Anti-CD45 抗体 [MEM-28] (ab8216)

Reviews (1)Q&A (8)References (25)

Key features and details

  • Mouse monoclonal [MEM-28] to CD45
  • Suitable for: Flow Cyt, IP, IHC-P, WB, ICC/IF
  • Reacts with: Human
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-CD45 antibody [MEM-28]
    CD45 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [MEM-28] to CD45

  • 由来種

    Mouse

  • 特異性

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

  • アプリケーション

    適用あり: Flow Cyt, IP, IHC-P, WB, ICC/IFmore details

  • 種交差性

    交差種: Human

  • 免疫原

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

  • ポジティブ･コントロール

    • Jurkat human leukemia T-cell lysate; Kg-1a human leukemia cell lysate, Human peripheral blood mononuclear cells

アプリケーション

Our Abpromise guarantee covers the use of ab8216 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Flow Cyt Use 1µg for 106 cells.

(Recognizes an extracellular epitope).
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml. Antigen retrieval is not essential but may optimise staining.
WB Use a concentration of 1 µg/ml.

Prepare and run samples in non-reducing conditions.
ICC/IF Use a concentration of 10 µg/ml.

PubMed: 23908685

ターゲット情報

  • 機能

    Protein tyrosine-protein phosphatase required for T-cell activation through the antigen receptor. Acts as a positive regulator of T-cell coactivation upon binding to DPP4. The first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Upon T-cell activation, recruits and dephosphorylates SKAP1 and FYN.

  • 関連疾患

    Defects in PTPRC are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)B(+)NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development.
    Genetic variations in PTPRC are involved in multiple sclerosis susceptibility (MS) [MIM:126200]. MS is a neurodegenerative disorder characterized by the gradual accumulation of focal plaques of demyelination particularly in the periventricular areas of the brain. Peripheral nerves are not affected. Onset usually in third or fourth decade with intermittent progression over an extended period. The cause is still uncertain.

  • 配列類似性

    Belongs to the protein-tyrosine phosphatase family. Receptor class 1/6 subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 2 tyrosine-protein phosphatase domains.

  • ドメイン

    The first PTPase domain interacts with SKAP1.

  • 翻訳後修飾

    Heavily N- and O-glycosylated.

  • 細胞内局在

    Membrane. Membrane raft. Colocalized with DPP4 in membrane rafts.
  • Information by UniProt

  • 参照データベース

    • 別名

      • B220 antibody
      • CD 45 antibody
      • CD45 antibody
      • 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
      see all

    画像

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

    参考文献 (25)

    ab8216 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab8216 は 25 報の論文で使用されています。

    • Furie R  et al. Monoclonal antibody targeting BDCA2 ameliorates skin lesions in systemic lupus erythematosus. J Clin Invest 129:1359-1371 (2019). PubMed: 30645203
    • Lim WM  et al. CLIP-170 is essential for MTOC repositioning during T cell activation by regulating dynein localisation on the cell surface. Sci Rep 8:17447 (2018). PubMed: 30487641
    • Huo Z  et al. The relationship between allergic status and adenotonsillar regrowth: a retrospective research on children after adenotonsillectomy. Sci Rep 7:46615 (2017). PubMed: 28418014
    • Jackson MV & Krasnodembskaya AD Analysis of Mitochondrial Transfer in Direct Co-cultures of Human Monocyte-derived Macrophages (MDM) and Mesenchymal Stem Cells (MSC). Bio Protoc 7:N/A (2017). PubMed: 28534038
    • Jia Z  et al. Comparison of biological characteristics of nucleus pulposus mesenchymal stem cells derived from non-degenerative and degenerative human nucleus pulposus. Exp Ther Med 13:3574-3580 (2017). PubMed: 28588682
    View all Publications for this product

    レビューと Q&A

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    1-9 of 9 Abreviews or Q&A

    Question

    Hi,



    I am wondering how many biotin molecules are conjugated to each antibody.

