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RabMAb

Anti-CD11b 抗体 [EP1345Y] (ab52478)

製品の概要

  • 製品名
    Anti-CD11b antibody [EP1345Y]
    CD11b 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP1345Y] to CD11b
  • 特異性
    Testing of mouse and rat tissues (brain, spleen, kidney and heart) in WB gave negative results. However, flow cytometry for mouse RAW 264.7 cell line gave positive results. We have not tested any rat samples in flow cytometry. Due to the variability in mouse, we do not list this as a tested species. We welcome any feedback on mouse and rat reactivity.
  • アプリケーション
    適用あり: ICC/IF, IHC-FoFr, WB, IP, Flow Cyt, IHC-Pmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide corresponding to C terminus of human CD11b

  • ポジティブ・コントロール
    • IHC-P: Human spleen tissue WB: TF1 lysate. THP-1 macrophages, +10 ng/ml LPS. Flow Cytometry: RAW 264.7 cells.
  • 特記事項

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

アプリケーション

Our Abpromise guarantee covers the use of ab52478 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
ICC/IF 1/100 - 1/250.
IHC-FoFr Use at an assay dependent concentration.
WB 1/20000 - 1/50000. Predicted molecular weight: 128 kDa.
IP 1/80.
Flow Cyt 1/20 - 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ターゲット情報

  • 機能
    Integrin alpha-M/beta-2 is implicated in various adhesive interactions of monocytes, macrophages and granulocytes as well as in mediating the uptake of complement-coated particles. It is identical with CR-3, the receptor for the iC3b fragment of the third complement component. It probably recognizes the R-G-D peptide in C3b. Integrin alpha-M/beta-2 is also a receptor for fibrinogen, factor X and ICAM1. It recognizes P1 and P2 peptides of fibrinogen gamma chain.
  • 組織特異性
    Predominantly expressed in monocytes and granulocytes.
  • 関連疾患
    Genetic variations in ITGAM has been associated with susceptibility to systemic lupus erythematosus type 6 (SLEB6) [MIM:609939]. Systemic lupus erythematosus (SLE) is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system.
  • 配列類似性
    Belongs to the integrin alpha chain family.
    Contains 7 FG-GAP repeats.
    Contains 1 VWFA domain.
  • ドメイン
    The integrin I-domain (insert) is a VWFA domain. Integrins with I-domains do not undergo protease cleavage.
  • 細胞内局在
    Membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • antigen CD11b (p170) antibody
    • Antigen CD11b p170 antibody
    • CD11 antigen like family member B antibody
    • CD11 antigen-like family member B antibody
    • CD11b antibody
    • CD11b/CD18 antibody
    • CD49d antibody
    • Cell surface glycoprotein MAC-1 subunit alpha antibody
    • Complement component 3 receptor 3 subunit antibody
    • Complement Component Receptor 3 Alpha antibody
    • Complement receptor type 3, alpha subunit antibody
    • CR 3 alpha chain (CR3A) antibody
    • CR 3 alpha chain antibody
    • CR-3 alpha chain antibody
    • CR3 antibody
    • CR3A antibody
    • F730045J24Rik antibody
    • Integrin Alpha M antibody
    • Integrin alpha M chain antibody
    • Integrin alpha-M antibody
    • Integrin beta 2 alpha subunit antibody
    • Integrin subunit alpha M antibody
    • integrin, alpha M (complement component 3 receptor 3 subunit) antibody
    • ITAM_HUMAN antibody
    • ITGAM antibody
    • Leukocyte adhesion receptor MO1 antibody
    • Ly-40 antibody
    • MAC 1 antibody
    • Mac-1a antibody
    • MAC1 antibody
    • Mac1, alpha subunit antibody
    • MAC1A antibody
    • Macrophage antigen alpha polypeptide antibody
    • MGC117044 antibody
    • Mo1, alpha subunit antibody
    • MO1A antibody
    • Neutrophil adherence receptor alpha M subunit antibody
    • Neutrophil adherence receptor antibody
    • SLEB6 antibody
    see all

Anti-CD11b antibody [EP1345Y] 画像

  • Overlay histogram showing RAW 264.7 cells stained with ab52478 (red line). The cells were fixed with methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52478, 1/50 dilution) for 30 minutes at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1 µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in RAW 264.7 cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • IHC image of CD11b staining in a formalin fixed, paraffin embedded normal human spleen tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab52478, 5 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then 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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • ab52478 staining CD11b in the THP-1 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol. Samples were incubated with primary antibody (1/250). ab150077 was used as the secondary antibody (1/1000). Nuclei were stained with DAPI

  • Immunohistochemical analysis of paraffin-embedded human spleen using ab52478 at a dilution of 1/100.
  • Anti-CD11b antibody [EP1345Y] (ab52478) at 1/20000 dilution + TF1 lysate at 10 µg

    Secondary
    goat anti-rabbit HRP labeled at 1/2000 dilution

    Predicted band size : 128 kDa
    Observed band size : 170 kDa (why is the actual band size different from the predicted?)
  • All lanes : Anti-CD11b antibody [EP1345Y] (ab52478) at 1/5000 dilution

    Lane 1 : THP-1 monocytes, untreated
    Lane 2 : THP-1 monocytes, +10 ng/ml LPS
    Lane 3 : THP-1 macrophages, untreated
    Lane 4 : THP-1 macrophages, +10 ng/ml LPS

    Lysates/proteins at 40 µg per lane.

    Secondary
    IRDye®680-conjugated donkey anti-rabbit IgG
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 128 kDa
    Observed band size : 180 kDa (why is the actual band size different from the predicted?)


    Exposure time : 4 minutes

    This image is courtesy of an anonymous Abreview

    Blocked with 5% milk for 1 hour at 21°C.

    See Abreview

Anti-CD11b antibody [EP1345Y] (ab52478) 使用論文

This product has been referenced in:
  • Bengtsson E  et al. ADAMTS-7 is associated with a high-risk plaque phenotype in human atherosclerosis. Sci Rep 7:3753 (2017). IHC ; Human . Read more (PubMed: 28623250) »
  • Yano K  et al. Cholesterol Efflux Capacity of Apolipoprotein A-I Varies with the Extent of Differentiation and Foam Cell Formation of THP-1 Cells. J Lipids 2016:9891316 (2016). WB ; Human . Read more (PubMed: 27957343) »

See all 26 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (TUBO Breast Cancer Tumor)
Antigen retrieval step
Heat mediated
Permeabilization
Yes - 0.1% Triton X-100
Specification
TUBO Breast Cancer Tumor
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5%
Fixative
Formaldehyde
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投稿 Jun 22 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (THP-1)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
THP-1
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5%
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投稿 Mar 27 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (human cells grown on interphase co-culture)
Permeabilization
No
Specification
human cells grown on interphase co-culture
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Formaldehyde
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投稿 May 24 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
Yes - 3% Triton X 100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

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Verified customer

投稿 Apr 08 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (THP-1 (human monocytic leukemia))
Loading amount
40 µg
Specification
THP-1 (human monocytic leukemia)
Treatment
200 nM PMA for 72 hours (THP-1 macrophages)
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Nov 07 2012

Product Anti-CD11b antibody [EP1345Y] (ab52478) is currently validated for use with human samples types, and is covered by our Abpromise Guarantee only for use with human sample types in the applications listed on the datasheet.

Testing of...

Read More

For Flow Cytometry experiments with ab52478 using RAW 246.7 cells were performed in a 100uL volume, using 1x10^6 cells.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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