CD271 (LNGFR) antibodies
human
Contents
1. Description
1.1 Background information
1.2 Applications
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goat cells
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dog cells
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pig cells
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sheep cells
1.3 Recommended antibody dilution
1.1 Background information
1.4 Reagent requirements
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Antigen: CD271 (LNGFR)
2. General protocol for immunofluorescent staining
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Synonym: NGFR (p75); p75NGFR; p75NTR; TNFRSF16
3. Example of immunofluorescent staining with CD271
(LNGFR) antibodies
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Expression patterns: Clone ME20.4-1.H4 recognizes the CD271
antigen, also known as LNGFR (low-affinity nerve growth
factor receptor) or p75 NTR (neurotrophin receptor). CD271
belongs to the low-affinity neurotrophin receptors and the
tumor necrosis factor receptor superfamily. Initially the human
CD271 (LNGFR) was described to be expressed on cells of the
central and peripheral nervous system, and was suggested to
be involved in the development, survival, and differentiation
of cells.1 CD271 (LNGFR) is expressed on mesenchymal stem/
stromal cells (MSCs)2,3,4, follicular dendritic cells5, mesenchymal
cells involved in mesenchymal-epithelial interactions6,
autonomic and sensory neurons7, oligodendrocytes8, astrocytes9,
schwann cells10,11, and neural crest stem cells12 .
4. References
Warnings
Reagents contain sodium azide. Under acidic conditions
sodium azide yields hydrazoic acid, which is extremely toxic.
Azide compounds should be diluted with running water before
discarding. These precautions are recommended to avoid deposits
in plumbing where explosive conditions may develop.
1.2 Applications
1. Description
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This product is for research use only.
ComponentsMonoclonal CD271 (LNGFR) antibodies,
human conjugated to:
1.3 Recommended antibody dilution
Conjugate
Order no.
1 mL
(100 tests)
Order no.
300 µL
(30 tests)
The recommended antibody dilution for all CD271 (LNGFR)
conjugates is 1:11 for up to 107 cells/100 µL of buffer for labeling of
cells and subsequent analysis by flow cytometry.
FITC
130-091-917
130-098-103
VioBright™ FITC
130-104-847
130-104-893
PE
130-091-885
130-098-111
APC
130-091-884
130-098-112
The recommended antibody dilution for CD271 (LNGFR)-Biotin
is 1:33 for up to 10⁷ cells/100 μL of buffer for labeling of cells
and subsequent analysis by flow cytometry. When working with
formaldehyde-fixed or MicroBead-labeled cells use 1:11 for up to
10⁷ cells/100 μL of buffer.
VioBlue®
130-106-553
130-106-597
The antibody is suited for staining of formaldehyde-fixed cells.
PE-Vio®770
130-099-992
130-100-019
Biotin
130-091-883
130-098-098
1.4 Reagent requirements
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CloneME20.4-1.H4 (isotype: mouse IgG1).
Capacity1 mL: 100 tests or up to 10⁹ total cells
300 μL: 30 tests or up to 3×10⁸ total cells.
Product formatAntibodies are supplied in buffer containing
stabilizer and 0.05% sodium azide.
StorageStore protected from light at 2−8 °C. Do not
freeze. The expiration date is indicated on the
vial label.
Cross-reactivity: The CD271 (LNGFR) antibody has been reported
to react with
140-001-292.06
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Identification and enumeration of CD271 (LNGFR)+ cells by
flow cytometry.
monkey cells
Miltenyi Biotec GmbH
Friedrich-Ebert-Straße 68, 51429 Bergisch Gladbach, Germany
Phone +49 2204 8306-0, Fax +49 2204 85197
[email protected]
www.miltenyibiotec.com
Buffer: Prepare a solution containing phosphate-buffered
saline (PBS), pH 7.2, 0.5% bovine serum albumin (BSA),
and 2 mM EDTA by diluting MACS® BSA Stock Solution
(# 130‑091‑376) 1:20 with autoMACS® Rinsing Solution
(# 130‑091‑222). Keep buffer cold (2−8 °C).
▲▲Note: EDTA can be replaced by other supplements such as anticoagulant
citrate dextrose formula-A (ACD-A) or citrate phosphate dextrose (CPD).
Buffers or media containing Ca 2+ or Mg 2+ are not recommended for use.
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(Optional) FcR Blocking Reagent, human (# 130-059-901) to
avoid Fc receptor–mediated antibody labeling.
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(Optional) Tandem Signal Enhancer, human (# 130-099-888)
to reduce non-specific binding of tandem dye–conjugated
antibodies to human cells, especially to monocytes.
Miltenyi Biotec Inc.
2303 Lindbergh Street, Auburn, CA 95602, USA
Phone 800 FOR MACS, +1 530 888 8871, Fax +1 877 591 1060
[email protected]
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(Optional) Conjugated anti-biotin antibodies, e.g., AntiBiotin-PE (# 130-090-756) as secondary antibody reagent in
combination with CD271 (LNGFR)-Biotin.
4. References
1.
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(Optional) For antibodies for additional staining or for isotype
control, refer to www.miltenyibiotec.com/antibodies.
Thomson, T. M. et al. (1988) Expression of human nerve growth factor receptor
on cells derived from all three germ layers. Exp. Cell Res. 174: 533–539.
2.
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(Optional) Propidium Iodide Solution (# 130‑093‑233) for flow
cytometric exclusion of dead cells without fixation.
