alone or with non-phospho peptide competition. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), 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).
