CD11b (ITGAM) Rabbit Polyclonal Antibody

CAT#: TA336565

Rabbit Polyclonal Integrin alpha M/CD11b Antibody

Product Images

CD11b staining in IMhu. Cells were stained with antibodies against the CD11b surface antigen. (A) Panel A shows positive staining for CD11b and (B) panel B shows nuclear DAPI staining. Magnitude 40×.

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  Immunohistochemistry: CD11b Antibody - BSA Free TA336565 - Immunohistochemical analysis of CD11b in human renal cancer with TA336565 using 3,3'-Diaminobenzidine (DAB) with hematoxylin counterstain.

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  Immunocytochemistry/Immunofluorescence: CD11b Antibody - BSA Free TA336565 - CD11b Antibody TA336565 - CD11b antibody TA336565 was tested in Raw264.7 cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with 4',6-diamidino-2-phenylindole (DAPI) (blue) and DyLight 550 (red).

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  Flow (Cell Surface): CD11b Antibody - BSA Free TA336565 - CD11b Antibody TA336565 - A surface stain was performed on Raw264.7 cells with DyLight 550-conjugated [TA336565R] (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 10 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to DyLight 550.

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  Flow (Cell Surface): CD11b Antibody - BSA Free TA336565 - An intracellular stain was performed on THP-1 cells with CD11b TA336565 (blue) and a matched isotype control NBP2-24891 (orange). Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

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  Immunocytochemistry/Immunofluorescence: CD11b Antibody - BSA Free TA336565 - Raw264.7 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with CD11b Antibody conjugated to DyLight 650 (NB110-8947C) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

• Dual RNAscope ISH-IHC: CD11b Antibody - BSA Free TA336565 - CD14 mRNA (red) and CD11b protein (green) were detected in formalin-fixed paraffin-embedded tissue sections of human malignant lymph node. ACD's Integrated Co-Detection Workflow was performed using ACD RNAScope Probe Hs-CD14 and CD11b antibody at 1:200 dilution. Tissue was stained on Leica Bond RX using RNAscope (TM) 2.5 LS Reagent Kit-RED, BOND Polymer Refine Detection (DAB) and Hematoxylin, BOND Polymer Refine Red Detection and Hematoxylin and RNA scope (TM) 2.5 LS Green Accessory Pack. Tissue was counterstained with 50% hematoxylin (blue).

• Immunocytochemistry/Immunofluorescence: CD11b Antibody - BSA Free TA336565 - Raw264.7 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with CD11b Antibody conjugated to Alexa Fluor 647 (TA336565AF647) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

• Dual RNAscope ISH-IHC: CD11b Antibody - BSA Free TA336565 - Formalin-fixed paraffin-embedded tissue sections of human lymph node were probed for CD11b mRNA (ACD RNAScope Probe, catalog #555098; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog #TA336565) at 1:50 dilution with 1 hour incubation at room temperature followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes.

• Immunocytochemistry/Immunofluorescence: CD11b Antibody - BSA Free TA336565 - Raw264.7 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with CD11b Antibody conjugated to Alexa Fluor 488 (TA336565AF488) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

• Immunocytochemistry/Immunofluorescence: CD11b Antibody - BSA Free TA336565 - Raw264.7 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with CD11b Antibody (TA336565) at 1ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

• Western Blot: CD11b Antibody - BSA Free TA336565 - Western Blot detection of CD11b in RAW 264.7 whole cell lysates using TA336565.

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Specifications

Product Data

• Applications: FC, ICC/IF, IHC, Simple Western, WB
• Recommended Dilution: Dual RNAscope ISH-IHC, Immunohistochemistry: 1:400, Simple Western: 1:50, Western Blot: 2 ug/mL, Single Cell Western: 1:10, Flow (Cell Surface): 1:10 - 1:1000, Immunohistochemistry-Paraffin: 1:400, Immunocytochemistry/ Immunofluorescence: 1:200, Immunohistochemistry-Frozen, In-situ Hybridization, Flow Cytometry
• Reactivities: Human, Mouse, Rat
• Host: Rabbit
• Clonality: Polyclonal
• Immunogen: A synthetic peptide made to an internal region (within residues 250-350) of the mouse CD11b protein. [Swiss-Prot# P05555]
• Formulation: PBS, 30% glycerol, 0.1% Sodium Azide. Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Concentration: lot specific
• Purification: Immunogen affinity purified
• Conjugation: Unconjugated
• Storage: Store at -20°C as received.
• Stability: Stable for 12 months from date of receipt.
• Predicted Protein Size: 160 kDa
• Gene Name: integrin subunit alpha M
• Database Link:
  NP_000623
  Entrez Gene 16409 MouseEntrez Gene 25021 RatEntrez Gene 3684 Human
  UniProt ID P11215
• Background: CD11b 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 to CR3, the receptor for the iC3b fragment of the third complement component. It probably recognizes the RGD peptide in C3b. CD11b is also a receptor for fibrinogen, factor X and ICAM1. It recognizes P1 and P2 peptides of fibrinogen gamma chain. The Mac-1 CD11b antigen is present on macrophages, granulocytes, natural killer cells, blood monocytes. CD11b is expressed on 8% spleen cells, 44% bone marrow cells and less than 1% of thymocytes and is commonly used as a microglial marker in nervous tissue.
• Synonyms: CD11B; CR3A; MAC-1; MAC1A; MO1A; SLEB6
• Note: This CD11b antibody is useful for Western blot analysis, Immunocytochemistry, Flow Cytometry (PMID 21422470) and IHC-paraffin embedded sections. In Western blot a specific band is observed ~ 160 kDa and an apparant non-specific band is observed ~ 56 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. In ICC/IF, membrane staining was observed in Raw 264.7 cells. This antibody does not appear to work in human with Western blot. Use in Immunohistochemistry-Frozen reported in scientific literature (PMID 23980916)

Reference Data

• Protein Families: Druggable Genome, ES Cell Differentiation/IPS, Transmembrane
• Protein Pathways: Cell adhesion molecules (CAMs), Hematopoietic cell lineage, Leukocyte transendothelial migration, Regulation of actin cytoskeleton