RANKL Antibody - #AF0313

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Product: RANKL Antibody
Catalog: AF0313
Description: Rabbit polyclonal antibody to RANKL
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 40kDa; 35kD(Calculated).
Uniprot: O14788
RRID: AB_2833477

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 100ul $280 In stock
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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:3000, IHC 1:50-1:200, IF/ICC 1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(90%), Dog(100%)
Clonality:
Polyclonal
Specificity:
RANKL Antibody detects endogenous levels of total RANKL.
RRID:
AB_2833477
Cite Format: Affinity Biosciences Cat# AF0313, RRID:AB_2833477.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

CD254; hRANKL2; ODF; OPGL; OPTB2; Osteoclast differentiation factor; Osteoprotegerin ligand; RANKL; Receptor activator of nuclear factor kappa B ligand; Receptor activator of nuclear factor kappa-B ligand; sOdf; TNF related activation induced cytokine; TNF-related activation-induced cytokine; TNF11_HUMAN; TNFSF 11; Tnfsf11; TRANCE; Tumor necrosis factor (ligand) superfamily member 11; Tumor necrosis factor ligand superfamily member 11; Tumor necrosis factor ligand superfamily member 11, soluble form;

Immunogens

Immunogen:
Uniprot:

O14788(TNF11_HUMAN) >>Visit HPA database.

Gene(ID):
TNFSF11(8600)
Expression:
O14788 TNF11_HUMAN:

Highest in the peripheral lymph nodes, weak in spleen, peripheral blood Leukocytes, bone marrow, heart, placenta, skeletal muscle, stomach and thyroid.

Description:
TNFSF11 Cytokine that binds to TNFRSF11B/OPG and to TNFRSF11A/RANK. Osteoclast differentiation and activation factor. Augments the ability of dendritic cells to stimulate naive T-cell proliferation. May be an important regulator of interactions between T-cells and dendritic cells and may play a role in the regulation of the T-cell-dependent immune response. May also play an important role in enhanced bone-resorption in humoral hypercalcemia of malignancy.
Sequence:
MRRASRDYTKYLRGSEEMGGGPGAPHEGPLHAPPPPAPHQPPAASRSMFVALLGLGLGQVVCSVALFFYFRAQMDPNRISEDGTHCIYRILRLHENADFQDTTLESQDTKLIPDSCRRIKQAFQGAVQKELQHIVGSQHIRAEKAMVDGSWLDLAKRSKLEAQPFAHLTINATDIPSGSHKVSLSSWYHDRGWAKISNMTFSNGKLIVNQDGFYYLYANICFRHHETSGDLATEYLQLMVYVTKTSIKIPSSHTLMKGGSTKYWSGNSEFHFYSINVGGFFKLRSGEEISIEVSNPSLLDPDQDATYFGAFKVRDID

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Rabbit
90
Chicken
60
Xenopus
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O14788 As Substrate

Site PTM Type Enzyme
S137 Phosphorylation

Research Backgrounds

Function:

Cytokine that binds to TNFRSF11B/OPG and to TNFRSF11A/RANK. Osteoclast differentiation and activation factor. Augments the ability of dendritic cells to stimulate naive T-cell proliferation. May be an important regulator of interactions between T-cells and dendritic cells and may play a role in the regulation of the T-cell-dependent immune response. May also play an important role in enhanced bone-resorption in humoral hypercalcemia of malignancy. Induces osteoclastogenesis by activating multiple signaling pathways in osteoclast precursor cells, chief among which is induction of long lasting oscillations in the intracellular concentration of Ca (2+) resulting in the activation of NFATC1, which translocates to the nucleus and induces osteoclast-specific gene transcription to allow differentiation of osteoclasts. During osteoclast differentiation, in a TMEM64 and ATP2A2-dependent manner induces activation of CREB1 and mitochondrial ROS generation necessary for proper osteoclast generation (By similarity).

PTMs:

The soluble form of isoform 1 derives from the membrane form by proteolytic processing (By similarity). The cleavage may be catalyzed by ADAM17.

Subcellular Location:

Cell membrane>Single-pass type II membrane protein.

Cell membrane>Single-pass type II membrane protein.

Cytoplasm.

Secreted.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Highest in the peripheral lymph nodes, weak in spleen, peripheral blood Leukocytes, bone marrow, heart, placenta, skeletal muscle, stomach and thyroid.

Subunit Structure:

Homotrimer (By similarity). Interacts with TNFRSF11B. Interacts with TNFRSF11A. Interacts with FBN1 (via N-terminal domain) in a Ca(+2)-dependent manner (By similarity).

Family&Domains:

Belongs to the tumor necrosis factor family.

