Flow cytometric analysis of total Pre-BCR expression by 697 cells. Cells from the Human 697 Pre-B leukemia cell line cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or with PE Mouse Anti-Human Pre-BCR antibody (Cat. No. 569719/569720; solid line histogram) at 0.125 µg/test. The fluorescence histogram showing cell surface and intracellular Pre-BCR expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact 697 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of Pre-BCR expression on viable Human 697 cells. Cells from the Human 697 Pre-B leukemia cell line were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-Human Pre-BCR antibody (Cat. No. 569719/569720; solid line histogram) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing the expression of Pre-BCR (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) 697 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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Flow cytometric analysis of total Pre-BCR expression by 697 cells. Cells from the Human 697 Pre-B leukemia cell line cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or with PE Mouse Anti-Human Pre-BCR antibody (Cat. No. 569719/569720; solid line histogram) at 0.125 µg/test. The fluorescence histogram showing cell surface and intracellular Pre-BCR expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact 697 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of Pre-BCR expression on viable Human 697 cells. Cells from the Human 697 Pre-B leukemia cell line were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-Human Pre-BCR antibody (Cat. No. 569719/569720; solid line histogram) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing the expression of Pre-BCR (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) 697 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
Pre-B Cell Receptor
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Pre-BCR Complexes Purified from Ltk-Pre-BCR Transfectants
Flow cytometry (Routinely Tested), Intracellular staining (flow cytometry) (Tested During Development)
0.2 mg/ml
613
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
PE Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554680
DAPI Solution RUO
Size
1 mg
Cat No.
564907
Antibody Details
HSL2
The HSL2 monoclonal antibody specifically recognizes the human Pre-B cell Receptor (Pre-BCR) that is comprised of the µ heavy chain (µH chain or µHC) and a surrogate light chain (SL chain or SLC) formed by the noncovalent association of CD179a (VpreB) and CD179b (λ5), also known as either CD179a/CD179b or VpreB/λ5). The HSL2 antibody binds to a unique conformational epitope of the Pre-BCR. The Pre-BCR associates with the signal transducing heterodimer, CD79a (Igα):CD79b (Igβ), to form the Pre-BCR Complex. The Pre-BCR is expressed on Pro-B and early Pre-B cells and plays a critical role in B cell development.
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
Citations & References
View product citations for antibody "569720" on CiteAb
Development References (2)
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Tsuganezawa K, Kiyokawa N, Matsuo Y, et al. Expression profile of pre B-cell receptor components in acute lymphoblastic leukemia and its application to the diagnosis and classification of the disease. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:115-117.
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Tsuganezawa K, Kiyokawa N, Matsuo Y, et al. Flow cytometric diagnosis of the cell lineage and developmental stage of acute lymphoblastic leukemia by novel monoclonal antibodies specific to human pre-B-cell receptor.. Blood. 1998; 92(11):4317-24. (Immunogen). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.