Invitrogen

CD11b Monoclonal Antibody (M1/70), Super Bright 780, eBioscience™

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CD11b Antibody in Flow Cytometry (Flow) — BALB/c mouse bone marrow cells were stained with CD45R (B220) Monoclonal Antibody, FITC (Product # 11-0452-82) and 0.06 µg of Rat IgG2b kappa Isotype Control, Super Bright 780 (Product # 78-4031-82) (left) or 0.06 µg of CD11b Monoclonal Antibody, Super Bright 780 (right). Total viable cells were used for analysis.

CD11b Antibody in Immunocytochemistry (ICC/IF) — LRRK2 inhibits the fast recycling of receptors internalized through macropinocytosis A-D Primary mouse bone marrow-derived macrophage cells (BMDM) from adult male WT-mLRRK2 BAC transgenic mice were immunostained with anti-mouse CCR5, or CD11b, or MHC II antibody (shown as green signal) and (A) total Rab10 antibody (red signal), or (B) pT73-Rab10 antibody (red signal) as indicated. Representative photomicrographs (from > 20 images analyzed for each condition from n = 3 biologically independent experiments) are shown. (C) Photomicrographs showing the different cargo with respect to Rab10 tubular structures. (D) EEA1 (shown as green signal) with total Rab10 or pT73-Rab10 (shown as red signals) as indicated. Bounding boxes in panels A-D are magnified in ""High Mag"" panels that show individual vesicles. Scale bars represent 10 and 2 mum for ""High Mag"" images. E Calculated percentage of vesicles positive with tubular structures ( n > 30 vesicles analyzed from cells from n = 3 biologically independent experiments) are shown. Each dot represents one vesicle, with significance assessed by Mann-Whitney test. ** represents P < 0.01. Error bars represent +- SEM. F BMDM cells were treated with 0.004% DMSO or 100 nM MLi2 prior to live cell surface staining with Alexa-594 conjugated rat anti-mouse CD11b antibody on ice for 10 min. Three wells of cells from each condition were fixed and imaged for initial surface CD11b binding (shown in red as indicated). Six wells of cells from each cond

CD11b Antibody in Immunohistochemistry (IHC) — Figure 4 Rb1 mitigates inflammation in Ang II-infused mice. C57BL/6J mice were subcutaneously infused with Ang II (1 mug/kg/min) or saline (sham) for 14 days. Ang II-infused mice received daily treatment of either vehicle (Ang II) or Rb1 at 6.25 mg/kg (Rb1-L), 25 mg/kg (Rb1-M) and 100 mg/kg (Rb1-H). ( A ) Plasma was isolated and analyze by ELISA for the level of IL-1 beta, IL-6 and TNF (n=6-8 per group). ( B ) Immunohistochemistry (IHC) was performed to examine the presence of CD68 + cells (in red) in the heart. DAPI (in blue) was counterstained to help visualize the nuclei. Scale bar, 50 mum. ( C ) The number of CD68 + cells in the heart was quantified (n=5 per group). ( D ) IHC was performed to reveal the presence of CD11b + cells (in red) in the heart. DAPI (in blue) counterstaining was performed to visualize the nuclei. Scale bar, 50 mum. ( E ) The number of CD11b + cells in the heart was quantified (n=5 per group). ( F) . C57BL/6J mice were subcutaneously infused with Ang II (1 mug/kg/min) or saline (sham) for 14 days. Ang II-infused mice received daily treatment of either vehicle (Ang II) or Rb1 at 100 mg/kg (Rb1). The cardiac expression of Il1b, II6 , and Tnf was analyzed by real-time qPCR (n=6-7 per group). 18S rRNA was included as the internal control. Relative fold change was plotted against that from the sham controls. Data were expressed as mean+-S.E.M. * P<0.05, ** P<0.01, *** P<0.001.

