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
Related conjugates and formulations
製品の概要
-
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
-
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.
製品の特性
-
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...
-
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
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
-
Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab52478の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
ICC/IF |
1/100 - 1/250.
|
|
IHC-FoFr | (1) |
Use at an assay dependent concentration.
|
WB | (2) |
1/1000. Predicted molecular weight: 128 kDa.
For unpurified use at 1/20000 - 1/50000 |
IP |
1/30.
For unpurified use at 1/80 |
|
IHC-P | (1) |
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 |
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 |
IP
1/30. For unpurified use at 1/80 |
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
-
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
- Entrez Gene: 3684 Human
- Omim: 120980 Human
- SwissProt: P11215 Human
- Unigene: 172631 Human
-
別名
- antigen CD11b (p170) antibody
- Antigen CD11b p170 antibody
- CD11 antigen like family member B antibody
see all
画像
-
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).
-
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.
-
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.
-
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. -
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).
-
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.
-
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
-
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? -
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.
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (67)
ab52478 は 67 報の論文で使用されています。
- Li Y et al. Single-cell RNA sequencing reveals the multi-cellular ecosystem in different radiological components of pulmonary part-solid nodules. Clin Transl Med 12:e723 (2022). PubMed: 35184398
- Huang S et al. Hepatic TGFβr1 Deficiency Attenuates Lipopolysaccharide/D-Galactosamine-Induced Acute Liver Failure Through Inhibiting GSK3β-Nrf2-Mediated Hepatocyte Apoptosis and Ferroptosis. Cell Mol Gastroenterol Hepatol 13:1649-1672 (2022). PubMed: 35202887
- Huang X et al. The molecular, immune features, and risk score construction of intraductal papillary mucinous neoplasm patients. Front Mol Biosci 9:887887 (2022). PubMed: 36090038
- Kalkar P et al. IFN-β mediates the anti-osteoclastic effect of bisphosphonates and dexamethasone. Front Pharmacol 13:1002550 (2022). PubMed: 36386129
- Almeida AF et al. Controlling Macrophage Polarization to Modulate Inflammatory Cues Using Immune-Switch Nanoparticles. Int J Mol Sci 23:N/A (2022). PubMed: 36499452