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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [2]
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- Product number
- PA5-37737 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-SHC (Tyr349) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- A suggested positive control for Western blot is HepG2 cells; suggested positive control for ICC/IF is Hela cells.
- Reactivity
- Human, Mouse
- 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 HepG2 cells, untreated or treated with EGF, using Shc1 (pTyr349) polyclonal antibody (Product # PA5-37737).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Phospho-SHC (Tyr349) was performed using 90% confluent log phase A-431 cells treated with 200 ng/mL of EGF for 10 minutes. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-SHC (Tyr349) Rabbit Polyclonal Antibody (Product # PA5-37737) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree Celsius and then labelled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:100). Panel d represents the merged image showing membranous localization. Panel e represents cells treated with antagonist, Afatinib (1uM for 6hrs) followed by EGF (200 ng/mL for 10 minutes), showing no signal. Panel f shows untreated cells with no signal. Panel g represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of Phospho-SHC (Tyr349) in methanol-fixed Hela cells, using Phospho-SHC (Tyr349) Polyclonal Antibody (Product # PA5-37737).
