Surface staining of C57BL/6 splenocytes with Anti-Mouse CD23 PE-Cy7 (cat. 25-0232) and Anti-Mouse CD21/CD35 FITC (cat. 11-0211) followed by intracellular staining using the Foxp3 Staining Buffer Set (cat. 00-5523) and 0.125 µg of Anti-Human/Mouse Pax5 PE. Histogram depicts cell populations based on surface stains comprising the marginal zone cells (CD21highCD23-) (blue histogram) or T1 cells (CD21-CD23-) (green histogram).
Contents: Anti-Human/Mouse Pax5 PE Catalog Number: 12-9918 Concentration: 0.2 mg/ml Formulation: aqueous buffer, 0.09% sodium azide, contains stabilizer if necessary Storage Conditions: Store at 2-8°C. Do not freeze. Light sensitive material. Clone: 1H9 Host/Isotype: Rat IgG2a, k
Product Options
Cat. No.
Size
Price
Add Qty to Cart
12-9918-80
25 ug
12-9918-82
100 ug
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The monoclonal antibody 1H9 recognizes both mouse and human Pax5. Pax5, also known as BSAP (B cell specific activator protein), is a member of the paired box (pax) family of transcription factors. Pax5 is the only member of the pax family of transcription factors that is expressed in hematopoietic cells. During embryogenesis, Pax5 is transiently expressed in the brain of mice and in the mesencephalon and spinal cord of humans. Its expression is upregulated early in B cell development at the time of B cell commitment and is maintained throughout most subsequent stages. Suppression of Pax5 is essential for expression of Blimp-1 and the terminal differentiation of plasma cells. In the spleen, expression of Pax5 is higher in marginal zone B cells (B220+ CD21high CD23low) than in other B cells, especially the transition 1 stage (B220+ CD21- CD23-). In addition to its role in B cell development, Pax5 also affects VH-DJH heavy chain recombination as well as influencing the expression of many other B and non-B cell related proteins. Pax5 expression is correlated with many neoplasms. In diffuse large B cell lymphomas (DLBCL) and non-Hodgkin lymphomas Pax5 is often mutated while in B-cell ALL, expression levels are high. Additionally, translocation with elastin, IGH, ETV6, FOXP1, and EVI3 have been identified.
eBioscience recommends that the Foxp3 Buffers (cat. 00-5521) be used for optimal results when using this antibody for intracellular staining and flow cytometric analysis.
Applications Reported
For research use only, not for diagnostic or therapeutic use. This 1H9 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested
This 1H9 antibody has been tested by intracellulular staining using Foxp3 Buffers (cat. 00-5523) and flow cytometric analysis. 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 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Kallies A, Hasbold J, Fairfax K, Pridans C, Emslie D, McKenzie BS, Lew AM, Corcoran LM, Hodgkin PD, Tarlinton DM, Nutt SL. Initiation of plasma-cell differentiation is independent of the transcription factor Blimp-1. Immunity. 2007 May;26(5):555-66. (1H9, WB, PubMed)
Cobaleda C, Schebesta A, Delogu A, Busslinger M. Pax5: the guardian of B cell identity and function. Nat Immunol. 2007 May;8(5):463-70.
Fuxa M, Busslinger M. Reporter gene insertions reveal a strictly B lymphoid-specific expression pattern of Pax5 in support of its B cell identity function. J Immunol. 2007 Mar 1;178(5):3031-7.
