rev: July 29, 2003

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CD CLUSTERED ANTIBODIES  

(anti-Human and others as indicated)

NOTE: 10% Discount on purchases of  3 or more any combination of CD antibodies. Please inquire for bulk or custom formulations (without preservative or carrier).

RDI Divison of researchd Industries Intl  offers a wide line of  antibodies. Since no one antibody works best for all applications (flow cytometry, neutralization, blotting, histochemistry, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below. All products are for in vitro research use only-not for use in or on humans or animals-not for use in diagnostics. Price/availability/specifications subject to change without notice.


CD11b Antibodies


Mouse anti-human CD11b antibodies

-2 clones     CLB--mon-gran/1(44) (purified & FITC)

                    44  (purified, FITC & PE)


CATALOG#                 CLONE#                  Workshop  Host         Form        Price

RDI-M1386clb              CLB-mon-gran/1(44)       IV       mIgM         purified    $375.00

RDI-M1668clb                " "                                   "          "                FITC       $375.00

RDI-CBL145                 44                                   IV       IgG1           purified    $375.00

RDI-CBL145FT             " "                                   "          "                FITC       $375.00

RDI-CBL145PE             " "                                   "          "                PE           $469.00

see also below


Mouse anti-human CD11b:FITC conjugated

cat# RDI-CD11B-44FT       $375.00/120T

       RDI-CD11B-44BT       $375.00/100ug Biotin conjugated

      RDI-CD11B-44PE        $500.00/120T PE conjugated

      RDI-CD11B-44              $281.00/100ug unconjugated

Presentation: 120 Tests in 0.2ml 50 mM Sodium Phosphate pH 7.5, 100 mM Potassium Chloride, 150mM NaCl. 5% glycerol, 0.2% BSA and 0.04% sodium azide.

Clone: ICRF44

Isotype: Mouse IgG1

Immunogen: Human Monocytes

PRODUCTION: Protein A purified antibody from tissue culture supernatant was reacted with FITC. Unconjugated FITC was separated from antibody/FITC conjugate by desalting column. The antibody/FITC conjugate is at 0.5 mg/ml with a Fluorescein/IgG molar ratio of 7.3.

DILUTION: 1:50 (80ul/test)

INFORMATION: Human CD11b (aM integrin) complexes with CD18 (b2 integrin) to form the complement receptor type 3 (CR3) heterodimer which binds to ICAM-1, fibrinogen, C3b, and coagulation factor X. CD11b expression is restricted to leukocytes, mainly to myeloid and NK cells. Antibody ICRF44 recognizes the integrin alpha M subunit (CD11b) of about 165 kd. Antibody ICRF44 blocks homotypic neutrophil and monocyte (FMLP induced) aggregation.

References: V. Mathotra, et al, (1986) Eur J Immunol 16: 1117-1123. B.L. Myones, et al, (1988) J Clin Invest 82: 640-651. Leukocyte Typing IV (W. Knapp, et al, eds.) Oxford University Press, Oxford, (1989) p. 555-558, 560-562. H.R. Petty & R.F. Todd, (1993) J Leuk Biol 54: 492-494. Leukocyte Typing V (S.F. Schlossman, et al, eds.) Oxford University Press, Oxford, (1995) p. 1588-1590.

STORAGE: Store at 2 - 5oC. Freeze/Thawing is not recommended. Protect from light.

