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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
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- Product number
- 44-1350G - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-Btk (Tyr223) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of lysates prepared from EB1 cells using rabbit anti-BTK (pY223) antibody (Product # 44-1350G). Cells left untreated (lane 1) or treated with hydrogen peroxide (lanes 2-5) were resolved on a 10 percent polyacrylamide gel and transferred to PVDF. The membrane was treated with either no peptide (lane 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (lane 3), a generic phosphotyrosine containing peptide (lane 4), or the phosphopeptide immunogen (lane 5). Prior to incubation with the primary antibody, lane 6 was treated with Lambda phosphatase.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Phospho-Btk (Tyr223) Polyclonal Antibody (Product # 44-1350G) and a 75 kDa band corresponding to Phospho-Btk along with an uncharacterized band (*) at ~160 kDa were observed in the treated panel. Whole cell extracts (40 µg lysate) of Ramos (Lane 1), Ramos treated with hydrogen peroxide (100mM, 20 minutes) (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
