Anti-CD45 antibody [MEM-28]

• 产品名称

  Anti-CD45抗体[MEM-28]
  参阅全部 CD45 一抗

• 描述

  小鼠单克隆抗体[MEM-28] to CD45

• 宿主

  Mouse

• 特异性

  Human CD45 antigen (LCA). This antibody reacts with all alternative forms of CD45.

• 经测试应用

  适用于: Flow Cyt, IHC-P, WB, ICC/IFmore details

• 种属反应性

  与反应: Human
  

• 免疫原

  Tissue, cells or virus corresponding to Human CD45. Human thymocytes and T lymphocytes.

• 阳性对照

• IHC-P: Human tonsil tissue. Flow Cyt: Human peripheral blood mononuclear cells. ICC/IF: Human peripheral blood mononuclear cells.

• 常规说明

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 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: 7.40
  Preservative: 0.097% Sodium azide
  Constituent: PBS

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  Protein A purified

• 克隆

  单克隆

• 克隆编号

  MEM-28

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab8216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。

• 数据库链接

• Entrez Gene: 5788 Human

• Omim: 151460 Human

• SwissProt: P08575 Human

• Unigene: 654514 Human

• 别名

• CD45 antigen antibody

• CD45R antibody

• GP180 antibody

• L-CA antibody

• LCA antibody

• Leukocyte common antigen antibody

• loc antibody

• Ly-5 antibody

• LY5 antibody

• Ly5, homolog of antibody

• Lyt-4 antibody

• OTTHUMP00000033813 antibody

• OTTHUMP00000033816 antibody

• OTTHUMP00000033817 antibody

• OTTHUMP00000038574 antibody

• Protein tyrosine phosphatase receptor type c polypeptide antibody

• Protein tyrosine phosphatase, receptor type C antibody

• protein tyrosine phosphatase, receptor type, C antibody

• Protein tyrosine phosphatase, receptor type, c polypeptide antibody

• Ptprc antibody

• PTPRC_HUMAN antibody

• Receptor-type tyrosine-protein phosphatase C antibody

• T200 antibody

• T200 glycoprotein antibody

• T200 leukocyte common antigen antibody

• B220 antibody

• CD 45 antibody

• CD45 antibody

• 

  Flow Cytometry - Anti-CD45 antibody [MEM-28] (ab8216)

  Separation of human lymphocytes (red-filled) from CD45 negative blood debris (black-dashed) in flow cytometry analysis (surface staining) of human peripheral whole blood stained using ab8216 at 2 μg/ml (GAM APC).

• 

  Western blot - Anti-CD45 antibody [MEM-28] (ab8216)

  All lanes : Anti-CD45 antibody [MEM-28] (ab8216) at 1 µg/ml
  
  Lanes 1 & 5 : Jurkat whole cell extracts with reducing SDS loading buffer
  Lane 2 : Raji whole cell extracts with reducing SDS loading buffer
  Lane 3 : HeLa whole cell extracts with reducing SDS loading buffer
  Lanes 4 & 8 : HEK293T whole cell extracts with non-reducing SDS loading buffer
  Lane 6 : Raji whole cell extracts with non-reducing SDS loading buffer
  Lane 7 : HeLa whole cell extracts with non-reducing SDS loading buffer
  
  Secondary
  All lanes : IRDye 800CW Goat-anti-Mouse IgG
  
  Predicted band size: 147 kDa
  
  
  
  

  Western blotting analysis was performed on whole cell extracts (RIPA lysis buffer) of Jurkat, Raji, HeLa, and HEK293T cell lines, mixed and heated (100°C, 5 min) with reducing and non-reducing SDS-loading buffer. Samples were resolved using 7% Tris-glycine SDS gel electrophoresis.
  
  Nitrocellulose membrane blot was probed with ab8216 (1 µg/ml), followed by IRDye 800CW Goat-anti-Mouse IgG (green). Multiplex fluorescent Western blot detection was performed.
  
