SCF Antibody (YA6373)

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.

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.

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.

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.

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.

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.

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.

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).

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

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).