Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-34808 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- RIP2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: HeLa, Molt-4. Predicted reactivity: Rat (80%), Bovine (82%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.87 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 was performed using Anti-RIP2 Polyclonal Antibody (Product # PA5-34808) and a 60 kDa band corresponding to Receptor-interacting serine/threonine-protein kinase 2 was observed across the cell lines tested except MOLT-4, Jurkat and MCF 10A which are reported to be negative. Whole cell extracts (30 µg lysate) of Raji (Lane 1), HeLa (Lane 2), U-87 MG (Lane 3), A-431 (Lane 4), THP-1 (Lane 5), MOLT-4 (Lane 6), Jurkat (Lane 7), MCF7 (Lane 8) and MCF 10A (Lane 9) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using RIP2 Polyclonal Antibody (Product # PA5-34808). Sample (30 µg of whole cell lysate). Lane A: Molt-4 . 7.5% SDS PAGE. RIP2 Polyclonal Antibody (Product # PA5-34808) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-RIP2 Polyclonal Antibody (Product # PA5-34808) and a 60 kDa band corresponding to Receptor-interacting serine/threonine-protein kinase 2 was observed across the cell lines tested except MOLT-4, Jurkat and MCF 10A which are reported to be negative. Whole cell extracts (30 µg lysate) of Raji (Lane 1), HeLa (Lane 2), U-87 MG (Lane 3), A-431 (Lane 4), THP-1 (Lane 5), MOLT-4 (Lane 6), Jurkat (Lane 7), MCF7 (Lane 8) and MCF 10A (Lane 9) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Receptor-interacting serine/threonine-protein kinase 2 was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with RIP2 Polyclonal Antibody (Product # PA5-34808) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic and Nuclear localization. Panel e represents MCF 10A cells with decreased expression of RIP2. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded A549 xenograft, using RIP2 (Product # PA5-34808) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.