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Human Lineage Cocktail 4 (lin 4) (CD2, CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, CD56, CD235a)

BD Pharmingen™ Human Lineage Cocktail 4 (lin 4) (CD2, CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, CD56, CD235a)

(RUO)
Human Lineage Cocktail 4 (lin 4) (CD2, CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, CD56, CD235a)
Lineage cocktail 4 staining on enriched human cord blood: Frozen human cord blood mononuclear cells were thawed then enriched using the Human Lineage Cell Depletion Set (Cat No. 560030).  The enriched human cord blood was then stained with the human lineage cocktail 4, APC CD34 (Cat. No 340667), BD Horizon™ BV421 CD38 (Cat. No 562444), APC-H7 CD45RA (Cat. No. 560674), BD Horizon™ BV605 CD90 (Cat. No. 562685) and the viability dye 7-AAD (Cat. No.559925).  Prospective hematopoietic stem cells (HSCs) were identified by first selecting cells based on the scatter profile (left plot), then the Lineage negative and viable cells (center left plot), then the CD34+CD38- cells (center right plot) and then the CD90+CD45RA- cells (right plot). Flow cytometry was performed on a BD LSRFortessa™ flow cytometry system.
Lineage cocktail 4 staining on enriched human cord blood: Frozen human cord blood mononuclear cells were thawed then enriched using the Human Lineage Cell Depletion Set (Cat No. 560030).  The enriched human cord blood was then stained with the human lineage cocktail 4, APC CD34 (Cat. No 340667), BD Horizon™ BV421 CD38 (Cat. No 562444), APC-H7 CD45RA (Cat. No. 560674), BD Horizon™ BV605 CD90 (Cat. No. 562685) and the viability dye 7-AAD (Cat. No.559925).  Prospective hematopoietic stem cells (HSCs) were identified by first selecting cells based on the scatter profile (left plot), then the Lineage negative and viable cells (center left plot), then the CD34+CD38- cells (center right plot) and then the CD90+CD45RA- cells (right plot). Flow cytometry was performed on a BD LSRFortessa™ flow cytometry system.
Product Details
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BD Pharmingen™
Human (QC Testing)
Flow cytometry (Routinely Tested), Fluorescence activated cell sorting (Tested During Development)
RUO
AB_2869433


Description

The FITC Human Lineage Cocktail 4 has been designed to react with cells from the major hematopoietic lineages, such as T lymphocytes, B lymphocytes, monocytes/macrophages, NK cells, erythrocytes, and granulocytes. This pre-diluted cocktail of twelve FITC-conjugated antibodies is designed to label lineage marker-positive cells for exclusion to facilitate the flow cytometric identification of lineage marker-negative hematopoietic stem cells in human cord blood and bone marrow. Components include clone RPA-2.10, which recognizes CD2; HIT3a, which recognizes CD3; RPA-T4, which recognizes CD4; M-T701, which recognizes CD7 ; HIT8a, which recognizes CD8; HI10a, which recognizes CD10; ICRF44, which recognizes CD11b; M5E2, which recognizes CD14; SJ25-C1, which recognizes CD19; 2H7, which recognizes CD20; B159, which recognizes CD56 and GA-R2 (HIR2), which recognizes CD235a. Additional fluorochrome-labeled reagents may be combined with the FITC Human Lineage Cocktail 4, such as CD34, CD38, CD90 and CD45RA conjugates, to characterize hematopoietic stem cell populations from cord blood or bone marrow.

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 with FITC under optimum conditions, and unreacted FITC was removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562722 Rev. 3
Citations & References
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Development References (4)

  1. Baum CM, Weissman IL, Tsukamoto AS, Buckle AM, Peault B. Isolation of a candidate human hematopoietic stem-cell population. Proc Natl Acad Sci U S A. 1992; 89(7):2804-2808. (Biology). View Reference
  2. Bhatia M, Wang JC, Kapp U, Bonnet D, Dick JE. Purification of primitive human hematopoietic cells capable of repopulating immune-deficient mice. Proc Natl Acad Sci U S A. 1997; 94(10):5320-5325. (Biology). View Reference
  3. Pang WW, Price EA, Sahoo D, et al. Human bone marrow hematopoietic stem cells are increased in frequency and myeloid-biased with age. Proc Natl Acad Sci U S A. 2011; 108(50):20012-20017. (Biology). View Reference
  4. Park CY, Majeti R, Weissman IL. In vivo evaluation of human hematopoiesis through xenotransplantation of purified hematopoietic stem cells from umbilical cord blood. Nat Protoc. 2008; 3(12):1932-1940. (Methodology: Flow cytometry, Fluorescence activated cell sorting). View Reference
562722 Rev. 3

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

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.