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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [2]
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- 700121 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TRAF3 Recombinant Rabbit Monoclonal Antibody (12H13L59)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- This antibody is predicted to react with chimpanzee, Rhesus monkey, mouse, rat, porcine, canine, bovine and equine based on sequence homology.
- Antibody clone number
- 12H13L59
- Concentration
- 0.5 mg/mL
Submitted references A20 undermines alternative NF-κB activity and expression of anti-apoptotic genes in Helicobacter pylori infection.
Cellular inhibitors of apoptosis are global regulators of NF-κB and MAPK activation by members of the TNF family of receptors.
Lim MCC, Maubach G, Birkl-Toeglhofer AM, Haybaeck J, Vieth M, Naumann M
Cellular and molecular life sciences : CMLS 2022 Jan 28;79(2):102
Cellular and molecular life sciences : CMLS 2022 Jan 28;79(2):102
Cellular inhibitors of apoptosis are global regulators of NF-κB and MAPK activation by members of the TNF family of receptors.
Varfolomeev E, Goncharov T, Maecker H, Zobel K, Kömüves LG, Deshayes K, Vucic D
Science signaling 2012 Mar 20;5(216):ra22
Science signaling 2012 Mar 20;5(216):ra22
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TRAF3 was performed by loading 20 µg of A431 (lane1), A549 (lane2) and HeLa (lane3) cell lysates using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. TRAF3 was detected at ~63 kDa using TRAF3 Recombinant Rabbit Monoclonal Antibody (Product # 700121) at 0.5-1 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat anti-Rabbit IgG-HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TRAF3 in HeLa and A431 cell lysate using a TRAF3 recombinant rabbit monoclonal antibody (Product # 700121) at a dilution of 2 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of TRAF3 was achieved by transfecting A549 cells with TRAF3 specific siRNAs (Silencer® select Product # s14382, s14383). Western blot analysis (Fig a) was performed using whole cell extracts from the TRAF3 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti- TRAF3 Rabbit monoclonal Antibody (Product # 700121, 1µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram(Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to TRAF3.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of TRAF3 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with TRAF3 Recombinant Rabbit Monoclonal Antibody (Product # 700121) at 1 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization and panel e is a no primary antibody control. The images were captured at 20X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of TRAF3 showing staining in the cytoplasm of paraffin-embedded human brain tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a TRAF3 Monoclonal Antibody (Clone 12H13L59), Recombinant (Product # 700121) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