Caneva, L. et al. (1995) Immuno-electron microscopy characterization of
human bone marrow stromal cells with anti-NGFR antibodies. Blood Cells
Mol. Dis. 21: 73–85.
3.
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(Optional) Fixation and Dead Cell Discrimination Kit
(# 130‑091‑163) for cell fixation and flow cytometric exclusion
of dead cells.
Quirici, N. et al. (2002) Isolation of bone marrow mesenchymal stem cells by
anti-nerve growth factor receptor antibodies. Exp. Hematol. 30: 783–791.
4.
Jones, E. A. et al. (2002) Isolation and characterization of bone marrow
multipotential mesenchymal progenitor cells. Arthritis Rheum. 46: 3349–
3360.
5.
Pezzati, P. et al. (1992) Expression of nerve growth factor receptor
immunoreactivity on follicular dendritic cells from human mucosa associated
lymphoid tissues. Immunology 76: 485–490.
6.
Huber, L. J. and Chao, M. V. (1995) Mesenchymal and neuronal cell expression
of the p75 neurotrophin receptor gene occur by different mechanisms. Dev.
Biol. 167: 227–238.
7.
Kashiba, H. et al. (1995) Coexpression of trk family members and low-affinity
neurotrophin receptors in rat dorsal root ganglion neurons. Brain Res. Mol.
Brain Res. 30: 158–164.
8.
Casaccia-Bonnefil, P. et al. (1996) Death of oligodendrocytes mediated by the
interaction of nerve growth factor with its receptor p75. Nature 383: 716–719.
1. Determine cell number.
9.
2. Centrifuge cell suspension at 300×g for 10 minutes. Aspirate
supernatant completely.
Rudge, J. S. et al. (1994) Neurotrophic factor receptors and their signal
transduction capabilities in rat astrocytes. Eur. J. Neurosci. 6: 693–705.
10.
DiStefano, P. S. and Johnson, E. M. Jr. (1988) Identification of a truncated form
of the nerve growth factor receptor. Proc. Natl. Acad. Sci. U.S.A. 85: 270–274.
11.
Vroemen, M. and Weidner, N. (2003) Purification of Schwann cells by selection
of p75 low affinity nerve growth factor receptor expressing cells from adult
peripheral nerve. J. Neurosci. Methods 124: 135–143.
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2. General protocol for immunofluorescent
staining
Volumes given below are for up to 107 nucleated cells. When working
with fewer than 107 cells, use the same volumes as indicated. When
working with higher cell numbers, scale up all reagent volumes and
total volumes accordingly (e.g. for 2×10⁷ nucleated cells, use twice
the volume of all indicated reagent volumes and total volumes).
3. Resuspend up to 10⁷ nucleated cells per 100 µL of buffer.
4. Add 10 µL of the CD271 (LNGFR) antibody.
5. Mix well and incubate for 10 minutes in the dark in the
refrigerator (2−8 °C).
▲▲Note: Higher temperatures and/or longer incubation times may lead to nonspecific cell labeling. Working on ice requires increased incubation times.
6. Wash cells by adding 1−2 mL of buffer and centrifuge at 300×g
for 10 minutes. Aspirate supernatant completely.
7. (Optional) If CD271 (LNGFR)-Biotin was used, resuspend the
cell pellet in 100 µL of buffer, add 10 µL of anti-biotin antibody,
and continue as described in steps 5 and 6.
8. Resuspend cell pellet in a suitable amount of buffer for analysis
by flow cytometry or fluorescence microscopy.
3. Example of immunofluorescent staining with
CD271 (LNGFR) antibodies
13.
Rozemuller, H. et al. (2010) Prospective isolation of mesenchymal stem
cells from multiple mammalian species using cross-reacting anti-human
monoclonal antibodies. Stem Cells Dev. 19: 1911–1921.
Refer to www.miltenyibiotec.com for all data sheets and protocols.
Warranty
The products sold hereunder are warranted only to be free from defects in
workmanship and material at the time of delivery to the customer. Miltenyi Biotec
GmbH makes no warranty or representation, either expressed or implied, with
respect to the fitness of a product for a particular purpose. There are no warranties,
expressed or implied, which extend beyond the technical specifications of the
products. Miltenyi Biotec GmbH’s liability is limited to either replacement of the
products or refund of the purchase price. Miltenyi Biotec GmbH is not liable for any
property damage, personal injury or economic loss caused by the product.
autoMACS, MACS, Vio, and VioBlue are registered trademarks and VioBright is a
trademark of Miltenyi Biotec GmbH.
Copyright © 2016 Miltenyi Biotec GmbH. All rights reserved.
Counts
Jurkat cells were transfected with the human LNGFR gene and
mixed with untransfected cells. Mixed cells were stained with
fluorochrome-conjugated CD271 (LNGFR) antibodies conjugated
to PE and then analyzed by flow cytometry.
12. Chalazonitis, A. et al. (1998) Age-dependent differences in the effects of GDNF
and NT-3 on the development of neurons and glia from neural crest-derived
precursors immunoselected from the fetal rat gut: expression of GFRalpha-1 in
vitro and in vivo. Dev. Biol. 204 (2): 385–406.
Anti-LNGFR-PE
140-001-292.06
For more examples please refer to the respective product page at
www.miltenyibiotec.com/antibodies.
Unless otherwise specifically indicated, all Miltenyi Biotec products and services
are for research use only and not for diagnostic or therapeutic use.
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