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<strong>Figure 1 Western Blot Validation in Human Cell Lines </strong><br>Loading: 15 μg of lysates per lane.Antibodies: sRANK-L 3963 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 1 Western Blot Validation in Mouse Tissues</strong><br>Loading: 15 μg of lysates per lane.Antibodies: sRANK-L 3963 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Western Blot Validation with Recombinant Protein</strong><br>Loading: 30 ng of human sRANK-L recombinant protein per lane.Antibodies: sRANK-L 3963 (A: 1 μg/mL, B: 2 μg/mL and C: 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 27kD.
<strong>Figure 4 Western Blot Validation in Rat Liver Tissue</strong><br>Loading: 15 μg of lysates per lane.Antibodies: sRANK-L 3963 (A: 0.25 μg/mL and B: 0.5 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 sRANK-L in Human Liver Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded human liver tissue using anti sRANK-L antibody (3963) 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 Immunohistochemistry and Induced Expression Validation of sRANK-L in Rat Osteogenic Compartment (Hassan et al., 2017) </strong><br>Protein analysis for sRANK-L by immunohistochemistry with anti-sRANK-L antibodies (3963) in rat osteogenic cells. Strong immunoreactivity is shown for sRANK-L in osteogenic cells at 18hr after upper jaw molar extraction as compared to baseline.
Figure 1 Western Blot Validation in Human Cell Lines
Loading: 15 μg of lysates per lane.Antibodies: sRANK-L 3963 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

sRANK Ligand Antibody

CATALOG NUMBER: 3963

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Immunogen:
Anti-sRANK-L antibody (3963) was raised against a peptide corresponding to 14 amino acids near the center of human sRANK-L.
The immunogen is located within amino acids 140-190 of sRANK-L.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB
Application Note:
WB: 0.25-4 μg/mL; IHC: 5 μg/mL.
Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human and rat samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1464 - Rat Liver Tissue Lysate
Positive Control 2:
Cat. No. 1304 - Human Liver Tissue Lysate
Positive Control 3:
Cat. No. 10-201 - Human Liver Tissue Slide
Positive Control 4:
Cat. No. 1404 - Mouse Liver Tissue Lysate
Predicted Molecular Weight:
Predicted: 36kD
Observed: 36 kD
Validation:

Recombinant Protein Test (Figure 3): Anti-sRANK-L antibodies (3963) detected human sRANK-L recombinant protein at different concentrations.

Induced expression validation (Figure 6): sRANK-L expression detected by anit-sRANK-L antibodies (3963) was up-regulated after 18hr by extraction of  upper jaw molars in rat osteogenic cells.

Isoforms:
Human sRANK-L has 3 isoforms, including isoform 1 (317aa, 35.5kD), isoform 2 (244aa, 27.7kD) and isoform 3 (270aa, 30.5kD). Mouse sRANK-L also has 3 isoforms, including isoform 1 (316aa, 35kD), isoform 2 (287aa, 32.3kD) and isoform 3 (199aa, 22.2kD). Rat sRANK-L has one isoform (318aa, 38.4kD). 3963 can detect human, mouse and rat.
Purification:
sRANK Ligand Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
sRANK Ligand Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
sRANK Ligand antibody can be stored at 4°C, stable for up to one year. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
TNFSF11
Additional Names:
sRANK Ligand Antibody: ODF, OPGL, sOdf, CD254, OPTB2, RANKL, TRANCE, hRANKL2, Tumor necrosis factor ligand superfamily member 11, Osteoclast differentiation factor, ODF
Protein Accession Number:
NP_003692
Protein Gi Number:
4507595
Ncbi Gene Id Number:
8600
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
sRANK Ligand Antibody: The receptor activator of NF-κB ligand (RANK-L) is a recently discovered member of the TNF-ligand family involved in the regulation of the T cell-dependent immune response, lymph node organogenesis and bone formation. RANK-L exists as both a normal, transmembrane form and a truncated, soluble form (sRANK-L), both of which can stimulate the receptor. Activation of T cells, such as by treatment with interleukin-7, induces RANK-L production and leads to an increase of osteoclast formation and bone loss. Finally, sRANK-L can activate the antiapoptotic kinase Akt through a signaling complex involving Src kinase and TRAF6, suggesting sRANK-L may also play a role in regulating apoptosis. This antibody will recognize both the soluble form and the uncleaved transmembrane form of RANK-L.
Background Reference 1:
Wong, et al. J. Biol. Chem. 1997; 272:25190-4.
Background Reference 2:
Kong et al. Nature 1999; 397:315-23.
Background Reference 3:
Weitzmann et al. Blood 2000; 96:1873-8.
Background Reference 4:
Bharti et al. J. Biol. Chem. 2004; 279:6065-76.
Citation 1:
Hassan et al. Coordination of early cellular reactions during activation of bone resorption in the rat mandible periosteum: An immunohistochemical study. Heliyon. 2017;3(10):e00430. PMID: PMID: 29226261.

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:

3963

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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