Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-37824 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-ERK1/ERK2 (Thr202) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- A suggested positive control for Western blot is 293 cells; suggested positive control for IHC is human breast carcinoma; suggested positive control for ICC/IF is Hela cells.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of extracts from 293 cells, untreated or treated with PMA for the indicated times, using p44/42 MAP Kinase (pThr202) polyclonal antibody (Product # PA5-37824).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- WB was performed on whole cell extracts (30 µg) of A-431 (1), A-431 treated with EGF (200 ng/mL for 10 minutes) (2), A-431 treated with Afatinib followed by EGF (0.5uM of Afatinib for 6hrs, 200 ng/mL for 10 minutes) (3), MCF7 (4) and MCF7 treated with PMA (200 ng/mL for 10 minutes) (5), HeLa (6) and HeLa treated with PMA (200 ng/mL for 10 minutes) (7), NIH/3T3 (8) and NIH/3T3 treated with PMA (200 ng/mL for 10 minutes) (9). The blot was probed with Anti-Phospho-ERK1/ERK2 (Thr202) Rabbit Polyclonal Antibody (Product # PA5-37824, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). 44, 42 kDa band corresponding to Phospho-ERK1/ERK2 (Thr202) was detected and increased upon EGF and PMA treatment across cell lines tested and pre-treatment with Afatinib (antagonist) resulted in inhibition of Phospho-ERK1/ERK2 (Thr202) in A-431 cell line upon EGF treatment. Samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ System (Product # EI0002) and Novex® Sharp Standard (Product # LC5800). Proteins were wet transferred onto a nitrocellulose membrane. Primary and secondary Antibodies were used following blocking with 5 % skimmed milk. Detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of Phospho-ERK1/ERK2 (Thr202) in methanol-fixed Hela cells cytoplasmic, nuclear staining, using Phospho-ERK1/ERK2 (Thr202) Polyclonal Antibody (Product # PA5-37824).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Phospho-ERK1/ERK2 (Thr202) in paraffin-embedded Human breast carcinoma tissue using Phospho-ERK1/ERK2 (Thr202) Polyclonal Antibody (Product # PA5-37824) (left) or the same antibody preincubated with blocking peptide (right).