Anti-CD14 antibody [SC69-02] 

产品描述

Lipopolysaccharide (LPS) elicits the secretion of mediators and cytokines produced by activated macrophages and monocytes. CD14 is a glycosylphosphatidylinositol (GPI)-anchored protein found on the surfaces of monocytes and polymorphonuclear leukocytes. CD14 functions as a receptor for LPS, resulting in the secretion of various proteins. An important component in the LPS activation of monocytes through the CD14 receptor is the "adapter molecule," lipopolysaccharide binding protein (LBP). There are two forms of CD14, a membrane-associated form (mCD14), and a soluble form (sCD14). mCD14 responds to LPS alone and facilitates the secretion of proteins, while cells not expressing mCD14 fail to respond to LPS. The cells that lack mCD14 respond to LPS/LBP in the presence of sCD14.

产品名称Anti-CD14 antibody [SC69-02]

分子量40 kDa

种属反应性Human,Mouse

验证应用WB, ICC, IHC-P

抗体类型重组兔单抗

免疫原Synthetic peptide within human CD14 aa 300-340.

偶联Non-conjugated

性能

形态Liquid

浓度1 mg/mL.

存放说明Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

存储缓冲液1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型IgG

纯化方式Protein A affinity purified.

亚细胞定位Cell membrane, Secreted, Golgi apparatus, Membrane raft.

数据链接SwissProt: P08571 Human

其它名称

CD 14 antibody

CD_antigen=CD14 antibody

CD14 antibody

more

应用

• WB: 1:500-1:2,000
  ICC: 1:50-1:200
  IHC-P: 1:50-1:200

• 

  Fig1: Western blot analysis of CD14 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  Positive control:
  Lane 1: human liver tissue lysate
  Lane 2: SW480 cell lysate

  

  Fig2: ICC staining of CD14 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibodyfor 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig3: ICC staining of CD14 in NCCIT cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig4: ICC staining of CD14 in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig5: ICC staining of CD14 in LO2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody  for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig6: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD14 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody for 30 minutes at room temperature. 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.

  

  Fig7: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-CD14 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody ( for 30 minutes at room temperature. 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.

  

  Fig8: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-CD14 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody  for 30 minutes at room temperature. 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.

  

  Fig9: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CD14 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody for 30 minutes at room temperature. 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.

  

  Fig10: Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-CD14 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody ( for 30 minutes at room temperature. 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.