  CD45 molecules were detected at ~180-250 kDa in Jurkat and Raji cell lines.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [MEM-28] (ab8216)Image from Nel, Ivonne et al. PLoS ONE 11.4 (2016): e0153018. doi: 10.1371/journal.pone.0153018 Fig 3.

  Immunocytochemistry/ Immunofluorescence analysis of CTC isolated from mRCC patients stained for hematopoietic cells labeling CD45 with ab8216 (green). The cells were fixed in 4.5% paraformaldehyde for 15 min, washed in PBS, permeabilized with 1x Perm/Wash for 10 min, washing in PBS, blocking of unspecific antibody reactions by incubation with blocking solution containing 5% BSA for 30 min, binding of Anti-CD45 antibody [MEM-28] (ab8216) (final concentration: 5 μg/ml) overnight at 4°C, wash in 0,1% Tween, binding of secondary antibody (Cy3-conjugated goat anti-mouse) for 30 min at 37°C, washing in 0,1% Tween. Subsequently, cells were stained with DAPI for 10 min, mounted with anti-fading medium and stored in the dark until evaluation. Left: CD45, right: DAPI, bottom: merge.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [MEM-28] (ab8216)

  Ab8216 staining human normal tonsil tissue. Staining is localised to cellular membranes.
  Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
  Sections were stained using an automated system (DAKO Autostainer Plus ), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

• 

  Flow Cytometry - Anti-CD45 antibody [MEM-28] (ab8216)

  Flow cytometry analysis (surface staining) of human peripheral blood cells with ab8216 (1 ug/ml), GAM-APC.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [MEM-28] (ab8216)Image from Weller, Patrick et al. PLoS ONE 9.12 (2014): e113706. doi: 10.1371/journal.pone.0113706. Fig 2. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Immunocytochemistry/ Immunofluorescence analysis of citrated peripheral blood hematologic cells taken from head and neck squamous cell carcinoma (HNSCC) patients labeling CD45 with ab8216 (green). The staining method included fixation of the cells in 4.5% paraformaldehyde for 15 min, washing in PBS, permeabilization with 1× Perm/Wash Buffer for 10 min, washing in PBS, blocking of unspecific antibody reactions by incubation with blocking solution containing 5% BSA for 30 min, binding of Anti-CD45 antibody [MEM-28] (ab8216) (final concentration: 5 µg/ml) overnight at 4°C, wash in 0,1% Tween, binding of Cy3-conjugated secondary antibody for 30 min at 37°C, washing in 0,1% Tween.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [MEM-28] (ab8216)Image from Nel, Ivonne et al. PLoS ONE 11.4 (2016): e0153018. doi: 10.1371/journal.pone.0153018 Fig 2. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Immunocytochemistry/ Immunofluorescence analysis of hematopoietic cells labeling CD45 with ab8216. The cells were fixed in 4.5% paraformaldehyde for 15 min, washed in PBS, permeabilized with 1x Perm/Wash for 10 min, washing in PBS, blocking of unspecific antibody reactions by incubation with blocking solution containing 5% BSA for 30 min, binding of Anti-CD45 antibody [MEM-28] (ab8216) (final concentration: 5 μg/ml) overnight at 4°C, wash in 0,1% Tween, binding of secondary antibody (Cy3-conjugated goat anti-mouse) for 30 min at 37°C, washing in 0,1% Tween.

• 

  Flow Cytometry - Anti-CD45 antibody [MEM-28] (ab8216)

  Overlay histogram showing peripheral blood lymphocytes stained with ab8216 (red line). The cells were incubated with the antibody (ab8216, 1µg/1x106 cells) for 30 min at 4ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H&L) (ab96879) at 1/200 dilution for 30 min at 4ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed gating on peripheral blood lymphocytes.
  
  

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [MEM-28] (ab8216)

  Immunocytochemistry/Immunofluorescence analysis of human peripheral blood mononuclear cells labelling CD45 (green) with ab8216 at 10 μg/mL. Nuclei were counterstained with DAPI (blue).