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Rabbit Recombinant Monoclonal CD11b antibody. C-terminal. Suitable for IHC-P, IP, WB, IHC-FoFr, ICC/IF and reacts with Human samples. Cited in 73 publications.


Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBIHC-FoFrICC/IF
Human
Tested
Tested
Tested
Expected
Tested

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

For unpurified use at 1 - 5 μg/ml

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

1/30

Notes

For unpurified use at 1/80

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

For unpurified use at 1/20000 - 1/50000

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

-

Target data

Function

Integrin ITGAM/ITGB2 is implicated in various adhesive interactions of monocytes, macrophages and granulocytes as well as in mediating the uptake of complement-coated particles and pathogens (PubMed:9558116, PubMed:20008295). 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 ITGAM/ITGB2 is also a receptor for fibrinogen, factor X and ICAM1. It recognizes P1 and P2 peptides of fibrinogen gamma chain. Regulates neutrophil migration (PubMed:28807980). In association with beta subunit ITGB2/CD18, required for CD177-PRTN3-mediated activation of TNF primed neutrophils (PubMed:21193407). May regulate phagocytosis-induced apoptosis in extravasated neutrophils (By similarity). May play a role in mast cell development (By similarity). Required with TYROBP/DAP12 in microglia to control production of microglial superoxide ions which promote the neuronal apoptosis that occurs during brain development (By similarity).

Alternative names

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Rabbit Recombinant Monoclonal CD11b antibody. C-terminal. Suitable for IHC-P, IP, WB, IHC-FoFr, ICC/IF and reacts with Human samples. Cited in 73 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EP1345Y

Purification technique

Affinity purification Protein A

Specificity

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.

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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 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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

Biological function summary

CD11b plays a critical role in various immune responses by mediating phagocytosis chemotaxis and cell migration. It is essential for proper leukocyte adherence to endothelial cells and for the engulfment of pathogens or apoptotic cells. CD11b as part of the Mac-1 complex contributes to the immune surveillance and inflammatory processes. This complex interacts with diverse ligands like ICAM-1 fibrinogen and complement component iC3b facilitating an immunological response.

Activity summary

CD11b also known as Integrin alpha M is a protein with a molecular weight of approximately 170 kDa. This protein functions mechanically as an important adhesion molecule. It forms a complex with CD18 to become part of leukocyte-specific beta2 integrins specifically the Mac-1 (CR3) receptor. CD11b is largely expressed on the surface of various myeloid cells including monocytes macrophages granulocytes and a subset of natural killer cells. Researchers often utilize anti-CD11b antibodies and other markers such as APC or CD11b IHC to study its expression in different contexts.

Pathways

CD11b is instrumental within the integrin signaling pathway and the Leukocyte Transendothelial Migration pathway. Its interaction with ICAM-1 is central to the leukocyte adhesion cascade influencing the movement of immune cells across endothelial barriers. CD11b also associates with proteins such as Src family kinases and PI3K which activate downstream signaling responsible for the reorganization of the cytoskeleton and cell movement during immune responses.

Associated diseases and disorders

CD11b is linked to inflammatory conditions like rheumatoid arthritis and systemic lupus erythematosus. Aberrant expression or function of CD11b can result in improper immune regulation contributing to the pathogenesis of these diseases. Its interaction with complement receptor iC3b plays a part in inflammation and tissue damage observed in these conditions. Consequently CD11b represents a potential target for therapeutic intervention in diseases where dysregulated immune activity is a feature.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478), expandable thumbnail
    Image from Sindhu et al PLoS One. 2015 Jul 22;10(7):e0133494. doi: 10.1371/journal.pone.0133494. eCollection 2015. Fig 5.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478), expandable thumbnail
    Image from Martin et al PLoS One. 2014 Feb 5;9(2):e88226. doi: 10.1371/journal.pone.0088226. eCollection 2014. Fig 6.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)

    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.

  • Immunoprecipitation - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478), expandable thumbnail

    Immunoprecipitation - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)

    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.

    All lanes: Immunoprecipitation - Anti-CD11b antibody [EP1345Y] - C-terminal (AB52478)

    Predicted band size: 127 kDa

  • Western blot - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478), expandable thumbnail

    Western blot - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)

    Blocking and diluting buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-CD11b antibody [EP1345Y] - C-terminal (AB52478) at 0.3 µg/mL

    All lanes: TF-1 (Human Erythroleukemia erythroblast) whole cell lysates at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Predicted band size: 127 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478), expandable thumbnail

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)

    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.

  • Western blot - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478), expandable thumbnail

    Western blot - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478)

    All lanes: Western blot - Anti-CD11b antibody [EP1345Y] - C-terminal (AB52478) at 1/20000 dilution

    All lanes: TF-1 (Human bone marrow erythroleukemia cell line) lysate at 10 µg

    Secondary

    All lanes: goat anti-rabbit HRP labeled at 1/2000 dilution

    Predicted band size: 127 kDa

    Observed band size: 170 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11b antibody [EP1345Y] - C-terminal (ab52478), expandable thumbnail

    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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Product protocols

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