PERFORMANCE: 500K ficoll prepared human peripheral white blood cells were washed and preincubated 5 minutes with 20ul of 250ug/ml human IgG (To block non specific binding) after which they were incubated 45 minutes on ice with 80ul of anti-CD11b/FITC at a 1:50 dilution (10 ug/ml). Cells were washed three times, fixed and analyzed on a BD FACS Calibur. A net 15% sub population of the cells stained positive with a mean shift of 1.06 log10 fluorescent units when compared to a Mouse IgG1/FITC negative control (Catalog#RDI-MIGG1FTX2) at a similar concentration. Binding was 88% blocked when cells were pre incubated 10 minutes with 20ul of 0.5 mg/ml anti-CD11b antibody

For In Vitro research Use Only-not for use in or on humans or animals, not for use in diagnostics

Recommended FITC Isotype Control

cat#RDI-MIGG1FTX2 $188.00/vial 100ug $156.00/vial when ordered with antibody

Bulk and presevative free quotes on request


Product Specification: mouse monoclonal anti-human CD11b

PeliCluster  CD11b


CAT#RDI- M1386clb


Test/vial 200


Clone CLB-mon-gran/1, B2


This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with granulocytes. This antibody has been clustered to CD11b in one of the international Workshop on Human White Cell differentiation Antigens


Isotype Mouse, IgM.


Source Ascites fluid of tumour bearing BALB/c mice.


Purification Ammoniumsulphate precipitation and gel filtration.


Packing Each vial contains 1 ml with approximately 0.2 mg/ml monoclonal antibody and 10 mg BSA in 20 mM TRIS and 150 mM NaCl, pH 8.0.


Preservative Mertiolate (0.001%).

Storage and stability Monoclonal antibodies should be stored in the dark at 2-8°C. The reagent is stable until the expiry date stated on the vial label.


Major reactivity The monoclonal antibody is directed against the CD11b- antigen (MO-1) located on the alpha-M chain of LFA-1 complex (Lymphocyte Function-associated Antigen-1), which is expressed on monocytes, granulocytes and NK-cells. The monoclonal antibody does not react with human platelets.


Molecular mass 165 kDa.


Application The monoclonal antibody can be used to detect LFA-1 deficiency in patients suffering from recurrent bacterial infections. Identification of Large Granular Lymphocytes (LGL). Studies of complement receptor function.


Methods Indirect immunofluorescence staining with analysis by flowcytometry or fluorescence microscopy.

APPLICATION  FOR AN INDIRECT IMMUNOFLUORESCENCE TECHNIQUE

Reagents

- FITC-conjugated goat anti-mouse immunoglobulin antiserum (M1204).

  Remove aggregates by centrifugation at 1000 x g for 10 minutes.

- Buffer: Phosphate Buffered Saline (PBS), containing 0.2% Bovine Serum Albumin (BSA).

- PFA: Para-Formaldehyde 1%, pH 7.2, containing 0.2% BSA.

- Microtiter plates (96 wells, V bottom) or tubes.


Procedure

1 Fixate the cells with PFA 1% and wash them with buffer.

2 Prepare a mononuclear cell suspension with a concentration of 1 x 10 7 cells/ml in buffer.

3 Add 45 µl of cell suspension to the microtiter wells or tubes.

4 Add 5 µl monoclonal antibody to the microtiter wells or tubes and mix gently.

5 Incubate for 30 minutes at 2-8 C.

6 Add 100 µl buffer to the microtiter wells or 2 ml buffer to the tubes and centrifuge at 500 x g for 5 minutes.

7 Aspirate the supernatant from the cell pellet and resuspend the cells.

8 Add 200 µl buffer to the microtiter wells or 2 ml buffer to the tubes and centrifuge at 500 x g for 5 minutes.

9 Aspirate the supernatant from the cell pellet and resuspend the cells.

10 Add 50 µl FITC-conjugated goat anti-mouse immunoglobulin serum, solid phase adsorbed with human immunoglobulins,      diluted 1:80.

11 Incubate for 30 minutes at 2-8 C.

12 Repeat step 6-9.

13 Add 200 µl buffer.

14 The number of positive cells can be determined by flowcytometry analysis or by means of fluorescence microscopy.



NOTE: Care should be taken when drawing blood to avoid activation of platelets.

Platelets activation markers are weakly expressed on platelets prepared from directly fixed normal whole blood.

The expression is maximal on thrombin (1 U/ml) stimulated washed platelets.


