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<strong>Figure 1 Western Blot Validation in HeLa Cell Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TWEAK 3791 (A: 1 μg/mL and B: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Western Blot Validation with Recombinant Protein</strong><br>Loading: 30 ng of human TWEAK recombinant protein per lane.Antibodies: TWEAK 3791 (Lane 1: 0.5 μg/mL, Lane 2: 1 μg/mL, Lane 3: 2 μg/mL and Lane 4: 4 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Observed at around 18kD.
<strong>Figure 3 Western Blot Validation in Mouse Pancreas Tissue Lysates</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TWEAK 3791 (Lane 1: 1 μg/mL and Lane 2: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Western Blot Validation in Mouse Liver Tissue Lysates</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TWEAK 3791 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 5 Immunohistochemistry Validation of TWEAK in Human Brain Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded human brain tissue using anti-TWEAK antibody (3791) at 10 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 6 Immunohistochemistry Validation of TWEAK in Mouse Brain Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-TWEAK antibody (3791) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 6 Immunofluorescence Validation of TWEAK in Mouse Brain Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse brain tissue labeling TWEAK with 3791 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).
Figure 1 Western Blot Validation in HeLa Cell Lysate
Loading: 15 μg of lysates per lane.Antibodies: TWEAK 3791 (A: 1 μg/mL and B: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

TWEAK Antibody

CATALOG NUMBER: 3791

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Immunogen:
Anti-TWEAK antibody (3791) was raised against recombinant human TWEAK protein.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
WB: 0.5-4 μg/mL; IHC: 5-10 μg/mL; IF: 20 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in human and mouse samples; Immunofluorescence in mouse samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1303 - Human Brain Tissue Lysate
Positive Control 2:
Cat. No. 1201 - HeLa Cell Lysate
Positive Control 3:
Cat. No. 10-301 - Human Brain Tissue Slide
Predicted Molecular Weight:
Predicted: 27 and 36 kD
Observed: 27 and 36 kD
Validation:

Recombinant Protein Test (Figure 2): Anti-TWEAK antibodies (3791) detected human TWEAK recombinant protein at different concentrations.

Isoforms:
Human TWEAK has 2 isoforms, including isoform 1 (249aa, 27kD) and isoform 2 (330aa, 36kD). Mouse TWEAK has one isoform (249aa, 27kD) and Rat TWEAK also has one isoform (249aa, 27kD).
Purification:
TWEAK Antibody is Protein A-purified.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
TWEAK Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
TWEAK antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
TNFSF12
Additional Names:
TWEAK Antibody: APO3L, DR3LG, TWEAK, APO3L, UNQ181/PRO207, Tumor necrosis factor ligand superfamily member 12, APO3 ligand
Protein Accession Number:
NP_003800
Protein Gi Number:
4507597
Ncbi Gene Id Number:
8742
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
TWEAK Antibody: TNF-related weak inducer of apoptosis (TWEAK) is a member of the tumor necrosis factor superfamily (TNFSF) of structurally related cytokines. Like most other members of this family, TWEAK is a cell surface-associated type II transmembrane protein although a smaller, biologically active form can also be generated by cleavage near the cell membrane. TWEAK has multiple biological activities, including stimulation of cell growth and angiogenesis, induction of inflammatory cytokines, in addition to stimulation of apoptosis. The TWEAK signal transduction pathway has not been well established but it appears to signal via TweakR/Fn14 in a manner similar to that described for other TNFSF members that bind receptors lacking death domains.
Background Reference 1:
Wiley and Winkles. Cyto. And Growth Factor Rev. 2003; 14:241-9.
Background Reference 2:
Chicheportiche et al. J. Biol. Chem. 1997; 272:32401-10.
Background Reference 3:
Lynch et al. J. Biol. Chem. 1999; 274:8455-9.
Background Reference 4:
Chicheportiche et al. Arthr. Res. 2002; 4:126-33. 5.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

3791

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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