Research Fields

· Environmental Information Processing > Signaling molecules and interaction > Cytokine-cytokine receptor interaction.   (View pathway)

· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.   (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

· Human Diseases > Immune diseases > Rheumatoid arthritis.

· Organismal Systems > Development > Osteoclast differentiation.   (View pathway)

· Organismal Systems > Endocrine system > Prolactin signaling pathway.   (View pathway)

References

1). IGFBP7 acts as a negative regulator of RANKL-induced osteoclastogenesis and oestrogen deficiency-induced bone loss. CELL PROLIFERATION, 2020 (PubMed: 31889368) [IF=8.5]

Application: IF/ICC    Species: mice    Sample: RAW264.7 cells

FIGURE 1 IGFBP7 suppressed RANKL-induced osteoclastogenesis in vitro. A-B, CCK-8 analysis was conducted to evaluate cell viability after treated by different concentration of recombinant IGFBP7 protein for 48 and 96 h. C, BMMs were cultured with different concentration of recombinant IGFBP7 protein in osteoclastogenic medium for 5 d, and TRAP staining was performed. Scale bar = 200 µm. D-E, The number and area of osteoclasts were calculated. F-H, BMMs were treated with either vehicle or 1000 ng/mL recombinant IGFBP7 protein from days 1-3 (early stage), days 3-5 (late stage) or days 1-5 (early + late stage) during osteoclastogenesis. The number and area of osteoclasts were calculated. Scale bar = 200 µm. I and K, Representative fluorescence images showed that recombinant IGFBP7 treatment significantly decreased the size of F-actin ring structures. Scale bar = 100 µm. J and L, Representative SEM (scanning electron microscopy) images showed that recombinant IGFBP7 treatment decreased the area of bone resorption pits. Scale bars = 200 µm. *P < .05, **P < .01 vs the control group

Application: IF/ICC    Species: mouse    Sample: osteoblastic cel

FIGURE 1 |IGFBP7 suppressed RANKL-induced osteoclastogenesis in vitro.I and K, Representative fluorescence images showed that recombinant IGFBP7 treatment significantly decreased the size of F-actin ring structures. Scale bar = 100 µm. J and L, Representative SEM (scanning electron microscopy) images showed that recombinant IGFBP7 treatment decreased the area of bone resorption pits. Scale bars = 200 µm. *P < .05, **P < .01 vs the control group

Application: WB    Species: mouse    Sample: MC3T3-E1

Figure S1.| The results of western-blot analysis showed that different concentrations of recombinant IGFBP7 increased the protein expression of OPG in osteoblastic cell line MC3T3-E1, whereas the expression of RANKL was not affected after 3 days’ IGFBP7 treatment. OPG, osteoprotegerin; RANKL, receptor activator of nuclear factor-κB ligand.
2). Mitigation of inflammatory bowel disease-related osteoporosis by oxyberberine: Insights into the RANKL/NF-κB signaling pathway. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 2024 (PubMed: 38574627) [IF=7.5]

Application: WB    Species: Mouse    Sample:

Fig. 8. OBB inhibited the RANKL/NF-κB signaling pathway in DSS-induced IBD mice. (A) Representative bands of RANKL, NFATc1, pIκBα, IκBα, cytoplasmic p65, and nuclear p65. (B-F) Relative protein expression levels of RANKL (F-value: 28.501), NFATc1 (F-value: 28.705), pIκBα, IκBα (pIκBα/IκBα, F-value: 10.056), cytoplasmic p65 (F-value:5.692), and nuclear p65 (F-value: 6.893). Data presented as mean ± SD (n = 3), degrees of freedom between groups: 4, degrees of freedom within groups
3). Yunvjian decoction attenuates lipopolysaccharide-induced periodontitis by suppressing NFκB/NLRP3/IL-1β pathway. Journal of ethnopharmacology, 2024 (PubMed: 37802377) [IF=5.4]
4). Preventive and Therapeutic Potential of Streptococcus cristatus CA119 in Experimental Periodontitis in Rats. Probiotics and antimicrobial proteins, 2024 (PubMed: 38607584) [IF=4.9]
5). Lanthanum Hydroxide and Chronic Kidney Disease Mineral and Bone Disorder: A Rat Model. Current vascular pharmacology, 2024 (PubMed: 37961858) [IF=4.5]
6). Xuetongsu attenuates bone destruction in collagen-induced arthritis mice by inhibiting osteoclast differentiation and promoting osteoclast apoptosis. The international journal of biochemistry & cell biology, 2024 (PubMed: 38340949) [IF=4.0]
7). Role of microRNA-19b-3p on osteoporosis after experimental spinal cord injury in rats. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2022 (PubMed: 33587904) [IF=3.9]

