12-0113-42

antibody from Invitrogen Antibodies

Targeting: ITGAM CD11B, CR3A, MAC-1

Provider product page for 12-0113-42

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [22]
Comments [0]
Validations
Flow cytometry [1]
Other assay [8]

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Product number: 12-0113-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD11b (activation epitope) Monoclonal Antibody (CBRM1/5), PE, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The CBRM1/5 monoclonal antibody reacts with an activation-specific epitope of human Mac-1. CBRM1/5 binds a subset of Mac-1 molecules on neutrophils and monocytes after stimulation with chemoattractants or phorbol esters but does not recognize Mac-1 on resting myeloid cells. Through interactions with its ligands, Mac-1 participates in adhesive cell interactions. The epitope recognized by this mAb localizes to the I domain on the alpha chain of Mac-1 very close to the ligand binding site in a region that is widely exposed. CBRM1/5 blocks Mac-1 dependent adhesion to fibrinogen and ICAM-1 and inhibits chemoattractant-stimulated adhesion of eosinophils to the Intercellular Adhesion Molecule-1 (ICAM-1). It should be noted that low level activation may occur during processing of freshly drawn blood. Therefore the CBRM1.5 antibody may exhibit some binding to Mac-1 in these unstimulated samples. However, higher levels of Mac-1 expression are observed in activated samples when compared to unstimulated cells. Applications Reported: The CBRM1/5 antibody has been reported for use in flow cytometric analysis. Applications Tested: This CBRM1/5 antibody has been pre-titrated and tested by flow cytometric analysis of resting and activated normal human peripheral blood cells. This can be used at 5 µL (0.25 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. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Conjugate: Yellow dye
Isotype: IgG
Antibody clone number: CBRM1/5
Vial size: 100 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

ITGAM protein structure - 12-0113-42 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3. NLS-RARalpha inhibits the differentiation of NB4 cells. The differentiation rates of each group were detected using flow cytometry. All-trans retinoic acid was used to induce cell differentiation at a concentration of 1 nM. The differentiation rate of cells in the LV-NLS-RARalpha group decreased significantly. **P
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Effects of Gu-4 on the expression of CD11b and the exposure of CD11b active I-domain. In the presence or absence of Gu-4 (40 nmol/ml), whole blood samples from healthy donors were firstly stimulated with or without LPS (100 ng/ml) for 30 mins, then the samples were co-incubated with saturating amounts of PE conjugated CBRM1/5 or PE labeled anti-CD11b antibody sc-1186 for 10 mins, respectively. After removal of red blood cells, different populations of leukocytes were identified by flow cytometry. A: Expression of CD11b active I-domain. Upon LPS stimulation, CD11b active I-domain on neutrophils was greatly increased, but could be markedly supressed by Gu-4 treatment. B: Expression of CD11b. CD11b expression was significantly elevated in LPS-stimulated neutrophils. Gu-4 showed weak inhibitory effect on such elevation. Data represented the results of triplicate experiments. *: p
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Downregulation of RREB1 enhances differentiation of NB4 and HL-60 cells. (A) The protein levels of CD11b and CEBPbeta in NB4 (A) and HL-60 (D) cells were examined by Western blot. Representative Wright-Giemsa staining images showed the change in cell morphology in NB4 (B) and HL-60 (E) cells. Original magnification: 100x. The CD11b + cells in NB4 (C) and HL-60 (F) cells were analyzed by flow cytometry. * p < 0.05, ** p < 0.01, compared with LV-NC.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 1 Extraction, identification, and purification of myeloid-derived suppressor cell and cultivation of dendritic cell. (A) Cells were extracted from the bone marrow of BALB/c mice and stained by monoclonal antibodies. Under flow cytometry, the expressive rate of Gr-1 + MDSC, CD11b + MDSC, CD11c + MDSC, CD80 + MDSC, F4/80 + MDSC, and MHC-II + MDSC were 70.4%, 3.5%, 4.8%, 1.2%, 0.3%, 2.1% respectively. (B) The expressive rate of Gr-1 + CD11b + MDSC was 22.6%. (C) After MACS by CD11b magnetic bead, purification of Gr-1 + CD11b + MDSC reached 84.6%. (D) Most non-antigen-loaded dendritic cells grew adherently, with different sizes, star or spindle shape, and stretching tubers, but some of the cells seemed to have adopted a half-adherent state with rough surface. The expressive rates of CD11c, CD86, and MHC-II on DCs were 10.9%, 3.8%, and 27.9%, respectively, by flow cytometry. (E) On the 7th day, DCs were stimulated and activated by tumor antigens. DCs in the half-adherent state increased obviously with radial spikes and bigger shape. The expressive rates of CD11c, CD86, and MHC-II were 74.8%, 50.3%, and 49.8%, respectively, by flow cytometry. IgG FITC, a homotypic control antibody, was used to set the gate strategy. Scale bar = 100 mumol/liter. MDSC, myeloid-derived suppressor cell; MACS, magnetic-activated cell sorting; DC, dendritic cell; CD, cluster of differentiation
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 4 Lipoxin-mediated changes to the high-affinity conformation of the CD11b receptor in human neutrophils from patients with atherosclerosis versus healthy controls. Whole blood from healthy controls ( n = 5) or patients with atherosclerosis ( n = 5) was exposed to inflammatory stimulus as indicated, either in the absence or presence of lipoxin A 4 (LXA 4 : 500 nM) or lipoxin B 4 (LXB 4 : 500 nM). Neutrophil expression of the CD11b high-affinity conformation was measured by flow cytometry. (A) Neutrophil expression of the CD11b high-affinity conformation was measured as the cellular mean fluorescence intensity (MFI). The expression was measured in controls (white bars) and patients (gray bars). The cells were untreated (Unstim.) or stimulated with chemotactic peptide N-formyl-Met-Leu-Phe (fMLP, 0.4 muM). Representative MFI histograms for CD11b expression and respective conditions are shown for controls (dashed line) and patients (solid line), where the gates were determined using a negative population (gray shaded peaks). (B) LXA 4 (blue bars) and LXB 4 (green greens)-induced changes to the neutrophil expression of the CD11b high-affinity conformation was calculated as the log 2 fold change relative to respective vehicle-treated condition. The samples were stimulated as indicated. The bar graphs show levels of cellular CD11b MFI. Representative histograms for the expression of CD11b and respective conditions are shown for vehicle (black line), LXA 4 (blue line), and LXB
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Characteristics of BM-MSCs. (a) Representative morphology of BM-MSCs. Scale bar = 500 mu m. (b) Representative flow cytometric characterization of cell surface markers expressed on BM-MSCs. Isotypic controls were represented by the gray filled histograms.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5. FAM3D (family with sequence similarity 3, member D) induces neutrophil adhesion and transmigration and activates Mac-1 (macrophage-1 antigen) in neutrophils. A , Representative photographs and quantification of the adhesion of DiI-labeled neutrophils to human umbilical vein endothelial cells (HUVECs) after coculture in the absence or presence of FAM3D neutralization antibody 6D7 (20 mumol/L) or an equal amount of mouse IgG for 2 hours. HUVEC monolayers were stimulated with 4 ng/mL TNF-alpha (tumor necrosis factor alpha) for 24 hours prior to coculture. n=12. One-way analysis of variance (ANOVA) followed by Tukey's test for multiple comparisons, * P
Conjugate: Yellow dye