Ly6g Rat Monoclonal Antibody [Clone ID: RB6-8C5]

CAT#: CL046B

Ly6g rat monoclonal antibody, clone RB6-8C5, Biotin

Conjugation: Biotin FITC



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CNY 3,531.00


货期*
5周

规格
    • 100 ug

Cited in 1 publication.

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Specifications

Product Data
Clone Name RB6-8C5
Applications FC, IHC, WB
Recommend Dilution Flow Cytometry (Ref.1-3).
Immunohistochemistry on Frozen and Paraffin Embedded Sections (Ref.6).
Western blot (Ref.5).
Complement-mediated depletion.
Reactivity Mouse
Host Rat
Clonality Monoclonal
Immunogen Mouse Granulocytes.
Specificity Anti-Mouse Gr-1 monoclonal antibody reacts with the myeloid differentiation antigen Gr-1. (Ref.1,2). 
This 25-30 kDa cell surface antigen is expressed on myeloid cells but not lymphoid or erythroid cells. The expression of the Gr-1 antigen increases with granulocyte maturation (Ref.3) as shown by the distinct populations of bone-marrow cells this monoclonal antibody labels: negative, low positive and high positive. Expression is transient on cells of monocytic lineage (Ref.3).
Formulation PBS containing 0.02% Sodium Azide as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
Label: Biotin
State: Liquid Ig fraction
Concentration lot specific
Purification Protein G Chromatography
Conjugation Biotin
Storage Condition Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Gene Name lymphocyte antigen 6 complex, locus G
Synonyms Gr-1 Granulocyte marker
Note Protocol: FLOW CYTOMETRY ANALYSIS:

Method:
1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with cell separation medium.
2. Wash 2 times.
3. Resuspend the cells to a concentration of 2x107 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 106 cells, representing 1 test).
4. To each tube, add 0.1-0.2 µg* of CL046B per 106 cells.
5. Vortex the tubes to ensure thorough mixing of antibody and cells.
6. Incubate the tubes for 30 minutes at 4°C.
7. Wash 2 times at 4°C.
8. Add 100 µl of secondary antibody FITC Streptavidin at 1/500 dilution.
9. Incubate tubes at 4°C for 30-60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
10. Wash 2 times at 4°C.
11. Resuspend the cell pellet in 50 µl ice cold media B.
12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.

Media:
A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 µl of 2M sodium azide in 100 mls).
B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 µl of 2M sodium azide in 100 mls).

Results:

Tissue Distribution by Flow Cytometry Analysis:
Mouse Strain: BALB/c
Cell Concentration: 1x106 cells per test
Antibody Concentration Used: 0.1 µg/106 cells
Isotypic Control: Rat IgG2b

Cell: Source Percentage of cells stained above control:
Thymus: 1.5%
Whole Blood: Granulocytes: 90.8%, Monocytes: 91.8%
Bone Marrow: Granulocytes: 94.2%, Monocytes: 95.3%

Cell Source: Bone Marrow Monocytes
Percentage of cells stained above control: 95.3%

Strain Distribution by Flow Cytometry Analysis:

Cell Concentration: 1x106 cells per test.
Antibody Concentration Used: 0.1 µg/106 cells.
Strains Tested: BALB/c, C57BL/6, CBA, C3H/He.
Positive: BALB/c, C57BL/6, CBA, C3H/He.
Negative: none.
Reference Data
*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.

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