SCF Antibody (YA6373)
(Synonyms: SF; SCF; MGF; FPH2; KL-1; Kitl.)Based on 1 Customer Validation
SCF Antibody (YA6373) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to SCF.
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Host:
Rabbit
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Isotype:
IgG
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Application:
WB, IHC-P, IHC-F, ICC/IF, IF-Tissue, FC
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Reactivity :
Human
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Formulation:
Supplied in 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% glycerol.
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Conjugation:
Non-conjugated
Applications
| Application |
WB
WB: Western Blot
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IHC-P
IHC-P: Immunohistochemistry-Paraffin
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IHC-F
IHC-F: Immunohistochemistry-Frozen
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ICC/IF
ICC/IF: Immunocytochemistry/
Immunofluorescence |
ICC/IF
ICC/IF: Immunocytochemistry/
Immunofluorescence |
FC
FC: Flow Cytometry
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|---|---|---|---|---|---|---|
| Dilution Ratio | 1:500-2000 | 1:100-500 | 1:100-500 | 1:50-200 | 1:100-500 | 1:50-100 |
Product Details
SCF Antibody (YA6373) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to SCF.
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Host Rabbit
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Clonality Monoclonal
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Species ReactivityHuman
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Observed Molecular WeightObserved band size: 18 kDaNote: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
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Calculated Molecular Weight Predicted band size: 31 kDa
A synthesized peptide derived from human SCF aa 40-90/273
affinity purified.
Non-conjugated
Unmodified
IgG
Product Properties
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Appearance
Liquid
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Formulation
Supplied in 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% glycerol.
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Concentration
Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration
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Storage & Stability
Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
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Shipping
Shipping with blue ice.
Verification Images
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Western blot analysis of extracts from Raji (lane 2(20μg), HT-29 (lane 3(20μg), A549 (lane 4(20μg), MCF-7 (lane 5(20μg) using SCF Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded Human Endometrial cancer tissue using SCF Antibody ( HY-P86681, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded Human Prostate cancer tissue using SCF Antibody ( HY-P86681, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded Human Gallbladder tissue using SCF Antibody ( HY-P86681, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded Human Urinary bladder tissue using SCF Antibody ( HY-P86681, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded Human Cerebellum tissue using SCF Antibody ( HY-P86681, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded Human spleen tissue using SCF Antibody ( HY-P86681, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow Cytometry analysis of MCF-7 cells labelling SCF (red) with SCF Antibody (anti-SCF ) (HY-P86681). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then cells were stained with the primary antibody at 1/200 dilution for an hour at 4℃. Goat Anti-Rabbit IgG H&L (AF488) (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control(HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).
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Immunocytochemistry analysis of MCF-7 cells labeling SCF with SCF antibody (HY-P86681) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with SCF antibody (HY-P86681) at 1/50 dilution in BSA for Immunol Staining at 4℃,Stay overnight. AF488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L(HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue). 3.4细胞培养: 测活使用细胞株:HepG2细胞 3.5正式实验步骤 3.5.1 细胞爬片 取10μL无菌水滴于24孔板中央,随后用镊子夹取爬片覆盖于水滴之上,并用镊子按压,使爬片吸附于24孔板孔底; 细胞以6000个/孔的密度接种于爬片,每孔100μL,随后将24孔板置于37℃、5%CO2培养箱中静置6h,待细胞贴壁; 待细胞贴壁后,6孔板每孔加入1mL的完全培养基继续培养,随后将24孔板置于37℃、5%CO2培养箱中过夜培养。 3.5.2固定 将准备好的贴壁细胞上清吸去,每孔500μL 4%的多聚甲醛,室温固定15min。 3.5.3通透 吸去固定液,用PBS清洗三次,每次5分钟,随后吸弃。每孔500μL 0.1% Triton X-100通透液通透细胞膜10分钟。 3.5.4封闭 每孔500μL封闭液封闭10min; 3.5.5孵育一抗 取爬片于载玻片,封闭液稀释一抗,加入稀释后的一抗100μL,于湿盒中4度过夜孵育,第二天复温10min,随后用PBS漂洗3次,每次冲洗5min; 3.5.6二抗孵育 封闭液稀释二抗,加入稀释后的二抗100uL,室温下于湿盒中孵育1h; 3.5.7固定拍照 1) 复染核:滴加DAPI避光孵育5 min,进行染核,PBS 5 min×3次洗去多余的DAPI。 2) 用吸水纸吸干爬片上的液体,用含抗荧光淬灭剂的封片液封片,在荧光显微镜下观察采集图像 3.6 正式实验数据处理
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Immunocytochemistry analysis of HepG2 cells labeling SCF with SCF antibody (HY-P86681) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with SCF antibody (HY-P86681) at 1/50 dilution in BSA for Immunol Staining at 4 ℃,Stay overnight. AF488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L(HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
Background
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Function
SCF is a Ligand for the receptor-type protein-tyrosine kinase KIT. Plays an essential role in the regulation of cell survival and proliferation, hematopoiesis, stem cell maintenance, gametogenesis, mast cell development, migration and function, and in melanogenesis. KITLG/SCF binding can activate several signaling pathways. Promotes phosphorylation of PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase, and subsequent activation of the kinase AKT1. KITLG/SCF and KIT also transmit signals via GRB2 and activation of RAS, RAF1 and the MAP kinases MAPK1/ERK2 and/or MAPK3/ERK1. KITLG/SCF and KIT promote activation of STAT family members STAT1, STAT3 and STAT5. KITLG/SCF and KIT promote activation of PLCG1, leading to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate. KITLG/SCF acts synergistically with other cytokines, probably interleukins
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Subcellular Localization
Cell membrane; Single-pass type I membrane protein; Cytoplasm; Cytoplasm, cytoskeleton; Cell membrane; Single-pass type I membrane protein; Cell projection, lamellipodium; Cell projection, filopodium; Secreted
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Isoforms & Post-Translational Modification
P21583 has 3 isomers: P21583-1: 30899 Da (predicted); P21583-2: 27867 Da (predicted); P21583-3: 26667 Da (predicted).
A soluble form (sKITLG) is produced by proteolytic processing of isoform 1 in the extracellular domain;Found in two differentially glycosylated forms, LMW-SCF and HMW-SCF. LMW-SCF is fully N-glycosylated at Asn-145, partially N-glycosylated at Asn-90, O-glycosylated at Ser-167, Thr-168 and Thr-180, and not glycosylated at Asn-97 or Asn-118. HMW-SCF is N-glycosylated at Asn-118, Asn-90 and Asn-145, O-glycosylated at Ser-167, Thr-168 and Thr-180, and not glycosylated at Asn-97;A soluble form exists as a cleavage product of the extracellular domain -
Subunit
Homodimer, non-covalently linked (Probable). Heterotetramer with KIT, binding two KIT molecules; thereby mediates KIT dimerization and subsequent activation by autophosphorylation
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SwissProt ID
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Synonyms
SF; SCF; MGF; FPH2; KL-1; Kitl.
Documentation
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Data Sheet (234 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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User Guide for Antibodies (1077 KB)