- SynonymKITLG,FPH2,KL-1,Kitl,MGF,SCF,SF,SHEP7,KL
- SourceBiotinylated Human SCF, Avitag,His Tag (SCF-H82E1) is expressed from human 293 cells (HEK293). It contains AA Glu 26 - Ala 190 (Accession # AAH69797).Predicted N-terminus: Glu 26Request for sequence
- Molecular Characterization
This protein carries an Avi tag (Avitag™) at the C-terminus, followed by a polyhistidine tag.
The protein has a calculated MW of 21.7 kDa. The protein migrates as 26-38 kDa under reducing (R) condition (SDS-PAGE) due to different glycosylation.
- BiotinylationBiotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
- Biotin:Protein RatioThe biotin to protein ratio is 0.5-1 as determined by the HABA assay.
- EndotoxinLess than 1.0 EU per μg by the LAL method.
- Purity
>95% as determined by SDS-PAGE.
- Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
- Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
- Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human SCF, Avitag,His Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
Immobilized Human CD117, Fc Tag (Cat. No. CD7-H5255) at 2 μg/mL (100 μL/well) can bind Biotinylated Human SCF, Avitag,His Tag (Cat. No. SCF-H82E1) with a linear range of 0.1-3 ng/mL (QC tested).
- BackgroundKit ligand (KITLG) is also known as stem cell factor (SCF), mast cell growth factor (MGF), steel factor (SF), which belongs to the SCF family, and is a widely expressed 28 - 40 kDa type I transmembrane glycoprotein. KITLG is the ligand for the receptor-type protein-tyrosine kinase KIT. SCF / MGF 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. KITLG / SF 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.
- References
- (1)Anderson DM, et al., 1991, Cell Growth Differ. 2 (8): 373–8.
- (2)Broudy VC, 1997, Blood 90 (4): 1345–64.
- (3)Blouin R, Bernstein A, 1993, In Freedman MH, Feig SA.Boca Raton: CRC Press.
- (4)Kent D, et al., 2008, Clin. Cancer Res. 14 (7): 1926–30.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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(2)荧光标记法 : 使用二乙酸荧光素(FDP)、碘化丙啶(PI)或异硫氰酸荧光素钠标记的荧光染料与细胞共孵育,用流式细胞仪检测荧光染色阳性细胞的比率。此法其实是(1)法的“荧光”版,但其在灵敏性和准确性方面明显要优于后者。
(3)硝酸镧(La)示踪法: 在正常的生物组织中镧微粒可沉积于细胞间隙,但不能穿过具有1~ 2nm 微小间隙的细胞膜性结构(包括细胞膜和细胞器膜),也不能穿过细胞间的紧密连接。在膜性结构通透性增高时, 镧微粒则可进入细胞、细胞器和紧密连接内, 并在电镜下显示, 镧盐标记技术被认为是一种有效的监测细胞膜通透性变化的标记技术。
(4)LDH释放法: 在正常情况下,细胞内大分子物质LDH 是不能通过细胞膜的, 但在细胞膜受损伤而通透性增加时,可通过受损的细胞膜释放出来。LDH 能较好地反映细胞膜损伤程度。类似的还有检测细胞外K+的漏出率等。
有几个疑问
1:荧光标记到细胞是标记到细胞表面还是细胞质内?
2:荧光应该随着细胞的分化和增殖逐渐消失?是不是分化增殖越快,荧光消失速度越快?
3:有哪些容易操作,成本便宜的荧光物质?
谢谢各位战友
比如你用的CD1a-FITC(如果是鼠单抗IgG1,那对照抗体就要用相同物种的非特异性IgG1-FITC)。注意浓度要相同。一般提供抗体的公司BD,santa cruz等有提供的。其他就按照说明书的推荐浓度和孵育时间。