Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [3]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-31865 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ISG15 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: HeLa, rat spleen. Predicted reactivity: Human (99%), Rhesus Monkey (90%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.45 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 ISG15 using 30 µg of HeLa lysate. Samples were loaded onto a 15% SDS-PAGE gel and probed with an ISG15 polyclonal antibody (Product # PA5-31865) at a dilution of 1:2000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of ISG15 was performed by separating 50 µg of rat tissue extract by 15% SDS-PAGE. Proteins were transferred to a membrane and probed with a ISG15 Polyclonal Antibody (Product # PA5-31865) at a dilution of 1:2,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using ISG15 Polyclonal Antibody (Product # PA5-31865). Untreated (–) and treated (+) A549 whole cell extracts (30 µg) were separated by 15% SDS-PAGE, and the membrane was blotted with ISG15 Polyclonal Antibody (Product # PA5-31865) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of A549 (Lane 1), A549 treated with IFNa (1000U/ml for 24 hours) (Lane 2), HeLa (Lane 3), HeLa treated with IFNa (1000U/ml for 24 hours) (Lane 4) and MCF7 (Lane 5). The blot was probed with Anti-ISG15 Polyclonal Antibody (Product # PA5-31865, 1:1000 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 17kDa band corresponding to ISG15 was observed across all the cell lines tested and was induced upon treatment of A549 and HeLa cells with IFNa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- ISG15 Polyclonal Antibody detects ISG15 protein at cytoplasm and nucleus by immunofluorescent analysis. Sample: HepG2 cells were fixed in 4% paraformaldehyde for 10 min. Green: ISG15 protein stained by ISG15 Polyclonal Antibody (Product # PA5-31865) diluted at 1:100. Blue: Hoechst 33342 staining. Scale bar = 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- ISG15 Polyclonal Antibody detects ISG15 protein at cytoplasm and nucleus by immunofluorescent analysis. Sample: HepG2 cells were fixed in 4% paraformaldehyde for 10 min. Green: ISG15 protein stained by ISG15 Polyclonal Antibody (Product # PA5-31865) diluted at 1:100. Blue: Hoechst 33342 staining. Scale bar = 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of ISG15 was performed using 70% confluent log phase U-937 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 ISG15 Polyclonal Antibody (Product # PA5-31865) at 1:100 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 cytoplasm and nucleus 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 human gastric cancer, using ISG15 Polyclonal Antibody (Product # PA5-31865) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.