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    Answer

    Thank you for contacting us.
    There are 3-8 molecules of biotin at each antibody molecule.
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
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    Can’t find the antibody you need? Try our Custom RabMAb® services.
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    Question


    Thanks for your reply. What is the difference when some antibodies say
    tested for ICC/IF and some just say ICC? I am using sections of a
    paraffin-embedded sample, incubating them with the primary antibodies
    I mentioned, using a fluorescent-tagged secondary antibody, mounting
    the samples, and then using a fluorescence microscope to image the
    samples.

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    Answer


    IF by definition refers to any antibody application that utilizes fluorescence as the detection method. Thus, technically ICC and IF are one in the same, and you could argue that IHC is also a form of IF.

    For the purposes of searching for products on our website, we distinguish those antibodies that have been tested in IHC with IHC followed by a letter designation (P-paraffin embedded sections, Fr-frozen sections, FoFr-formaldehyde fixed frozen sections). If an antibody has ICC or ICC/IF listed under tested applications they are the same thing and guaranteed to work in immunocytochemistry.

    It sounds like you are performing IHC-P, immunohistochemistry on paraffin embedded sections; thus, all three antibodies that you identified, ab53003, ab8216, and ab111250 are tested and guaranteed to detect their respective targets via IHC-P in human samples. As stated in our AbPromise guarantee, we are happy to provide scientific support, replacement or refund should these products if they do not perform as indicated on the datasheet. More information on our Abpromise may be found at the following link:

    https://www.abcam.com/index.html?pageconfig=abpromise

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

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Pancreatic Cancer)
    Specification
    Pancreatic Cancer
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: CC1, a Tris/Borate/EDTA buffer pH 8
    Permeabilization
    No
    Blocking step
    Inhibitor CM as blocking agent for 4 minute(s) · Concentration: 100% · Temperature: 37°C
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    Abcam user community

    Verified customer

    投稿 Dec 18 2012

    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM no band-->even at different concentrations (range from 1:500-1:1000) SAMPLE human Hodgkin-&primary mediastinal B-cell lymphoma cell lines ( L428, L540, HDLM-2, KM-H2, L1236,MedB-1) human tonsil whole protein extract in RIPA lysis buffer PRIMARY ANTIBODY ab8216, anti CD45, mouse monoclonal tried almost every dilution between 1:500 and 1:1000 in milk or only TBST over night incubation at 4?C DETECTION METHOD pierce: super signal west dura POSITIVE AND NEGATIVE CONTROLS USED positive control: human tonsil ANTIBODY STORAGE CONDITIONS 4?C short term storage SAMPLE PREPARATION in RIPA lysis buffer NuPAGE LDS sample buffer heated for 10 Min at 95?C AMOUNT OF PROTEIN LOADED 100?g ELECTROPHORESIS/GEL CONDITIONS 4-12%Bis-Tris gel MOPS running buffer TRANSFER AND BLOCKING CONDITIONS transfer buffer (invitrogen) blocked in 5% milk SECONDARY ANTIBODY pierce - secondary anti mouse antibody (HRP conjugated) dilution: 1:1000 incubation: tried 1h at 37?C to 2,5 hours at RT HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? dilution time of incubation, temperature while incubation buffer blocking ADDITIONAL NOTES I'm a little upset this antibody does not work. CD45 is a real routine antibody for IHC and now it does not work for western blot? Please help me with my problems. I have had enough problems with abcam antibodies such as SHP-1

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    Answer

    Thank you for your enquiry. I am sorry to hear that you have been having difficulties with ab8216. I have read your technical questionnaire and I have a few comments. You are using an extraction protocol that I would advise: RIPA buffer. I also consider your choice of a positive control good in the form of human tonsil. I would not recommend using as much as 100ug in western blotting. However, given that you are obtaining no signal whatsoever, please can you tell me whether you have confirmed the successful transfer of your blot by Ponceau S. Also can you tell me whether you have confirmed the integrity of your lysate preparation using an alternative CD antiserum using these conditions. I look forward to hearing from you.

