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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Flow cytometry [1]
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- Product number
- MA5-16164 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TNFAIP3 Monoclonal Antibody (59A426)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 59A426
- Vial size
- 100 µg
- Concentration
- 1.0 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with IL1α (20 ng/mL for 4 hours) (Lane 2), HeLa treated with TNFα (20 ng/mL for 12 hours) (Lane 3), THP1 (Lane 4), THP1 treated with PMA (80 nM for 12 hours) (Lane 5) and PMA differentiated THP1 macrophages treated with TNFα (20 ng/mL for 12 hours) (Lane 6). The blot was probed with Anti-TNFAIP3 Monoclonal Antibody (Product # MA5-16164, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 90 kDa band corresponding to TNFAIP3 was observed to be induced upon IL1α or TNFα treatment of HeLa (Lanes 1-3) or TNFα treatment of PMA differentiated THP1 macrophages (Lanes 4-6).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TNFAIP3 in Human Jurkat lysate. Samples were incubated in TNFAIP3 monoclonal antibody (Product # MA5-16164) using a dilution of 4 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TNFAIP3 in 0.05 mg/mL Jurkat lysate. Samples were incubated in TNFAIP3 monoclonal antibody (Product # MA5-16164). This experiment was performed under reducing conditions using the 12-230 kDa separation system..
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of TNFAIP3 was performed using 70% confluent log phase RAW 264.7 cells treated with 1 µg/mL of LPS for 6 hr. 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 TNFAIP3 Mouse Monoclonal Antibody (Product # MA5-16164) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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, 1:300). Panel d represents the merged image showing cytosolic localization. Panel e shows untreated cells with less cytosolic signal. 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 TNFAIP3 in Human placenta. Samples were incubated with TNFAIP3 monoclonal antibody (Product # MA5-16164) using a dilution of 5 µg/mL. Cytoplasmic staining of decidual cells is observed. Human tissue TMA was used for this test. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of A20 in ThP-1 cells using a A20/TNFAIP3 monoclonal antibody (Product # MA5-16164) (red) and an isotype control (green) at 0.5 µg/mL.