Application: WB    Species: Rat    Sample: osteoblasts

Figure 2. MiR-19b-3p inhibits osteoblast differentiation. (A) RT-PCR analysis was used to detect miRNA-19b-3p expression levels in BMSC-derived osteoblasts after miR-19b-3p-overexpressed or knockdown by corresponding lentivirus. (B) ALP activity in BMSC-derived osteoblasts after miRNA-19b-3p overexpression or knockdown by corresponding lentivirus. (C) RT-PCR was used to detect EBF2 mRNA expression levels in Journal Pre-proof 20 BMSC-derived osteoblasts after miRNA-19b-3p overexpression or knockdown by corresponding lentivirus. Western blot assay was used to detect (D and E) EBF2, (F-H) RANKL and OPG protein expression levels in BMSC-derived osteoblasts after miRNA-19b-3p overexpression or knockdown by corresponding lentivirus. (I) The ratio of OPG to RANKL in BMSC-derived osteoblasts after miRNA-19b-3p overexpression or knockdown by corresponding lentivirus (J) Alizarin red staining was used to detect cell mineralization at day 21 after differentiation after miRNA-19b-3p overexpression or knockdown at 100×. Data are analyzed using unpaired Student’s t-tests and presented as the mean ± SD from three independent experiments. * p < 0.05; ** p < 0.01 vs. the indicated group.
8). Nirogacestat suppresses RANKL-Induced osteoclast formation in vitro and attenuates LPS-Induced bone resorption in vivo. EXPERIMENTAL CELL RESEARCH, 2019 (PubMed: 31211955) [IF=3.7]

Application: IHC    Species: mouse    Sample: BMMs

Fig. 7.| Histological analysis of the inhibitory effect of PF on LPS-induced bone resorption. (A–C) Representative images of HE and TRAP staining, showing the reduced osteolytic legion and TRAP-positive OCs in the PF-treated groups. (D–G) Representative images of IHC staining of RANKL, OPG, OCN and TNF-α. (H) TRAPpositive OCs number. (H) Quantitative analysis of the expression of RANKL, OPG and RANKL/OPG ratio. The data are presented as the mean ± SD (*p < 0.05,**p < 0.01, ***p < 0.001).
9). Effects of a calcium/vitamin D/Zinc combination on anti-osteoporosis in ovariectomized rats. JOURNAL OF TRACE ELEMENTS IN MEDICINE AND BIOLOGY, 2023 (PubMed: 36773556) [IF=3.5]
10). Rodent incisor and molar dental follicles show distinct characteristics in tooth eruption. ARCHIVES OF ORAL BIOLOGY, 2021 (PubMed: 33845260) [IF=3.0]

Application: WB    Species: Rat    Sample: Incisor dental follicle (IF) cells and molar dental follicle (MF) cells

Fig. 5. Differential expression patterns of tooth eruption-related genes and proteins between IF cells and MF cells. (A) Non-induced IF cells and MF cells showed different gene expression patterns; Data are mean ± SD of n = 3 replicates, one-way ANOVA, *p＜0.05, **p＜0.01, ***p＜0.001, ****p＜0.0001. (B) Non-induced and induced IF cells and MF cells showed different protein expression patterns. After induction with CLCCM and HERSCM, there were no significant changes of the expression patterns of tooth eruption-related proteins in IF cells and MF cells. (C) The grey value ratios of Western blotting results; Data are mean ± SD of n = 3 replicates, one-way ANOVA followed by Tukey post hoc test, *p＜0.05, **p＜0.01, ***p＜0.001, ****p＜0.0001. (IF1, MF1 represent the cells cultured by α-MEM; IF2, MF2 represent the cells cultured by α-MEM + CLCCM; IF3, MF3 represent the cells cultured by α-MEM+HERSCM).

Application: WB    Species: Rat    Sample: IF cells and MF cells

Fig. 5. Differential expression patterns of tooth eruption-related genes and proteins between IF cells and MF cells. (A) Non-induced IF cells and MF cells showed different gene expression patterns; Data are mean ± SD of n = 3 replicates, one-way ANOVA, *p＜0.05, **p＜0.01, ***p＜0.001, ****p＜0.0001. (B) Non-induced and induced IF cells and MF cells showed different protein expression patterns. After induction with CLCCM and HERSCM, there were no significant changes of the expression patterns of tooth eruption-related proteins in IF cells and MF cells. (C) The grey value ratios of Western blotting results; Data are mean ± SD of n = 3 replicates, one-way ANOVA followed by Tukey post hoc test, *p＜0.05, **p＜0.01, ***p＜0.001, ****p＜0.0001. (IF1, MF1 represent the cells cultured by α-MEM; IF2, MF2 represent the cells cultured by α-MEM + CLCCM; IF3, MF3 represent the cells cultured by α-MEM+HERSCM).
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