Anti-CD3 antibody [CD3-12]

• 产品名称

  Anti-CD3抗体[CD3-12]
  参阅全部 CD3 一抗

• 描述

  大鼠单克隆抗体[CD3-12] to CD3

• 宿主

  Rat

• 特异性

  IHC-P is not recommended in Human samples.

• 经测试应用

  适用于: IHC-P, WB, Flow Cyt (Intra)more details

• 种属反应性

  与反应: Mouse, Rat, Human
  

• 免疫原

  Synthetic peptide corresponding to Human CD3. Derived from cytoplasmic epitope.
  Database link: P07766

  Run BLAST with Run BLAST with 

• 阳性对照

• IHC-P: Mouse spleen and lymph node tissue; Rat spleen tissue. Flow Cyt (Intra): Human peripheral blood lymphocytes. WB: Jurkat, MOLT-4 and EL4 cell lysates; Human, mouse and rat thymus tissue lysates.

• 常规说明

  This product has switched from a hybridoma to recombinant production method on 1st November 2024.

  This product is a recombinant monoclonal antibody, which offers several advantages including:

  For more information see here.

• - High batch-to-batch consistency and reproducibility

• - Improved sensitivity and specificity

• - Long-term security of supply

• - Animal-free production

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

• 存储溶液

  pH: 7.20
  Preservative: 0.01% Sodium azide
  Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  Protein A purified

• 克隆

  单克隆

• 克隆编号

  CD3-12

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab11089于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 

  Flow Cytometry - Anti-CD3 antibody [CD3-12] (ab11089)

  Flow cytometry analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized Human peripheral blood mononuclear cells (PBMC) stained with ab11089 at 1/500 dilution and Goat F(ab)2 Anti-Rat IgG Fc (Alexa Fluor® 488, ab150161) at 1/2000 dilution. Isotype control was Rat monoclonal IgG / Left.

• 

  Western blot - Anti-CD3 antibody [CD3-12] (ab11089)

  All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1 µg/ml
  
  Lane 1 : THP-1 (Human monocytic leukemia cell line) whole cell lysate (negative control)
  Lane 2 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate (negative control)
  Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
  Lane 4 : Human thymus tissue lysate
  Lane 5 : Mouse thymus tissue lysate
  Lane 6 : Rat thymus tissue lysate
  
  Lysates/proteins at 15 µg per lane.
  
  Secondary
  All lanes : Goat anti-Rat at 1/10000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 23.1 kDa
  Observed band size: 23 kDawhy is the actual band size different from the predicted?
  
  
  
  

  This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab11089 overnight at 4°C. Antibody binding was detected using Goat anti-Rat secondary at a 1:10000 dilution for 1hr at room temperature and then imaged.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] (ab11089)

  Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CB3 with ab11089 at 1/200 dilution, followed by ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). Positive staining on rat spleen tissue is observed. Counterstained with hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] (ab11089)

  Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3 with ab11089 at 1/200 dilution, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on mouse spleen tissue is observed. Counterstained with hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

   

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] (ab11089)

  IHC image of CD3 staining in a formalin fixed, paraffin embedded normal mouse lymph node tissue.

  The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab11089 at 1/250 dilution for 15 minutes at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] (ab11089)This image is courtesy of an anonymous Abreview.

  ab11089 staining CD3 in mouse lymph node tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

  Tissue was fixed with formaldehyde and blocked with 15% serum for 60 minutes at 20°C. Antigen retrieval was by heat mediation in sodium citrate (pH 6). Samples were incubated with primary antibody (1/250 in TBS) for 18 hours at 20°C. An Alexa Fluor®conjugated goat anti-rat IgG polyclonal (1/400) was used as the secondary antibody.

  See Abreview

• 

  Western blot - Anti-CD3 antibody [CD3-12] (ab11089)

  All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1/1000 dilution
  
  Lane 1 : Mouse thymus tissue lysate
  Lane 2 : Rat thymus tissue lysate
  Lane 3 : Human thymus tissue lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution
  
  Predicted band size: 23.1 kDa
  
  
  
  

  Blocking and dilution buffer: 5% NFDM/TBST.

  Exposure: Lanes 1/3: 5 secs; Lanes 2: 3 secs.

• 

  Western blot - Anti-CD3 antibody [CD3-12] (ab11089)

  All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1/1000 dilution
  
  Lane 1 : Jurkat (human T cell leukemia T lymphocyte)
  Lane 2 : MOLT-4 (human lymphoblastic leukemia T lymphoblast)
  Lane 3 : EL4 (mouse lymphoma T lymphocyte) whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution
  
  Predicted band size: 23.1 kDa
  
  
  
  

  Blocking and dilution buffer: 5% NFDM/TBST.

  Exposure time: 5 secs.

• 

  Western blot - Anti-CD3 antibody [CD3-12] (ab11089)

  Anti-CD3 antibody [CD3-12] (ab11089) + Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
  
  Secondary
  HRP-conjugated goat anti-rat IgG
  
  Predicted band size: 23.1 kDa