Testing Data
(Image: CD11b or OX-42 staining of adult mouse cerebral cortex sections showing microglial cells and their processes. Courtesy of Dr. Felix Eckenstein (University of Vermont).)
anti-Human, Mouse Integrin alpha-M (CD11b, C-1, OX-42 Antigen) Antibody | anti-CD11b antibody
Integrin alpha-M (CD11b, C-1, OX-42 Antigen), Affinity Purified Antibody, Chicken
Reactivity
Human, Mouse
Applications
Immunohistochemistry, Western Blot
Synonyms
Integrin alpha-M (CD11b; C-1; OX-42 Antigen); Affinity Purified Antibody; Chicken; anti-CD11b antibody
Reactivity
Human, Mouse
Form/Format
Affinity Purified; Liquid PBS(pH 7.2; 10 mM; isotonic 0.9%, w/v) with sodium azide (0.02%, w/v).Antibody
Concentration
100 ug/m (varies by lot)
Applicable Applications for anti-CD11b antibody
Immunohistochemistry (IHC), Western Blot (WB)
Application Notes
Immunohistochemistry: 1:25-1:50
Dilutions listed as a recommendation. Optimal dilution should be determined by investigator.
Immunostaining Cell Cultures
1. Draw of culture medium with aspirator and add 1 ml of 3.7 % formalin in PBS solution to the dish. (make up from 10 mls Fisher 37% formalin plus 90mls PBS, the Fisher formalin contains 37% formaldehyde plus about 1 % methanol which may be relevant sometimes). Let sit at room temp for 1 minute. (can add 0.1 % Tween 20 to PBS used here and all subsequent steps to reduce background; probably best not to do this first time round though as it may extract your antigen or help wash your cells off the dish).
2. Take off the formalin/PBS and add 1 ml of cold methanol (-20°C, kept in well-sealed bottle in fridge). Let sit for no more than 1 minute.
3. Take off methanol and add 1 ml of PBS, not letting the specimen dry out. To block nonspecific antibody binding can add ~10ml (=1 %) of goat serum, and can incubate for 30 minutes. Can then add antibody reagents. Typically 100ml of hybridoma tissue culture supernatent or 1 ml of mouse ascites fluid or crude serum. Incubate for 1 hour at room temp. (or can go at 3rC for 30 minutes to 1 hour, or can do 4°C overnight, exact time not too critical). Can do very gentle shaking forwell adherent cell lines (3T3, Hek293 etc.).
4. Remove primary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS.
5. Add 0.5 mls of secondary antibody. These are fluorescently labeled Goat anti mouse or rabbit antibodies and are conjugated to ALEXA dyes and are from Molecular probes (Eugene Oregon, the ALEXA dyes are sulphonated rhodamine compounds and are much more stable to UV than FITC, TRITC, Texas red etc.). Typically make 1:2,000 dilutions of these secondaries in PBS plus 1 % goat serum, BSA or nonfat milk carrier. Incubate for 1 hour at room temp. (or can go at 37 degree C for 30 minutes to 1 hour, or can do 4 degree C overnight). Can do gentle shaking for well adherent cell lines (3T3, HEK293 etc.).
6. Remove secondary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS.
7. Drop on one drop of Fisher mounting medium onto dish and apply 22mm square coverslip. View in the microscope!
Immunostaining Tissue
Solutions:
PBS - sodium phosphate-buffered (100 mM; pH 7.2) isotonic (0.9% NaCl, w/v) saline Antibody dilution buffer (PBS with 0.1% non-ionic detergent, such as Triton X-100 or Tween-20).
Dilutions listed as a recommendation. Optimal dilution should be determined by investigator.
Immunostaining Cell Cultures
1. Draw of culture medium with aspirator and add 1 ml of 3.7 % formalin in PBS solution to the dish. (make up from 10 mls Fisher 37% formalin plus 90mls PBS, the Fisher formalin contains 37% formaldehyde plus about 1 % methanol which may be relevant sometimes). Let sit at room temp for 1 minute. (can add 0.1 % Tween 20 to PBS used here and all subsequent steps to reduce background; probably best not to do this first time round though as it may extract your antigen or help wash your cells off the dish).
