I1904-79C

IgG1, Mouse Negative Isotype Control (PE)

Size

£709.00

SKU:: I1904-79C
Application:: Flow Cytometry
Purity:: Purified by Protein G affinity chromatography from tissue culture supernatant.
Storage Conditions:: 4°C Do Not Freeze
Supplier:: United States Biological
Host:: Mouse
Immunogen:: Human T lymphocytes.
Clone:: F8-11-13
Format:: Supplied as a lyophilized powder from PBS, pH 7.2, 5% sucrose, 1% BSA, 0.09% sodium azide. Reconstitute with 1ml sterile ddH2O. Labeled with R-Phycoerythrin (PE).
Extra Details:: Isotype Controls: Isotype controls are a type of negative control designed to measure the level of non-specific background signal caused by primary antibodies, based upon the tissue type of the sample. Non-specific binding of primary antibodies are typically caused by the following reasons: • Non-specific Fc receptor binding to cells • Non-specific antibody interactions with cellular proteins or fluorochromes • Non-specific binding of the antibody or fluorochromes to cellular components • Cell autofluorescence Usually, the background signal is the result of immunoglobulins binding non-specifically to Fc receptors present on the cell surface. For example, antibodies raised in mice, particularly those of the IgG2a isotype, will bind strongly to some human leukocytes regardless of the test antibody specificity. In this case you would need a mouse IgG2a isotype control for use with human cells or tissues. Isotype controls are most commonly used in flow cytometry experiments, but may also be used in immunohistochemistry. Selecting an Isotype Control: Typically, an isotype control is matched to the host species and isotype of your specific primary antibody, the heavy chain (IgA, IgG, IgD, IgE, or IgM), subclass and light chain (kappa, lambda) class of the primary antibody; the same fluorochrome (PE, APC, etc.), and have the same F:P ratio. F:P is a measurement of how many fluorescent molecules are present on each antibody. When using directly labeled primary antibodies, it is also necessary to make sure that the isotype control is conjugated to the same fluorochrome or label as the test antibody. It is not sufficient to use one with a spectrally similar fluorochrome. Non-specific Binding: To avoid non-specific binding, you also need to block Fc receptors on cell types, such as spleen cells, with FcR blocking reagents. Isotype controls should be used at the same concentration/dilution as the primary antibody. Isotype controls are mainly used to determine if the staining is real. Apart from isotype controls, unstained cells should always be included in the experimental set-up to monitor autofluorescence. Applications for Isotype Controls: Although isotype controls are mainly used in flow cytometry, they can be used as standard blocking agents and protein coating agents for other applications including immunofluorescence, immunocytochemistry, western blotting, and ELISA. Applications: Suitable for use in Flow Cytometry used as a matched isotype negative control reagent to asses the level of non-specific binding of mouPurity: Purified by Ion Exchange chromatography. Recommended Dilutions: Flow Cytometry: 10ul of the optimal working dilution to label 10e6 cells in 100ul.. Recommend adjusting the concentration of this reagent to the same as that of the test reagent. Optimal working dilutions to be determined by researcher. Recommended Positive Control: Human leukocytes Recommended Negative Control: Negative on the following rat tissues: peripheral blood leukocytes, thymocytes, splenocytes and macrophages. Fusion Partners: Spleen cells from immunized BALB/c mice were fused with cells of the NSI mouse myeloma cell line. Storage and Stability: Store product at 4°C before and after reconstitution. DO NOT FREEZE! Lyophilized and reconstituted products are stable for 12 months at 4°C after receipt. Reconstitute lyophilized powder with sterile ddH2O prior to use and store remainder as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: PE conjugates are sensitive to light. For maximum recovery of product centrifuge the original vial prior to removing the cap.

Shipping Conditions:: Blue Ice