FOR IN VITRO RESEARCH USE ONLY


mouse monoclonal anti-human CD11b,conjugated to   Fluorescein


PeliCluster CD11b F


CAT#RDI-M1668clb


Test/vial 100


Clone CLB-mon-gran/1, B2


This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with granulocytes. This antibody has been clustered to CD11b in one of the international Workshop on Human White Cell differentiation Antigens


Isotype Mouse, IgM.


Source Ascites fluid of tumour bearing BALB/c mice.


Purification Ammoniumsulphate precipitation and gel filtration.


Conjugation Conjugated with fluorescein iso thiocyanate isomer 1 (FITC).

Molecular F/P ratio between 5.5 - 9.5.


Packing Each vial contains 1 ml FITC conjugated monoclonal antibody and 10 mg BSA in PBS.


Preservative Mertiolate (0.001%).


Storage and stability Monoclonal antibodies should be stored in the dark at 2-8°C. The reagent is stable until the expiry date stated on the vial label.


Major reactivity The monoclonal antibody is directed against the CD11b- antigen (MO-1) located on the alpha-M chain of LFA-1 complex (Lymphocyte Function-associated Antigen-1), which is expressed on monocytes, granulocytes and NK-cells.

The monoclonal antibody does not react with human platelets.


Molecular mass 165 kDa.


Application The monoclonal antibody can be used to detect LFA-1 deficiency in patients suffering from recurrent bacterial infections. Identification of Large Granular Lymphocytes (LGL). Studies of complement receptor function.


Methods Direc immunofluorescence staining with analysis by flowcytometry or fluorescence microscopy.


APPLICATION  in direct  Immunofluorescence techniques

Method with ficoll purified cells


Reagents and materials

- FITC-conjugated monoclonal antibodies.

- Buffer: Phosphate Buffered Saline (PBS), containing 0.2% Bovine Serum Albumine (BSA)

- PFA: Para-Formaldehyde 1%, pH 7.2,  containing 0.2% BSA

- Microtiter plates (96 wells, V bottom) or tybes.


Procedure

1 Prepare a mononuclear cell suspension with a concentratation of 1 x 10 7 cells/ml.

2 Ad 40 µl of cell suspension to microtiter wells or tubes.

3 Add 10 µl of the antibody to the microtiter wells or tubes and mix gently.

4 Incubate for 30 minutes at 2-8°C.

5 Add 150 µl buffer to the microtiter wells or 2 ml buffer to the tubes and centrifuge at 500 x g for 5 minutes.

6 Aspirate the supernatant from the cell pellet and resuspend the cells.

7 Add 200 µl buffer to the microtiter wells or 2 ml buffer to the tubes and centrifuge at 500 x g for 5 minutes.

8 Aspirate the supernatant from the cell pellet and resuspend the cells.

9 Cytoflow fluorometer analysis:

    Add 200 µl buffer to the microtiter wells and transfer this final cell suspension to appropriate testtubes, or add 200 µl             buffer to the tubes.

10 If analysis within 8 hours is not possible add at no. 9, instead of buffer, 200 µl PFA 1%.


Fig. 1

Fluorescense profile of ficoll purified normal cells

(Scatter gates set on the monocyte fraction)

WHOLE  blood method

Due to sophisticated laser flowcytometry we are now able to distinguish populations of human lymphocytes, monocytes and granulocytes by their characteristics in different light scattering. By gating on the appropriate cell cluster (Fig.2) we can easily determine mononuclear cells, which have been immunofluorescently stained with monoclonal antibodies, and enumerate them.


Reagents and materials

- FITC-conjugated monoclonal antibodies.

- Lysing solution (NH 4Cl, pH 7.2).

- Buffer: PBS, containing 0.2 % BSA.

- PFA: Para-Formaldehyde 1%, pH 7.2, containing 0.2 % BSA.

- Tubes.