CD11b Antibody in Immunohistochemistry, Flow Cytometry (IHC, Flow) — Fig. 3 ZD55-IL-24 inhibits melanoma growth in B16-bearing immunocompetent mouse model mainly through the indirect pathway of inducing systemic antitumor immunity. A , B Changes of the immune-related cytokines in tumors treated with ZD55-IL-24. A Heat-map of the down-regulated immunosuppressive cytokines in Fig. 2 C . B Heat-map of the up-regulated immunostimulatory cytokines in Fig. 2 D . C - J Immunohistochemical analysis of immune cell infiltration in tumors, as well as recruitment and activation in spleens. Tumors and spleens resected from B16-bearing C57BL/6 mice receiving PBS or ZD55-IL-24 treatment indicated in Fig. 1 A were analyzed 2 days after the last injection by immunohistochemical staining. C Shown are representative images of tumor sections immunostained for CD8 (red, left panel), CD11b and Ly-6G (yellow, middle panel), and NK1.1 (red, right panel). D Quantification of the CD8 + cells in C ( n = 11). E Quantification of the CD11b + Ly-6G + Ly-6C low neutrophils in C ( n = 3). f Quantification of the NK1.1 + cells in c ( n = 6). G Quantification of the CD8 + cells in H ( n = 9). H Shown are representative images of spleen sections immunostained for CD8 (red, left panel), CD11b and Ly-6G (yellow, middle panel), and NK1.1 (red, right panel). I Quantification of the CD11b + Ly-6G + Ly-6C low neutrophils in H ( n = 3). j Quantification of NK1.1 + cells in h ( n = 9). Nuclei is counterstained with DAPI (blue). Data are mean +- SD. Scale bars, 300 um. Shown is one of t

CD11b Antibody in Immunohistochemistry (PFA fixed) (IHC (PFA)) — Published figure using CD11b monoclonal antibody (Product # 78-0112-82) in Immunohistochemistry (PFA fixed)

Product Details

Applications

Tested Dilution

Publications

Immunohistochemistry (IHC)

View 7 publications 7 publications

Immunohistochemistry (PFA fixed) (IHC (PFA))

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

View 1 publication 1 publication

Flow Cytometry (Flow)

0.125 µg/test

View 34 publications 34 publications

Product Specifications

Species Reactivity

Mouse

Published species

Mouse

Host/Isotype

Rat / IgG2b, kappa

Recommended Isotype Control

Rat IgG2b kappa Isotype Control (eB149/10H5), Super Bright 780, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

M1/70

Conjugate

Super Bright 780

Excitation/Emission Profile

View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

RRID

AB_2722926

Product Specific Information

Description: The M1/70 monoclonal antibody reacts with mouse CD11b, the 165-170 kDa integrin alphaM. CD11b non-covalently associates with CD18 to form alphaMbeta2 integrin (Mac-1) and binds to CD54 (ICAM-1), C3bi, and fibrinogen. Mac-1 is expressed by macrophages, NK cells, granulocytes, activated lymphocytes and mouse B-1 cells in the peritoneal cavity. M1/70 is also cross-reactive to human CD11b, and can be used for the detection of this antigen on human peripheral blood monocytes, granulocytes, and a subset of NK cells. Through interactions with its ligands, CD11b participates in adhesive cell interactions.

Applications Reported: This M1/70 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This M1/70 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This may be used at less than or equal to 0.125 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.

In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.

Target Information

CD11b (integrin alpha-M, ITGAM, integrin alpha-X, ITGAX) is a 165 kDa adhesion molecule that associates non-covalently with integrin beta-2 (CD18). The CD11b/CD18 heterodimeric complex is also known as integrin alpha-M beta-2, Mac-1, and CR3 (complement receptor 3). CD11b is expressed on the surface of monocytes/macrophages, granulocytes, activated lymphocytes, a subset of NK cells, a subset of dendritic cells, and microglia in the brain. CD11b/CD18 functions as the receptor for ICAM-1 (CD54), ICAM-2 (CD102), ICAM-4 (CD242), CD14, CD50, CD23, heparin, iC3b, fibrinogen, and Factor X -these adhesions are critical for cell-cell and cell-matrix interactions. CD11b is expressed on 8% of spleen cells, 44% of bone marrow cells, and less than 1% of thymocytes, and is commonly used as a microglial marker in nervous tissue. The expression of CD11b increases during monocyte maturation and expression levels vary on tissue macrophages. Further, peritoneal macrophages are reported to express higher levels of CD11b than splenic macrophages. Diseases associated with CD11b dysfunction include systemic lupus erythematosus 6 and ITGAM-related susceptibility to systemic lupus erythematosus.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: CD11 antigen-like family member B; CD11B (p170); cell surface glycoprotein MAC-1 alpha subunit; cell surface glycoprotein MAC-1 subunit alpha; complement component receptor 3 alpha-a; complement receptor type 3; CR-3 alpha chain; integrin alpha-M; leukocyte adhesion receptor MO1; Mac-1 alpha; Mac-1 alpha (Mac1A); macrophage antigen alpha; MGC117044

Gene Aliases: Cd11b; CD11b/CD18; CR3; CR3A; F730045J24Rik; Ly-40; Mac-1; Mac-1a; MAC1

Entrez Gene ID: (Mouse) 16409

Molecular Function: integrin