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Invitrogen™ TNF alpha Monoclonal Antibody (MP6-XT22), PE-eFluor™ 610, eBioscience™

Description
Description: The MP6-XT22 antibody reacts with mouse tumor necrosis factor-alpha (TNF alpha), a 17 kDa cytokine produced by monocytes, macrophages, neutrophils, NK cells and CD4(+) T cells. TNF alpha has cytolytic activity against a range of tumor cells and is important in immune regulation. TNF alpha forms dimers and trimers and also exists as a 26 kDa membrane-bound form. Applications Reported: This MP6-XT22 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This MP6-XT22 antibody has been tested by intracellular staining and flow cytometric analysis of stimulated mouse splenocytes using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to BestProtocols®: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This can be used at less than or equal to 0.25 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is cap...
Specifications
Specifications
| Antigen | TNF alpha |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | MP6-XT22 |
| Concentration | 0.2 mg/mL |
| Conjugate | PE-eFluor 610 |
| Formulation | PBS with 0.09% sodium azide; pH 7.2 |
| Gene | Tnf |
| Gene Accession No. | P06804 |
| Gene Alias | APC1 protein; Cachectin; C-domain 1; C-domain 2; cTNF; DADB-70P7.1; DIF; ICD1; ICD2; Intracellular domain 1; Intracellular domain 2; N-terminal fragment; NTF; RATTNF; Tnf; TNF alpha; TNF superfamily; TNF α; TNF, macrophage-derived; TNF, monocyte-derived; TNFA; TNF-a; TNFalpha; TNF-alpha; Tnfsf1a; TNFSF2; TNFα; TNLG1F; Tumor necrosis factor; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; tumor necrosis factor alpha (cachetin); tumor necrosis factor alpha precursor; tumor necrosis factor ligand 1F; tumor necrosis factor ligand superfamily member 2; Tumor necrosis factor, membrane form; Tumor necrosis factor, soluble form; tumor necrosis factor-alpha; tumor necrosis factor-alpha precursor; tumor-necrosis factor; tumour necrosis factor |
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Frequently Asked Questions (FAQs)
Our options will depend on the samples you are analyzing.
If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.
For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.
Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).
The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.
The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.
As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.
Safety and Handling
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