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<strong>Western Blot Validation in Cell Lines and Tissues of Human, Mouse and Rat</strong><br>Loading: 15 μg of lysates per lane.Antibodies: SCF 5165 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Western Blot Validation with Recombinant Protein</strong><br>Loading: 30 ng of human SCF recombinant protein per lane.Antibodies: SCF 5165 (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.Observed at around 20kD.
<strong> Immunohistochemistry Validation of SCF in Mouse Brain Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-SCF antibody (5165) at 2.5 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>Immunofluorescence Validation of SCF in Human Brain Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed human brain tissue labeling SCF with 5165 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Regulation Validation of SCF in Streptozotocin (STZ)-induced Diabetic Mice (Zhang et al., 2016) </strong><br>WB analysis showed protein expression level of SCF detected by anti-SCF antibody (5165) in gastric smooth muscle was significantly decreased in STZ-induced diabetic mice as compared to the control.
<strong>Regulated Expression Validation of SCF in Gastric Smooth Muscle Cells (GSMCs) of Mice (Zhang et al., 2016) </strong><br>WB analysis of protein expression level of SCF detected by anti-SCF antibody (5165). GSMCs were treated with PI3K inhibitor (LY294002), AT1R inhibitor (ZD7155) and AT2R inhibitor (PD123319) before Ang II treatment. Ang II (10<sup>-8</sup>mol/L) significantly increased SCF protein expression, which was reduced by treatment of LY294002 and ZD7155.
<strong>Regulated Expression Validation of SCF in GSMCs of Normal Mice (Wu et al., 2013) </strong><br>WB analysis of protein expression level of SCF detected by anti-SCF antibody (5165). GSMCs were treated with C-type natriuretic peptide (CNP) at different doses for 48hr. CNP(10<sup>-7</sup>mol/L and 10<sup>-6</sup>mol/L) significantly decreased SCF protein expression level as compared to the control group.
Western Blot Validation in Cell Lines and Tissues of Human, Mouse and Rat
Loading: 15 μg of lysates per lane.Antibodies: SCF 5165 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

SCF Antibody

CATALOG NUMBER: 5165

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Homology:
Predicted species reactivity based on immunogen sequence: Bovine: (71%)
Immunogen:
Anti-SCF antibody (5165) was raised against a peptide corresponding to 18 amino acids near the center of human SCF.
The immunogen is located within amino acids 100-150 of SCF.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
WB: 1-2 μg/mL; IHC: 2.5 μg/mL; IF: 20 μg/mL.
Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in mouse samples; Immunofluorescence in human samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1202 – A431 Cell Lysate
Positive Control 2:
Cat. No. 10-301 - Human Brain Tissue Slide
Predicted Molecular Weight:
Predicted: 31kD
Observed: 36 kD (31kD + 4 N-linked Glycosylations)
Validation:

Recombinant Protein Test (Figure 2): Anti-SCF antibodies (5165) detected human SCF recombinant protein at different concentrations.

Regulated expression validation (Figure 7 & 8): SCF expression detected by anti-SCF antibodies (5165) was up-regulated by angiotensin II (Ang II) treatment and was down-regulated by two inhibitors (Figure 7) or CNP treatment (Figure 8).

Isoforms:
Human SCF has 3 isoforms, including isoform 1 (273aa, 30.9kD), isoform 2 (245aa, 27.9kD) and isoform 3 (238aa, 26.7kD). Mouse SCF has 2 isoforms, including isoform 1 (273aa, 30.6kD) and isoform 2 (245aa, 27.6kD). Rat SCF also has 2 isoforms, including isoform 1 (273aa, 30.7kD) and isoform 2 (245aa, 27.7kD). 5165 can detect human and possibly detect mouse and rat.
Purification:
SCF Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
SCF Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
SCF 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:
KITLG
Additional Names:
SCF Antibody: SF, MGF, SCF, FPH2, KL-1, Kitl, SHEP7, Kit ligand, Mast cell growth factor
Protein Accession Number:
P21583
Protein Gi Number:
134289
Ncbi Gene Id Number:
4254
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
SCF Antibody: Stem cell factor (SCF) is the ligand of the c-Kit oncogene and is expressed by various structural and inflammatory cells in the airways. Binding of SCF by the c-Kit receptor leads to homodimerization of the receptor and the activation of signalling pathways such as PI-3, PLC-gamma, Jak/STAT, and MAP kinase pathways. SCF expression leads to the induction of mast cell survival and the expression and release of histamine, pro-inflammatory cytokines and chemokines. The inhibition of the SCF/c-Kit pathway leads to a decrease in histamine levels, mast cell and eosinophil infiltration, IL-4 production and airway hyperresponsiveness, suggesting this pathway may be a useful therapeutic target in inflammatory diseases such as asthma. At least two isoforms of SCF are known to exist.
Background Reference 1:
Reber et al. Euro. J. Pharmacology 2006; 533:327-40.
Background Reference 2:
Jensen et al. Inflamm. Allergy Drug Targets 2007; 6:57-62.
Background Reference 3:
Lukacs et al. J. Immunol. 1996; 156:3945-51.
Citation 1:
Zhang et al. Up-regulation of the Ang II/AT1 receptor may compensate for the loss of gastric antrum ICC via the PI3k/Akt signaling pathway in STZ-induced diabetic mice. Mol Cell Endocrinol. 2016;423:77-86.PMID: 26773730
Citation 2:
Wu et al. Diabetes-induced loss of gastric ICC accompanied by up-regulation of natriuretic peptide signaling pathways in STZ-induced diabetic mice. Peptides. 2013;40:104-11. PMID: 23352981

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:

5165

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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