Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [3]
- Immunohistochemistry [2]
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Validation data
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- Product number
- PA5-21660 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Radixin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, mouse liver. Predicted reactivity: Mouse (98%), Rat (98%), Pig (99%), Chicken (98%), Bovine (98%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.9 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 analysis of Radixin using 30 µg of 293T lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a Radixin polyclonal antibody (Product # PA5-21660) at a dilution of 1:5000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on membrane enriched cell extract (30 µg lysate) of HeLa (Lane 1), HEK-293 (Lane 2), NTERA-2 (Lane 3), COS-7 (Lane 4), K-562 (Lane 5) and Mouse Kidney (Lane 6). The blot was probed with Anti-Radixin Polyclonal Antibody (Product # PA5-21660, 1:5000 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). A 80 kDa band corresponding to Radixin was detected in cell lines tested with non-specific bands at ~50 and ~35 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Radixin Polyclonal Antibody detects RDX protein by Western blot analysis. A. 50 µg mouse liver lysate/extract.7.5 % SDS-PAGE. Radixin Polyclonal Antibody (Product # PA5-21660) dilution: 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Radixin was achieved by transfecting HeLa cells with Radixin specific siRNAs (Silencer® select Product # s11901, s11900). Western blot analysis (Fig. a) was performed using whole cell extracts from the Radixin knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Radixin Polyclonal Antibody (Product # PA5-21660, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Radixin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Radixin in paraformaldehyde-fixed HeLa cells using a Radixin polyclonal antibody (Product # PA5-21660) at a 1:200 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of paraformaldehyde-fixed HeLa, using Radixin antibody (Product # PA5-21660) at 1:200 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Radixin was performed using 70% confluent log phase HeLa cells. 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 Radixin Polyclonal Antibody (Product # PA5-21660) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled 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 ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing membrane and cytoplasmic localization. Panel e 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 RT2 xenograft, using Radixin (Product # PA5-21660) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded U373 xenograft, using Radixin (Product # PA5-21660) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.