CD11b/c Antibody - BSA Free

Novus Biologicals | Catalog # NB110-40766

Novus Biologicals
100% Guarantee

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all CD11b/c Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal region (within residues 500-600) of the mouse CD11 (b/c) protein. [Swiss-Prot: P05555]

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 25150592)

Localization

Monocytes and granulocytes.

Marker

Microglia Marker, Macrophage Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

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.

Scientific Data Images for CD11b/c Antibody - BSA Free

Western Blot: CD11b/c AntibodyBSA Free [NB110-40766]

Western Blot: CD11b/c AntibodyBSA Free [NB110-40766]

Western Blot: CD11b/c Antibody - BSA Free [NB110-40766] - Analysis of CD11b/c in Raw 264.7 whole cell lysate.
Immunocytochemistry/ Immunofluorescence: CD11b/c Antibody - BSA Free [NB110-40766]

Immunocytochemistry/ Immunofluorescence: CD11b/c Antibody - BSA Free [NB110-40766]

Immunocytochemistry/Immunofluorescence: CD11b/c Antibody - BSA Free [NB110-40766] - CD11 antibody was tested in Raw264.7 cells with FITC (green). Nuclei were counterstained with DAPI (blue).
Immunohistochemistry-Frozen: CD11b/c Antibody - BSA Free [NB110-40766]

Immunohistochemistry-Frozen: CD11b/c Antibody - BSA Free [NB110-40766]

Immunohistochemistry-Frozen: CD11b/c Antibody - BSA Free [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 specific staining, but the background is quite heavy. Image from verified customer review.
Flow Cytometry: CD11b/c Antibody - BSA Free [NB110-40766]

Flow Cytometry: CD11b/c Antibody - BSA Free [NB110-40766]

Flow Cytometry: CD11b/c Antibody - BSA Free [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 room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, DyLight 550.
CD11b-c Antibody - BSA Free-Western Blot-NB110-40766-img0011.jpg
Immunocytochemistry/ Immunofluorescence: CD11b/c Antibody - BSA Free [NB110-40766]

Immunocytochemistry/ Immunofluorescence: CD11b/c Antibody - BSA Free [NB110-40766]

CD11b-c-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NB110-40766-img0010.jpg
Immunohistochemistry: CD11b/c Antibody - BSA Free [NB110-40766]

Immunohistochemistry: CD11b/c Antibody - BSA Free [NB110-40766]

Immunohistochemistry: CD11b/c Antibody - BSA Free [NB110-40766] - Staining of human bone marrow, myeloid precursors.
CD11b/c Antibody - BSA Free

Western Blot: CD11b/c Antibody - BSA Free [NB110-40766] -

Western Blot: CD11b/c Antibody - BSA Free [NB110-40766] - Effects of oltipraz on the expression of CD11b/c, phosphoinositide 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt), & phosphorylated inhibitor of kappa B alpha (p-IkB alpha ) in the spinal cord of the CCI-injured mice. The relative protein levels of (A) CD11b/c, (B) PI3K, (D) p-Akt, & (E) p-IKB alpha on the ipsilateral side of the spinal cord in the CCI-injured mice treated with oltipraz (OLT) or vehicle are represented. The sham-operated mice (SHAM) treated with vehicle were used as controls. (C) Representative examples of blots for CD11b/c (160 kDa), PI3K (130 kDa), & GAPDH (37 kDa), & (F) for p-Akt (60 kDa), Akt (60 kDa), p-IKB alpha (40 kDa) & IKB alpha (40 kDa). CD11b/c & PI3K are expressed relative to GAPDH levels whereas phosphorylated proteins are expressed relative to their corresponding total proteins. In all panels, * denotes significant differences vs. sham-operated mice treated with vehicle (p < 0.05; one-way ANOVA followed by the SNK test). Results are presented as the mean ± SEM; n = 5 samples per experimental group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31234342), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
CD11b/c Antibody - BSA Free

