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

CAT#: CL046P

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


USD 290.00

2 Weeks*

Size
    • 250 ug

Product Images

Specifications

Product Data
Clone Name RB6-8C5
Applications CT, FC, IHC, WB
Recommended Dilution This antibody is suitable for studies of myeloid differentiation stages and their regulations by cytokines. Applications include Flow Cytometry (1,2,3) complement-mediated depletion (4), Western blot staining (5) and both Frozen and Paraffin Sections (6).
Reactivities Mouse
Host Rat
Isotype IgG2b
Clonality Monoclonal
Specificity This monoclonal antibody reacts with the myeloid differentiation antigen Gr-1. (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 (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 (3).
Formulation PBS
State: Purified
State: Liquid purified IgG fraction
Preservative: 0.02% Sodium Azide
Concentration 1.0 mg/ml
Purification Protein G Chromatography
Gene Name lymphocyte antigen 6 complex, locus G
Background The Gr-1 antigen is primarily a marker of myeloid differentiation. In the bone marrow the level of Gr-1 expression is low on immature myeloblasts and increases as the myeloid cells mature to granulocytes. Gr-1 is also expressed on macrophages and transiently on differentiating monocytes. Expression of Gr-1 on a subpopulation of lymphocytes has also been reported.
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 2x10e7 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test).
4. To each tube, add 0.1-0.2 µg* of CL046P.
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 Goat anti-rat IgG (H+L) .
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:
Mouse Strain: BALB/c
Cell Concentration: 1x10e6 cells per test
Antibody Concentration Used: 0.1 µg/10e6 cells
Isotypic Control: Rat IgG2b

Cell Source-Percentage of cells stained above control:
Thymus: 2.0%
Whole Blood Monocytes: 79.1%
Bone Marrow Macrophages: 87.5%

N.B. Appropriate control samples should always be included in any labelling studies.
* For optimal results in various applications, it is recommended that each investigator determine dilutions appropriate for individual use.

Strain Distribution by Flow Cytometry Analysis:
Cell Concentration: 1x10e6 cells per test
Antibody Concentration Used: 0.1 µg/10e6 cells
Strains Tested: BALB/c, C57BL/6, CBA, C3H/He, AKR
Positive: BALB/c, C57BL/6, CBA, C3H/He, AKR
Negative: none
Reference Data

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*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.