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    Question

    I bought a cd45 antibody (ab8216) a few weeks ago. Unfortunately it does not work for Western Blot. The datasheet says that it should work for this technique. Could you please help me with that problem because I have already tried different concentrations and treatments, as well as the standard abcam protocol. Please answer me as soon as possible.

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    Answer

    I am sorry to hear that you have been experiencing problems with ab8216 in Western blotting. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 90 days of purchase), and if it appears that the antibody is at fault, a credit note/refund will be offered. Below I enclose a link to a questionnaire which will help you put your protocol information together very easily so we can look into the details of the experiment and provide some help. Thank you, and we look foward to hearing from you. https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=8216&mode=questionaire

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    Question

    What is the epitope recognized by this antibody? Does it recognize any fragments less than the expected size of the protein? I am interested in studying these cleavage fragments.

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    Answer

    We do not know the epitope or if this antibody recognizes any fragments less than the expected size of the protein. We do not have access to the original WB data. We did not perform such a study, and most likely, if we did and saw any smaller fragments, we would consider it as non-specific proteolytic cleavage during sample preparation rather then as a result of some biological process in a cell. Moreover, the major bands migrate between 180-220 kDa, so any cleavage fragment would presumably be lost due to the low density of gel used for LCA isoforms (so the fragments would migrate with the front of the gel). In short, you should test this hypothesis in your particular experimental setup. If you have any additional questions, please contact us again.

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    Question

    Regarding a previous enquiry - could I use the mouse monoclonal antobody on paraffin-embedded mouse tissue in combination with the DAKO animal research kit (for mouse monoclonal antibodies on mouse tissue)? Or is there any anti-CD45 antibody suitable for paraffin-embedded mouse tissue?

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    Answer

    ab8216 has not been tested in mouse, so we have no further information on this. We have several CD45 antibodies so I suggest that you conduct a search through the Abcam Website. I also recommend that you should click on the links “Product Reviews” and “Technical Enquiries” both accessible via the green menu bar on the datasheet. Here you will find customer reviews and past questions and answers specifically relating to that product. If you cannot find what you are looking for within the Abcam catalog, I suggest that you click on the link to "The World's Antibody Gateway". The World's Antibody Gateway is a free search engine service provided by Abcam to help you to quickly find the antibodies that you are looking for. It is a free-text search engine developed by the Abcam team so that it searches the catalogs of all online antibody companies (currently 249).

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    Question

    I am looking for an anti-CD45 antibody that is reactive with sheep tissue (prefaerably paraffin embedded tissues). My questions are: 1. Is ab8216 reactive with sheep tissue or is it likely to be on the basis of between species homology. 2. If not, can you recommend an antibody that does work on sheep tissue. Many thanks

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    Answer

    This antibody has not been tested for cross reactivity with sheep. As noted on the datasheet, for immunohistochemistry, no treatment of parrafin sections neccessary. I suggest that you examine the degree of sequence homology between human and sheep CD45. We do not have an antibody agaisnt CD45 that has already been tested for cross reactivity with sheep, but I suggest that you conduct a search through the Abcam home page, and click on the link in the yellow menu bar to "The World's Antibody Gateway" to determine whether any other companies stock such an antibody. The World's Antibody Gateway is a free search engine service provided by Abcam to help you to quickly find the antibodies that you are looking for. It is a free-text search engine developed by the Abcam team so that it searches the catalogs of all online antibody companies (currently 169).

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    Question

    Hi, May I use this antibody on mouse tissue? Lisa

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    Answer

    No. Even if this antibody did recognise mouse CD45, the antibody has been raised in mouse, therefore the anti mouse secondary would non-specifically bind to the tissue leading to high background staining. At current we do not stock an antibody towards CD45 that has been raised in any other species.

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