Western Blot: CD11b/c Antibody - BSA Free [NB110-40766] -

Western Blot: CD11b/c Antibody - BSA Free [NB110-40766] - Effects of oltipraz on the expression of CD11b/c, PI3K, p-Akt, & p-IkB alpha in the prefrontal cortex of the CCI-injured mice. The relative protein levels of (A) CD11b/c, (B) PI3K, (D) p-Akt, & (E) p-IKB alpha in the prefrontal cortex of the CCI-injured mice treated with oltipraz (OLT) or vehicle. The sham-operated mice (SHAM) treated with vehicle were used as controls. Representative examples of blots for (C) CD11b/c (160 kDa), PI3K (130 kDa) & GAPDH (37 kDa), & for (F) p-Akt (60 kDa), Akt (60 kDa), p-IKB alpha (40 kDa) & IKB alpha (40 kDa). CD11b/c & PI3K are expressed relative to GAPDH levels whereas phosphorylated proteins are expressed relative to their corresponding total proteins. In all panels, * denotes significant differences vs. sham-operated mice treated with vehicle (p < 0.05; one-way ANOVA followed by the SNK test). Results are presented as the mean ± SEM; n = 4-–5 samples per experimental group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31234342), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
CD11b/c Antibody - BSA Free

Western Blot: CD11b/c Antibody - BSA Free [NB110-40766] -

Western Blot: CD11b/c Antibody - BSA Free [NB110-40766] - Effects of oltipraz on the expression of CD11b/c, PI3K, p-Akt, & p-IkB alpha in the hippocampus of the CCI-injured mice. The relative protein levels of (A) CD11b/c, (B) PI3K, (D) p-Akt, & (E) p-IKB alpha in the hippocampus of the CCI-injured mice treated with oltipraz (OLT) or vehicle. The sham-operated mice (SHAM) treated with vehicle were used as controls. Representative examples of blots for (C) CD11b/c (160 kDa), PI3K (130 kDa), & GAPDH (37 kDa); & (F) for p-Akt (60 kDa), Akt (60 kDa), p-IKB alpha (40 kDa), & IKB alpha (40 kDa). CD11b/c & PI3K are expressed relative to GAPDH levels whereas phosphorylated proteins are expressed relative to their corresponding total proteins. In all panels, * denotes significant differences vs. sham-operated mice treated with vehicle (p < 0.05; one-way ANOVA followed by the SNK test). Results are presented as the mean ± SEM; n = 5 samples per experimental group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31234342), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
CD11b/c Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: CD11b/c Antibody - BSA Free [NB110-40766] -

Immunocytochemistry/ Immunofluorescence: CD11b/c Antibody - BSA Free [NB110-40766] - Phase contrast microscopy images of HFA & brain tumor cells & immunofluorescence images of cultured primary brain tumor cells, AA & HFA.HFA were derived from three different foetal brains. (A) 18-week foetus (10× magnification); (B) 19-week foetus, first week in culture (10× magnification); (C) 19-week foetus, day 5 in culture (20× magnification); (D) Primary GBM after 2 trypsinisations (20x); (E) Recurrent GBM (20x); (F) Secondary GBM after 3 trypsinisations (10x); (G) OII 14 days in culture (10x); (H) AII 13 days in culture (10x); (I) Negative control for NGS merged with DAPI; (J) Negative control for secondary antibodies merged with DAPI; (K) GBM cells derived from one patient – GFAP (DAKO) merged with DAPI & CD68 (zoomed: 2.4× magnification); (L) primary brain tumor cells from one patient (male, 40 years old) (passaged three times) - GFAP (Novocastra) merged with DAPI; (K & L) GFAP (green) was used as a GBM marker & CD68 (red) was used as a marker for microglia. (M) AA from one Female, 61 years old; (passaged twice) - GFAP (Novocastra) merged with DAPI; (N) AA from one male 40 years old; (passaged once) - GFAP (Sigma) merged with DAPI & CD68 (Abcam); (M & N) GFAP (green) was used as an astrocyte marker & CD68 (red) & CD11b (green) were used as markers for microglia; (O) HFA from one 17-week- old foetus (passaged once) - GFAP (Novocastra) merged with DAPI & CD11b (Novus); (P) HFA from an 18-week- old foetus (passaged once) - GFAP (DAKO) merged with DAPI & CD68 (zoomed: 2.1× magnification); (O & P) GFAP (green & red) was used as an astrocyte marker & CD68 (red) & CD11b (green) were used as markers for microglia. Nuclei indicated by DAPI (blue) in all images. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0112945), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for CD11b/c Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

1:50 - 1:200

Immunocytochemistry/ Immunofluorescence

1:50-1:200

Immunohistochemistry

5-10 ug/ml

Immunohistochemistry-Frozen

reported in scientific literature (PMID 29467366)

Immunohistochemistry-Paraffin

5-10 ug/ml

Western Blot

2-4 ug/ml
Application Notes
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.