2. Take off the formalin/PBS and add 1 ml of cold methanol (-20°C, kept in well-sealed bottle in fridge). Let sit for no more than 1 minute.
3. Take off methanol and add 1 ml of PBS, not letting the specimen dry out. To block nonspecific antibody binding can add ~10ml (=1 %) of goat serum, and can incubate for 30 minutes. Can then add antibody reagents. Typically 100ml of hybridoma tissue culture supernatent or 1 ml of mouse ascites fluid or crude serum. Incubate for 1 hour at room temp. (or can go at 3rC for 30 minutes to 1 hour, or can do 4°C overnight, exact time not too critical). Can do very gentle shaking forwell adherent cell lines (3T3, Hek293 etc.).
4. Remove primary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS.
5. Add 0.5 mls of secondary antibody. These are fluorescently labeled Goat anti mouse or rabbit antibodies and are conjugated to ALEXA dyes and are from Molecular probes (Eugene Oregon, the ALEXA dyes are sulphonated rhodamine compounds and are much more stable to UV than FITC, TRITC, Texas red etc.). Typically make 1:2,000 dilutions of these secondaries in PBS plus 1 % goat serum, BSA or nonfat milk carrier. Incubate for 1 hour at room temp. (or can go at 37 degree C for 30 minutes to 1 hour, or can do 4 degree C overnight). Can do gentle shaking for well adherent cell lines (3T3, HEK293 etc.).
6. Remove secondary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS.
7. Drop on one drop of Fisher mounting medium onto dish and apply 22mm square coverslip. View in the microscope!
Immunostaining Tissue
Solutions:
PBS - sodium phosphate-buffered (100 mM; pH 7.2) isotonic (0.9% NaCl, w/v) saline Antibody dilution buffer (PBS with 0.1% non-ionic detergent, such as Triton X-100 or Tween-20).
Type Info
Chicken IgY
Immunogen
Synthetic peptide-.Keyhole Limpet Hemocyanin (KLH)-conjugate of a synthetic peptide corresponding to a region near the center of the extracellular domain of the CD11b protein.
Other Reagents
Fluorescein-labeled goat anti-chicken IgY1.
1. Prepare your tissue sections or cultured cells as you normally would. Wash your sections or cells for 1 min with PBS at room temperature.
2. Incubate your sections or cells with your chicken primary antibodies (diluted in "antibodydilution buffer") for at least 1 hour at room temperature. The concentration of your antibody maybe anywhere from 1:50-1:150 depending on the titre of the antibody and the concentration ofyour antigen.
3. Wash your sectionsor cells over a 10 minute period at room temperature (with two changes ofPBS).
4. Incubate your sections or cells with fluorescein-labeled goat anti-chicken IgY (1:500 dilution in"antibody dilution buffer” for 1 hour at room temperature. Be sure to keep these slides or culturedishes in subdued light (e.g., in a drawer) to avoid bleaching of the fluorescein dye.
5. Repeat step #4
6. Add a drop of "fluorescence anti-fading reagent" (i-BRITE Plus) to your sections or cells. Placea coverslip over the section. If you want to reduce messiness, you may also seal the coverslipby painting the edges with nail polish.
7. Store the slides or culture dishes in the refrigerator (in the dark).
1. Prepare your tissue sections or cultured cells as you normally would. Wash your sections or cells for 1 min with PBS at room temperature.
2. Incubate your sections or cells with your chicken primary antibodies (diluted in "antibodydilution buffer") for at least 1 hour at room temperature. The concentration of your antibody maybe anywhere from 1:50-1:150 depending on the titre of the antibody and the concentration ofyour antigen.
3. Wash your sectionsor cells over a 10 minute period at room temperature (with two changes ofPBS).
4. Incubate your sections or cells with fluorescein-labeled goat anti-chicken IgY (1:500 dilution in"antibody dilution buffer” for 1 hour at room temperature. Be sure to keep these slides or culturedishes in subdued light (e.g., in a drawer) to avoid bleaching of the fluorescein dye.
5. Repeat step #4
6. Add a drop of "fluorescence anti-fading reagent" (i-BRITE Plus) to your sections or cells. Placea coverslip over the section. If you want to reduce messiness, you may also seal the coverslipby painting the edges with nail polish.