Procedure

1 Draw blood into a blood collection tube containing EDTA.

2 Deliver 100 µl (*) of well mixed whole blood to the bottom of the testtube.

3 Add 10 µl of the undiluted antibody to the bottom of the testtube, and mix gently.

4 Incubate for 30 minutes at 2-8°C or in an ice-water bath.

5 Mix the tubes and add 2 ml of lysing solution.

6 Incubate for 3-5 minutes at room temperature.

7 When lysing is completed, centrifuge the tubes at 500 x g for 5 minutes.

8 Aspirate the supernatant from the cell pellet, resuspend the cells in 1 ml buffer.

   Place the tubes in the ice-water bath.

9 Analyse the samples within 8 hours.

If analysis within 8 hours is not possible centrifuge the tubes at 500 x g. Aspirate the supernatant from the cell pellet and resuspend the cells in 1 ml PFA 1%.


Fig. 2:

Scatter patern of a whole normal blood sample

R2 - lymphocytes R3 - monocytes R4 - granulocytes


Note:Use our special negative control to determine background fluorescence produced due to Fc binding capacities by mononuclear cells.

The concentration and F/P ratio of our control has been adjusted to our FITC-conjugated monoclonal antibodies. * This method was developed for blood samples with a normal white count. It may be necessary to adjust the quantity of blood for samples with very high or low white counts


FOR IN VITRO RESEARCH USE ONLY


Mouse ANTI-HUMAN CD11b (C3b RECEPTOR) ANTIGEN


PRODUCT CODE :RDI-CBL145    $375.00/vial of 200ug

          cat#RDI-CBL145FT       $375.00/vial of 100 tests FITC labeled

              #RDI-CBL145PE       $469.00/vial of 100 tests PE labeled


CLONE: 44


ISOTYPE: IgG1


SOURCE: Mouse ascitic fluid


PURIFICATION METHOD: Ion exchange chromatography following ammonium sulphate precipitation.


SPECIFICITY: Integrin heterodimer of MW 165,95kD which comprises the receptor (CR3 am chain, CD11b/ CD18) for the complement component C3i. The CR3 receptor is expressed as a surface molecule by circulating monocytes and granulocytes and also by dendritic reticulum cells in sections of lympoid tissue. Expression is increased on activated granulocytes.

Antigen distribution: Granulocytes > 97%

                               Monocytes > 95%

                               LGL cells 58 ± 17%

                               Peripheral blood lymphocytes 10 ± 5%

                               T-cells (CD3+) < 1%

                               B-cells (CD20+) < 1%

                               Thymocytes < 1%


APPLICATIONS:

* Identification of CR3 molecules on monocytes and granulocytes by indirect immunofluorescence staining.

* Identification of CR3 molecules in frozen sections by immunohistochemical staining

* Patients whose cells lack the CR3 may have immune deficiency symptoms and suffer from recurrent bacterial and fungal infections


REFERENCES: Leucocyte Typing III, Oxford University Press (1987)


PRESENTATION: Each monoclonal is delivered in a sealed vial at a concentration of not less than 200µg/2ml in phosphate buffered saline containing 10mM sodium azide and 1mg/ml of bovine serum albumin. We recommend that each laboratory determine an optimum working titre for use in its particular application.


STORAGE: Purified monoclonal antibodies are generally stable for long periods at 4oC. However repeated warming to room temperature and re-cooling may result in a loss of activity and hence effective working titre. It is recommended that monoclonal antibodies are aliquotted upon receipt and then stored at -20C. After thawing they should then be used as a stock solution for up to 60 days. Dilute working solutions are best prepared on the day of use.


For research use only. Not supplied for use in human diagnostic or therapeutic procedures


For bulk (without BSA & without Preservative) use cat#RDI-CBL145-1XP


RDI Divison of researchd Industries Intl

San Jose, 95123 CA Snell ave 658

USA

or 408-780-0908

EMAIL:margaret@cellular-research.com

Ordering terms

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