Anti-CD11b 抗体 [EP1345Y] - C-terminal (ab52478)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1345Y] to CD11b - C-terminal
- Suitable for: ICC/IF, IHC-FoFr, WB, IP, IHC-P
- Reacts with: Human
製品の概要
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製品名
Anti-CD11b antibody [EP1345Y] - C-terminal
CD11b 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1345Y] to CD11b - C-terminal -
由来種
Rabbit -
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, IHC-Pmore details -
種交差性
交差種: Human -
免疫原
Synthetic peptide within Human CD11b aa 1100 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P11215 -
ポジティブ・コントロール
- IHC-P: Human spleen and human cervical cancer tissue WB: TF1 lysate. THP-1 macrophages, +10 ng/ml LPS. ICC/IF: THP-1 cell lysate IP: TF-1 whole cell lysate
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特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP1345Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
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 | 1/1000. Predicted molecular weight: 128 kDa. For unpurified use at 1/20000 - 1/50000 |
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IP | 1/30. For unpurified use at 1/80 |
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IHC-P | 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1 - 5 µg/ml |
ターゲット情報
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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
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参照データベース
- Entrez Gene: 3684 Human
- Omim: 120980 Human
- SwissProt: P11215 Human
- Unigene: 172631 Human
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別名
- antigen CD11b (p170) antibody
- Antigen CD11b p170 antibody
- CD11 antigen like family member B antibody
see all
画像
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical cancer tissue sections labeling CD11b with purified ab52478 at 1:1000 dilution (0.28 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0) ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Hematoxylin was used as a counterstain.
Negative control: PBS instead of the primary antibody (inset).
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Unpurified ab52478 staining CD11b in the THP-1 (Human monocytic leukemia cell line) 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.
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Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478) at 0.3 µg/ml (purified) + TF-1 (Human Erythroleukemia erythroblast) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 128 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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ab52478 (purified) at 1:30 dilution (2 µg) immunoprecipitating CD11b in TF-1 (Human bone marrow erythroleukemia cell line) whole cell lysate.
Lane 1: TF-1 whole cell lysate 10 µg (input).
Lane 2: ab52478 + TF-1 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab52478 in TF-1 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)Sindhu et al PLoS One. 2015 Jul 22;10(7):e0133494. doi: 10.1371/journal.pone.0133494. eCollection 2015. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Enhanced expression of monocytes/ macrophage markers in the obese adipose tissue.
The protein expression (intensity) of monocyte/ macrophage markers was detected by immunohistochemistry (IHC) in the adipose tissue samples from lean, overweight, and obese individuals, 10 each. As shown by representative IHC photomicrographs (100× magnification), expression of (A) CD11b was found to be markedly elevated in overweight and obese adipose tissue samples as compared with lean samples.
Paraffin-embedded sections (4 μm thick) of subcutaneous adipose tissue were deparaffinized in xylene and rehydrated through descending grades of ethanol (100%, 95%, and 75%) to water. Antigen retrieval was performed under pressure cooker boiling for 8 min and cooling for 15 min. After washing in PBS, endogenous peroxidase activity was blocked with 3% H2O2 for 30 min and non-specific antibody binding was clocked with 5% nonfat milk for 1hr and 1% bovine serum albumin (BSA) solution for 1hr. Slides were treated overnight with primary antibodies at room temperature. After washing with PBS (0.5% Tween), slides were incubated for 1hr with secondary antibody conjugated with HRP polymer chain and color was developed using 3,3ʹ-diaminobenzidine chromogen substrate. Specimens were washed in running tap water, lightly counterstained with hematoxylin, dehydrated through ascending grades of ethanol (75%, 95%, and 100%), cleared in xylene, and finally mounted in dibutyl phthalate xylene (DPX).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)Martin et al PLoS One. 2014 Feb 5;9(2):e88226. doi: 10.1371/journal.pone.0088226. eCollection 2014. Fig 6. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Duchenne muscular dystrophy (DMD) muscle was co-stained for Neu5Gc (green), ab52478 (red) and DAPI (blue).
For double immunostaining, sections were first stained overnight at 4°C with anti-Neu5Gc after blocking in 10% (Neu5Gc-free) human serum, after blocking in 5 mg/mL BSA, sections were incubated overnight with both primary antibodies without fixation, washed for one hour and incubated with the appropriate secondary antibodies.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)
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 unpurified 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
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Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478) at 1/20000 dilution (unpurified) + TF-1 (Human bone marrow erythroleukemia cell line) 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? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)
Immunohistochemical analysis of paraffin-embedded human spleen tissue using unpurified ab52478 at a dilution of 1/100.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
参考文献 (51)
ab52478 は 51 報の論文で使用されています。
- Konjevic Sabolek M et al. Communication of CD8+ T cells with mononuclear phagocytes in multiple sclerosis. Ann Clin Transl Neurol 6:1151-1164 (2019). PubMed: 31353869
- Liu W et al. Human Inner Ear Immune Activity: A Super-Resolution Immunohistochemistry Study. Front Neurol 10:728 (2019). PubMed: 31354608
- Ziegler AK et al. The effect of resistance exercise upon age-related systemic and local skeletal muscle inflammation. Exp Gerontol 121:19-32 (2019). PubMed: 30905721
- Henrik Heiland D et al. Tumor-associated reactive astrocytes aid the evolution of immunosuppressive environment in glioblastoma. Nat Commun 10:2541 (2019). PubMed: 31186414
- Li D et al. ß-1,3-Glucan/CR3/SYK pathway-dependent LC3B-II accumulation enhanced the fungicidal activity in human neutrophils. J Microbiol N/A:N/A (2019). PubMed: 30721460