7. Store the slides or culture dishes in the refrigerator (in the dark).
Preparation and Storage
Store at 4°C in the dark. Under these conditions, the antibody should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze the antibody unless you want to store it for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half its binding activity is lost.
Testing Data
(Image: CD11b or OX-42 staining of adult mouse cerebral cortex sections showing microglial cells and their processes. Courtesy of Dr. Felix Eckenstein (University of Vermont).)
Testing Data
(Image: CD11b or OX-42 staining of adult mouse cerebral cortex sections showing microglial cells and their processes. Courtesy of Dr. Felix Eckenstein (University of Vermont).)
Related Product Information for anti-CD11b antibody
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 also a marker for hematopoietic/neural stem and progenitor cells and microglia.
Mutations in the CD11b protein have been assosicated with the autoimmune disease lupus, and CD11b antibodies are therefore commonly in immunology studies.
Mutations in the CD11b protein have been assosicated with the autoimmune disease lupus, and CD11b antibodies are therefore commonly in immunology studies.
Product Categories/Family for anti-CD11b antibody
Similar Products
Product Notes
The CD11b (Catalog #AAA555372) is an Antibody and is intended for research purposes only. The product is available for immediate purchase. The Integrin alpha-M (CD11b, C-1, OX-42 Antigen), Affinity Purified Antibody, Chicken reacts with Human, Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's Integrin alpha-M (CD11b, C-1, OX-42 Antigen) can be used in a range of immunoassay formats including, but not limited to, Immunohistochemistry (IHC), Western Blot (WB). Immunohistochemistry: 1:25-1:50 Dilutions listed as a recommendation. Optimal dilution should be determined by investigator. Immunostaining Cell Cultures 1. Draw of culture medium with aspirator and add 1 ml of 3.7 % formalin in PBS solution to the dish. (make up from 10 mls Fisher 37% formalin plus 90mls PBS, the Fisher formalin contains 37% formaldehyde plus about 1 % methanol which may be relevant sometimes). Let sit at room temp for 1 minute. (can add 0.1 % Tween 20 to PBS used here and all subsequent steps to reduce background; probably best not to do this first time round though as it may extract your antigen or help wash your cells off the dish). 2. Take off the formalin/PBS and add 1 ml of cold methanol (-20°C, kept in well-sealed bottle in fridge). Let sit for no more than 1 minute. 3. Take off methanol and add 1 ml of PBS, not letting the specimen dry out. To block nonspecific antibody binding can add ~10ml (=1 %) of goat serum, and can incubate for 30 minutes. Can then add antibody reagents. Typically 100ml of hybridoma tissue culture supernatent or 1 ml of mouse ascites fluid or crude serum. Incubate for 1 hour at room temp. (or can go at 3rC for 30 minutes to 1 hour, or can do 4°C overnight, exact time not too critical). Can do very gentle shaking forwell adherent cell lines (3T3, Hek293 etc.). 4. Remove primary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS. 5. Add 0.5 mls of secondary antibody. These are fluorescently labeled Goat anti mouse or rabbit antibodies and are conjugated to ALEXA dyes and are from Molecular probes (Eugene Oregon, the ALEXA dyes are sulphonated rhodamine compounds and are much more stable to UV than FITC, TRITC, Texas red etc.). Typically make 1:2,000 dilutions of these secondaries in PBS plus 1 % goat serum, BSA or nonfat milk carrier. Incubate for 1 hour at room temp. (or can go at 37 degree C for 30 minutes to 1 hour, or can do 4 degree C overnight). Can do gentle shaking for well adherent cell lines (3T3, HEK293 etc.). 6. Remove secondary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS. 7. Drop on one drop of Fisher mounting medium onto dish and apply 22mm square coverslip. View in the microscope! Immunostaining Tissue Solutions: PBS - sodium phosphate-buffered (100 mM; pH 7.2) isotonic (0.9% NaCl, w/v) saline Antibody dilution buffer (PBS with 0.1% non-ionic detergent, such as Triton X-100 or Tween-20). . Researchers should empirically determine the suitability of the CD11b for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Integrin alpha-M (CD11b, C-1, OX-42 Antigen), Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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