Reviewed Applications

Read 3 reviews rated 3.7 using NB110-40766 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS + 30% Glycerol

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: CD11b/c

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.

Alternate Names

antigen CD11b (p170), CD11 antigen-like family member B, CD11b, CD11b antigen, 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, CR3AMGC117044, integrin alpha-M, integrin, alpha M (complement component 3 receptor 3 subunit), Leukocyte adhesion receptor MO1, MAC-1, MAC1A, macrophage antigen alpha polypeptide, MO1A, Neutrophil adherence receptor, neutrophil adherence receptor alpha-M subunit, SLEB6

Gene Symbol

ITGAM

Additional CD11b/c Products

Product Documents for CD11b/c Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for CD11b/c Antibody - BSA Free

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.

Citations for CD11b/c Antibody - BSA Free

Customer Reviews for CD11b/c Antibody - BSA Free (3)

3.7 out of 5
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • CD11b/c Antibody
    Name: Kirsten Hattermann-Koch
    Application: Immunohistochemistry-Frozen
    Sample Tested: Mouse brain
    Species: Mouse
    Verified Customer | Posted 03/11/2019
    10 µm Cryosection of murina acute brain slices stained with CD11b/c antibody (1:300 = 3.3 µg/ml) and Alexa Fluor 555 donkey anti rabbit IgG. Antibody shows specific staining, but the background is quite heavy.
    CD11b/c Antibody - BSA Free NB110-40766
  • Name: Carmen Wong
    Application: Immunohistochemistry
    Sample Tested: Tumor sections
    Species: Human
    Verified Customer | Posted 08/04/2010
  • Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Human. Mucous membrane of the small intestine
    Species: Human
    Verified Customer | Posted 01/30/2009

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Protocols

View specific protocols for CD11b/c Antibody - BSA Free (NB110-40766):


Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.


Immunohistochemistry
1. Prepare tissue with formalin fixation and by embedding it in paraffin wax.
2. Make 4-mm sections and place on pre-cleaned and charged microscope slides.
3. Heat in a tissue-drying oven for 45 minutes @ 60 degrees Celcius.
4. Deparaffinize the tissues by wash drying the slides in 3 changes of xylene for 5 minutes each @ RT.
5. Rehydrate the tissues by washing the slides in 3 changes of 100% alcohol for 3 minutes each @ RT.
6. Wash the slides in 2 changes of 95% alcohol for 3 minutes each @ RT.
7. Wash the slides in 1 change of 80% alcohol for 3 minutes @ RT.
8. Rinse the slides in gentle running distilled water for 5 minutes @ RT.
9. Perform antigen retrieval by steaming the slides in 0.01M sodium citrate buffer (pH 6.0) @ 99-100 degrees Celcius for 20 minutes.
10. Remove the slides from the heat and let stand in buffer @ RT for 20 minutes.
11. Rinse the slides in 1X TBS-T for 1 minute @ RT.
**Do not allow the tissues to dry at any time during the staining procedure**
12. Begin the immunostaining by applying a universal protein block for 20 minutes @ RT.
13. Drain protein block from the slides and apply the diluted primary antibody for 45 minutes @ RT.
14. Rinse the slide in 1X TBS-T for 1 minute @ RT.
15. Apply a biotinylated anti-rabbit IgG (H+L) secondary for 30 minutes @ RT.
16. Rinse the slide in 1X TBS-T for 1 minute @ RT.
17. Apply an alkaline phosphatase steptavidin for 30 minutes @ RT.
18. Rinse the slide in 1X TBS-T for 1 minute @ RT.
19. Apply an alkaline phosphatase chromagen substrate for 30 minutes @ RT.
20. Rinse the slide in distilled water for 1 minute @ RT.
**This method should only be used if the chromagen substrate is alcohol insoluble (ie: Vector Red, DAB)**
21. Dehydrate the tissue by washing the slides in 2 changes of 80% alcohol for 1 minute each @ RT.
22. Wash the slides in 2 changes of 95% alcohol for 1 minute each @ RT.
23. Wash the slides in 3 changes of 100% alcohol for 1 minute each @ RT.
24. Wash the slides in 3 changes of xylene for 1 minute each @ RT.
25. Apply cover slip.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

**Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for CD11b/c Antibody - BSA Free

Showing  1 - 2 of 2 FAQs Showing All

  • Q: 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?

    A: 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.

  • Q: 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).

    A: 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.

  • Q: 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?

    A: 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.

  • Q: 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).

    A: 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.

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