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
Product nameAnti-CD11b antibody [EP1345Y] - C-terminal
See all CD11b primary antibodies
DescriptionRabbit monoclonal [EP1345Y] to CD11b - C-terminal
SpecificityTesting 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.
Tested applicationsSuitable for: ICC/IF, IHC-FoFr, WB, IP, IHC-Pmore details
Species reactivityReacts with: Human
Synthetic peptide within Human CD11b aa 1100 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P11215
- 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
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.
Storage instructionsShipped 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.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|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
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.
For unpurified use at 1 - 5 µg/ml
FunctionIntegrin 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.
Tissue specificityPredominantly expressed in monocytes and granulocytes.
Involvement in diseaseGenetic 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.
Sequence similaritiesBelongs to the integrin alpha chain family.
Contains 7 FG-GAP repeats.
Contains 1 VWFA domain.
DomainThe integrin I-domain (insert) is a VWFA domain. Integrins with I-domains do not undergo protease cleavage.
- Information by UniProt
- antigen CD11b (p170) antibody
- Antigen CD11b p170 antibody
- CD11 antigen like family member B antibody
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
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 128 kDa
Blocking 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
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.
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.
ab52478 has been referenced in 49 publications.
- 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