Purified Rat Anti-CD11b

BD Pharmingen™ Purified Rat Anti-CD11b

Name: Purified Rat Anti-CD11b
Brand: Purified Rat Anti-CD11b
SKU: 550282

Clone M1/70 (RUO)

Immunohistochemical staining of CD11b positive cells. An acetone-fixed, frozen section of normal mouse spleen was stained with the Purified Rat Anti-CD11b antibody (Cat. No. 550282). Three-step staining was carried out with Biotin Goat Anti-Rat Ig (Cat. No. 559286) as the secondary antibody and Streptavidin HRP (Cat. No. 550946) together with a DAB detection system (Cat. No. 550880). CD11b positive cells can be identified by the intense brown labeling of cell surface membranes (20X magnification).

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Catalog No:550282

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Product Details

Brand: BD Pharmingen™

Alternative Name: Itgam; Integrin alpha-M; Ly-40; Mac-1a; Mac-1 alpha; CR3A; CR-3 alpha chain

Reactivity: Mouse (QC Testing), Human (Tested in Development)

Isotype: Rat DA, also known as DA/HA IgG2b, κ

Immunogen: Mouse Splenic Cells

Application: Flow cytometry (Routinely Tested), Immunohistochemistry-frozen (Tested During Development), Immunohistochemistry-formalin (antigen retrieval required) (Not Recommended)

Concentration: 125 µg/ml

RRID: AB_393577

Storage Buffer: Aqueous buffered solution containing BSA, goat serum, and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Immunohistochemistry: The Purified Rat Anti-Mouse CD11b antibody stains mouse macrophages, granulocytes, dendritic cells, and NK cells. For optimal indirect immunohistochemical staining, investigators are encouraged to titrate this antibody (suggested starting range of a 1:10 to 1:50 dilution) and visualize via a three-step staining procedure using a combination of Biotin Goat Anti-Rat Ig (Cat. No. 559286) as the secondary antibody and Streptavidin HRP (Cat. No. 550946) together with a DAB detection system (Cat. No. 550880). Investigators should note that this antibody is not recommended for formalin-fixed paraffin embedded sections.

Product Notices

Since applications vary, each investigator should titrate the reagent to obtain optimal results.
An isotype control should be used at the same concentration as the antibody of interest.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
This antibody has been developed for the immunohistochemistry application. However, a routine immunohistochemistry test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Data Sheets

Technical Data Sheet (TDS)

Companion Products

Stain Buffer (FBS) RUO

Size 500 ML Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 ML Cat No. 554657

Purified Rat IgG2b, κ Isotype Control RUO

Size 0.25 mg Cat No. 559478

Biotin Goat Anti-Rat Ig RUO

Size 0.5 mg Cat No. 559286

Streptavidin HRP RUO

Size 50 ML Cat No. 550946

DAB Substrate Kit RUO

Size 500 Tests Cat No. 550880

View Details

550282 Rev. 4

Antibody Details

M1/70

The M1/70 monoclonal antibody specifically binds to CD11b, also known as Integrin alpha M (Itgam or αM). CD11b is a 170-kDa type 1 transmembrane glycoprotein and belongs to the Integrin alpha chain family. CD11b serves as the alpha chain of the heterodimeric Mac-1 integrin (CD11b/CD18, αMβ2), also known as complement receptor 3 (CR3). Mac-1 mediates adhesion to ICAM-1 (CD54), ICAM-2 (CD102), fibrinogen and binding to C3bi. Mac-1 is expressed at varying levels on granulocytes, macrophages, myeloid-derived dendritic cells, natural killer cells, microglia, and B-1 B lymphocytes. Mac-1 expression is rapidly upregulated on neutrophils after activation, in the same time period that CD62L (L-selectin) is shed from the cell surface. The M1/70 antibody reportedly blocks cell adherence and C3bi binding but does not block cell-mediated lysis. Cross-reaction of the M1/70 antibody with CD11b expressed on human monocytes, polymorphonuclear leukocytes, and NK cells has been reported.

550282 Rev. 4

Format Details

Purified

Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.

Format: Purified

550282 Rev.4

Citations & References

View Development References

Development References (8)

Ault KA, Springer TA. Cross-reaction of a rat-anti-mouse phagocyte-specific monoclonal antibody (anti-Mac-1) with human monocytes and natural killer cells. J Immunol. 1981; 126(1):359-364. (Biology). View Reference
Flotte TJ, Springer TA, Thorbecke GJ. Dendritic cell and macrophage staining by monoclonal antibodies in tissue sections and epidermal sheets. Am J Pathol. 1983; 111(1):112-124. (Biology). View Reference
Leenen PJ, de Bruijn MF, Voerman JS, Campbell PA, van Ewijk W. Markers of mouse macrophage development detected by monoclonal antibodies. J Immunol Methods. 1994; 174(1-2):5-19. (Biology). View Reference
Sanchez-Madrid F, Simon P, Thompson S, Springer TA. Mapping of antigenic and functional epitopes on the alpha- and beta-subunits of two related mouse glycoproteins involved in cell interactions, LFA-1 and Mac-1. J Exp Med. 1983; 158(2):586-602. (Biology: Blocking, Immunoprecipitation). View Reference
Springer T, Galfre G, Secher DS, Milstein C. Mac-1: a macrophage differentiation antigen identified by monoclonal antibody. Eur J Immunol. 1979; 9(4):301-306. (Biology: Immunoprecipitation). View Reference
Springer T, Galfre G, Secher DS, Milstein C. Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Eur J Immunol. 1978; 8(8):539-551. (Immunogen: Immunoprecipitation). View Reference
Springer TA, Davignon D, Ho MK, Kurzinger K, Martz E, Sanchez-Madrid F. LFA-1 and Lyt-2,3, molecules associated with T lymphocyte-mediated killing; and Mac-1, an LFA-1 homologue associated with complement receptor function. Immunol Rev. 1982; 68:171-195. (Biology: Blocking). View Reference
Vremec D, Zorbas M, Scollay R, et al. The surface phenotype of dendritic cells purified from mouse thymus and spleen: investigation of the CD8 expression by a subpopulation of dendritic cells. J Exp Med. 1992; 176(1):47-58. (Biology). View Reference

View All (8)

550282 Rev. 4

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.