Overview

  • Product name

  • Description

    Rabbit polyclonal to CD11b
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse
    Predicted to work with: Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse CD11b aa 1-100 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab140320)

  • Positive control

    • This antibody gave a positive signal in Raw264.7 whole cell lysate. It also gave a positive signal in Raw264.7 cell line in IF/ICC.

Properties

Applications

Our Abpromise guarantee covers the use of ab128797 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 170 kDa (predicted molecular weight: 127 kDa).
ICC/IF Use a concentration of 1 µg/ml.
Flow Cyt Use 0.1µg for 106 cells.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

IP Use a concentration of 5 µg/ml.

Target

  • Function

    Integrin alpha-M/beta-2 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 with CR-3, the receptor for the iC3b fragment of the third complement component. It probably recognizes the R-G-D peptide in C3b. Integrin alpha-M/beta-2 is also a receptor for fibrinogen, factor X and ICAM1. It recognizes P1 and P2 peptides of fibrinogen gamma chain.
  • Tissue specificity

    Predominantly expressed in monocytes and granulocytes.
  • Involvement in disease

    Genetic variations in ITGAM has been associated with susceptibility to systemic lupus erythematosus type 6 (SLEB6) [MIM:609939]. Systemic lupus erythematosus (SLE) is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system.
  • Sequence similarities

    Belongs to the integrin alpha chain family.
    Contains 7 FG-GAP repeats.
    Contains 1 VWFA domain.
  • Domain

    The integrin I-domain (insert) is a VWFA domain. Integrins with I-domains do not undergo protease cleavage.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • antigen CD11b (p170) antibody
    • Antigen CD11b p170 antibody
    • CD11 antigen like family member B antibody
    • CD11 antigen-like family member B antibody
    • CD11b antibody
    • CD11b/CD18 antibody
    • CD49d antibody
    • Cell surface glycoprotein MAC-1 subunit alpha antibody
    • Complement component 3 receptor 3 subunit antibody
    • Complement Component Receptor 3 Alpha antibody
    • Complement receptor type 3, alpha subunit antibody
    • CR 3 alpha chain (CR3A) antibody
    • CR 3 alpha chain antibody
    • CR-3 alpha chain antibody
    • CR3 antibody
    • CR3A antibody
    • F730045J24Rik antibody
    • Integrin Alpha M antibody
    • Integrin alpha M chain antibody
    • Integrin alpha-M antibody
    • Integrin beta 2 alpha subunit antibody
    • Integrin subunit alpha M antibody
    • integrin, alpha M (complement component 3 receptor 3 subunit) antibody
    • ITAM_HUMAN antibody
    • ITGAM antibody
    • Leukocyte adhesion receptor MO1 antibody
    • Ly-40 antibody
    • MAC 1 antibody
    • Mac-1a antibody
    • MAC1 antibody
    • Mac1, alpha subunit antibody
    • MAC1A antibody
    • Macrophage antigen alpha polypeptide antibody
    • MGC117044 antibody
    • Mo1, alpha subunit antibody
    • MO1A antibody
    • Neutrophil adherence receptor alpha M subunit antibody
    • Neutrophil adherence receptor antibody
    • SLEB6 antibody
    see all

Images

  • Overlay histogram showing RAW 264.7 cells stained with ab128797 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab128797, 0.1μg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (polyclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • CD11b was immunoprecipitated using 0.5mg RAW 264.7 whole cell extract, 5µg of Rabbit polyclonal to CD11b and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, RAW 264.7 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab128797.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 170kDa, non specific bands - 42, 55 and 65kDa: We are unsure as to the identity of this extra band; CD11b

  • Anti-CD11b antibody (ab128797) at 1 µg/ml + RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 127 kDa
    Observed band size: 170 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 42 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes


    CD11b contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. The predicted molecular weight of CD11b is 127 kDa (SwissProt), however we expect to observe a banding pattern around 170 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
  • ICC/IF image of ab128797 stained Raw246.7 cells. The cells were 4% paraformaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab128797, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:

  • Soni S  et al. ATP redirects cytokine trafficking and promotes novel membrane TNF signaling via microvesicles. FASEB J 33:6442-6455 (2019). Read more (PubMed: 30776316) »
  • Hou L  et al. Integrin CD11b mediates a-synuclein-induced activation of NADPH oxidase through a Rho-dependent pathway. Redox Biol 14:600-608 (2018). Read more (PubMed: 29154191) »
See all 5 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (hepatocytes)
Permeabilization
Yes - 0.1% Triton X-100
Specification
hepatocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 16 2015

Abreviews
Application
Western blot
Sample
Rat Tissue lysate - whole (Spinal cord)
Gel Running Conditions
Reduced Denaturing (4-12% Bis Tris Glicine)
Loading amount
10 µg
Specification
Spinal cord
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 18 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (liver)
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 05 2015

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