Western Blot: CD11b/c Antibody [NB110-40766] - Analysis of CD11b/c in Raw 264.7 whole cell lysate.
Immunocytochemistry/ Immunofluorescence: CD11b/c Antibody [NB110-40766] - CD11 antibody was tested in Raw264.7 cells with FITC (green). Nuclei were counterstained with DAPI (blue).
Immunohistochemistry-Frozen: CD11b/c Antibody [NB110-40766] - 10 um Cryosection of murine acute brain slices stained with CD11b/c antibody (1:300 = 3.3 ug/ml) and Alexa Fluor 555 donkey anti rabbit IgG. Antibody shows ...read more
Flow Cytometry: CD11b/c Antibody [NB110-40766] - A surface stain was performed on RAW 264.7 with NB110-40766 and a matched isotype control. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at ...read more
Immunocytochemistry/ Immunofluorescence: CD11b/c Antibody [NB110-40766] - Phase contrast microscopy images of HFA and brain tumor cells and immunofluorescence images of cultured primary brain tumor cells, AA and HFA. ...read more
Immunohistochemistry: CD11b/c Antibody [NB110-40766] - Staining of human bone marrow, myeloid precursors.
In Western blot, a specific band is observed at ~ 160 kDa and an apparent non-specific band is observed at ~ 60 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with citrate buffer (pH 6.0) is recommended. In ICC/IF, membrane staining was observed in Raw 264.7 cells. Use in Immunohistochemistry-Frozen reported in scientific literature (PMID: 29467366).
160 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Rat reactivity reported in scientific literature (PMID: 25150592)
Packaging, Storage & Formulations
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
PBS, 30% Glycerol
0.02% Sodium Azide
Immunogen affinity purified
Alternate Names for CD11b/c Antibody - BSA Free
antigen CD11b (p170)
CD11 antigen-like family member B
CD11Bintegrin, alpha M (complement component receptor 3, alpha; also known as CD11b(p170), macrophage antigen alpha polypeptide)
Cell surface glycoprotein MAC-1 subunit alpha
CR-3 alpha chain
integrin, alpha M (complement component 3 receptor 3 subunit)
Leukocyte adhesion receptor MO1
macrophage antigen alpha polypeptide
neutrophil adherence receptor alpha-M subunit
Neutrophil adherence receptor
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 Mac1 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.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
FAQs for CD11b/c Antibody (NB110-40766). (Showing 1 - 2 of 2 FAQs).
What type of antigen retrieval would you recommend for CD11b/c Antibody (NB110-40766)? I am working with paraformaldehyde-fixed free-floating mouse brain sections (40um).
In regards to your inquiry I have spoken with the lab and while we have tested paraffin embedded sections and not free floating, we would recommend citrate buffer pH 6.0. Since they are free floating the lab indicates you can probably incubate for less time with the buffer. We would also recommend using a no antigen retrieval control. Especially if you wanted to try out a few to see which was the optimal one to use. Please see our product specific IHC-P protocol.
I am trying to stain microglia in paraformaldehyde fixed (overnight), frozen sectioned (25 micron) free floating sections of rat brain with NB110-40766. I have yet to produce any significant staining. I have tried both 5 and 10 ug/ml concentrations of primary antibody and have tried the protocol with and without antigen retrieval using heated sodium citrate. Considering this is a membrane bound protein, could the triton be inhibiting antigen/antibody interaction, and would trying something less harsh such as Tween 20 be worthwhile?
Since our product will detect an antigen in the extracellular domain of the CD11b protein, I think elimination of the Triton X may help substantially. You can try replacing with 0.1% Tween-20, or just eliminating the detergent all together. We have only been able to validate this antibody in paraffin-embedded tissue sections, so we do not have a recommended protocol for frozen or free-floating sections at this time. However, with a fixation in PFA overnight, I think performing some type of antigen retrieval would be highly recommended. From other protocols for antigen retrieval on frozen tissue sections, heat induced with sodium citrate (pH 6.0) is the most commonly used buffer. I think with the modification to the detergent, you should be able to see a signal in your tissue.
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