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66871
4F2hc/CD98 (D3F9D) XP® Rabbit mAb (PE Conjugate)
Antibody Conjugates
Monoclonal Antibody

4F2hc/CD98 (D3F9D) XP® Rabbit mAb (PE Conjugate) #66871

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Flow Cytometry Image 1 - 4F2hc/CD98 (D3F9D) XP® Rabbit mAb (PE Conjugate)

Flow cytometric analysis of SH-SY5Y cells (blue) and HeLa cells (green) using 4F2hc/CD98 (D3F9D) XP® Rabbit mAb (PE Conjugate) (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (PE Conjugate) #5742 (dashed lines).

To Purchase # 66871S
Product # Size Price
66871S
100 µl  (50 tests) $ 358

Supporting Data

REACTIVITY H
SENSITIVITY Endogenous
MW (kDa)
Source/Isotype Rabbit IgG

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated 4F2hc/CD98 (D3F9D) XP® Rabbit mAb #47213.

Product Usage Information

Application Dilution
Flow Cytometry 1:50

Storage

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Protocol

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Flow Cytometry, Live Cell Protocol for Directly Conjugated Antibodies

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix.
  2. Antibody Dilution Buffer: Purchase ready-to-use Flow Cytometry Antibody Dilution Buffer (#13616), or prepare a 0.5% BSA PBS buffer by dissolving 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.

B. Immunostaining

NOTE: Count cells using a hemocytometer or alternative method.

NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to Immunostaining.

NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.

  1. Aliquot desired number of cells into tubes or wells. (Generally, 5x105 to 1x106 cells per assay.)
  2. Pellet cells by centrifugation and remove supernatant.
  3. Resuspend cells in 100 µl of diluted primary antibody, prepared in Antibody Dilution Buffer at a recommended dilution or as determined via titration.
  4. Incubate for 30 min to 1 hr on ice. Protect from light.
  5. Wash by centrifugation in Antibody Dilution Buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 200-500 µl of Antibody Dilution Buffer and analyze on flow cytometer.

posted June 2017

revised June 2020

Protocol Id: 1504

Specificity / Sensitivity

4F2hc/CD98 (D3F9D) XP® Rabbit mAb (PE Conjugate) recognizes endogenous levels of total 4F2hc/CD98 protein. This antibody is predicted to detect multiple isoforms of 4F2hc/CD98.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val509 of human 4F2hc/CD98 protein.

Background

4F2hc is a transmembrane protein that belongs to the solute carrier family. 4F2hc forms heterodimeric complexes with various amino acid transporters such as LAT1 and LAT2 and regulates uptake of amino acids (1-5). 4F2hc is one of the earliest expressed antigens on the surface of activated human lymphocytes (6), hence it is also named CD98. 4F2hc is expressed in all cell types with the exception of platelets, and is expressed at highest levels in the tubules of the kidney and the gastrointestinal tract (7,8). It is localized at the plasma membrane when associated with LAT1 or LAT2 (9) and at the apical membrane of placenta (10). Research studies have shown that 4F2hc is highly expressed in various tumors including glioma (11), ovarian cancer (12), and astrocytomas (13), and it has been implicated in tumor progression and correlated with poor outcome in patients with pulmonary neuroendocrine tumors (14). 4F2hc is also involved in integrin trafficking through association with β1 and β4 integrins, and regulates keratinocyte adhesion and differentiation (15).

  1. Kanai, Y. et al. (1998) J Biol Chem 273, 23629-32.
  2. Mastroberardino, L. et al. (1998) Nature 395, 288-91.
  3. Pfeiffer, R. et al. (1999) EMBO J 18, 49-57.
  4. Pineda, M. et al. (1999) J Biol Chem 274, 19738-44.
  5. Sato, H. et al. (1999) J Biol Chem 274, 11455-8.
  6. Lumadue, J.A. et al. (1987) Proc Natl Acad Sci USA 84, 9204-8.
  7. Rossier, G. et al. (1999) J Biol Chem 274, 34948-54.
  8. Dave, M.H. et al. (2004) J Physiol 558, 597-610.
  9. Bröer, A. et al. (2001) Biochem J 355, 725-31.
  10. Ritchie, J.W. and Taylor, P.M. (2001) Biochem J 356, 719-25.
  11. Okubo, S. et al. (2010) J Neurooncol 99, 217-25.
  12. Kaji, M. et al. (2010) Int J Gynecol Cancer 20, 329-36.
  13. Nawashiro, H. et al. (2006) Int J Cancer 119, 484-92.
  14. Kaira, K. et al. (2011) Oncol Rep 26, 931-7.
  15. Lemaître, G. et al. (2011) J Dermatol Sci 61, 169-79.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.