Anti-Human CD18/CR3 – Dylight® 550

Anti-Human CD18/CR3 – Dylight® 550

Product No.: C569

[product_table name="All Top" skus="C1543"]

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Clone
KIM127
Formats AvailableView All
Product Type
Monoclonal Antibody
Isotype
IgG1 κ
Applications
ELISA
,
FC
,
IF
,
IF Microscopy
,
IP
,
WB

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Antibody Details

Product Details

Reactive Species
Human
Host Species
Mouse
Product Concentration
0.2 mg/ml
Formulation
This DyLight® 550 conjugate is formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.4, 1% BSA and 0.09% sodium azide as a preservative.
Storage and Handling
This DyLight® 550 conjugate is stable when stored at 2-8°C. Do not freeze.
Country of Origin
USA
Shipping
Next Day Ambient
Excitation Laser
Yellow Laser (563 nm)
Applications and Recommended Usage?
Quality Tested by Leinco
WB3,
FC3,5,7,8,9,10,11,12
Additional Applications Reported In Literature ?
ELISA3,
IP13,
IF Microscopy12, 14, 15, 16
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Specificity
Anti-CD18/CR3 antibody (clone KIM127) recognizes an epitope on the leukocytic cell-adhesion molecule (Leu-CAM) common β chain, CD18. As such, this antigen is considered part of the Leu-CAM β integrin family of adhesion molecules. Activity is directed against immunoprecipitated human CR3 and lymphocytes expressing LFA-1. KIM127 binds to pre-activated integrins.
Background
Integrins are a large family of α/β heterodimeric transmembrane molecules that mediate adhesion, migration, cell survival, and cell differentiation1. The leukocytic cell-adhesion molecule (Leu-CAM) family has three members, LFA-1 (CD18/CD11a), CR3 (CD18/CD11b), and p150,95 (CD18/CD11c), all of which share the β chain CD182.

KIM127 recognizes a temperature sensitive epitope on CD18, binding to lymphocytes that express LFA-1 as well as to immunopurified CR33. KIM127 promotes LFA-1 and CR3-dependent adhesion events and enhances aggregation of the human B lymphoblastoid line JY and human T cell line MOLT-4. This aggregation is completely blocked by anti-LFA-1 antibody DA36. KIM127 is not associated with increased surface expression of CR3 or LFA-1.

KIM127 recognizes residues Gly 504, Leu 506, and Tyr 508 of CD18 in a cysteine-rich repeat located in the long loop protruding from the C-terminal end of the integrin-epidermal growth factor-like (I-EGF) 2 domain4,5,6. The epitope is near the extreme bend (genu) between the integrin headpiece and stalk, buried deep in the crevice formed by the bend between the α subunit thigh and calf-1 domains, and is only accessible when the integrin is extended in its activated conformation6. In resting integrins, the KIM127 epitope is shielded by the ⍺ subunit5. KIM127 preferentially reacts with activated β2 integrins and free β2 subunit compared with resting β2 integrins. A switchblade-like action that extends the headpiece of the integrin away from the plasma membrane is thought to expose the KIM127 epitope when activated6.

Activation of LFA-1 ligand binding by Mn2+, and to a lesser degree by Mg2+ in the absence of Ca2+, is positively correlated with increased expression of the KIM127 epitope5. PMA also induces a small but consistent increase in KIM127 expression.

KIM127 was obtained from a BALB/c mouse immunized with CR3 immunopurified from a human white cell lysate3. Hybridoma lines were created by fusing spleen cells with Sp2/0 cells and screening by ELISA for their ability to bind to immunopurified CR3 and then for their ability to promote aggregation of different cell types. Mildly denaturing conditions are required during Western blotting because boiling the sample destroys the KIM127 epitope.
Antigen Distribution
Members of the Leu-CAM β integrin family of adhesion molecules are widely distributed on circulating leukocytes.

Antigen Details

Research Area
Cell Adhesion

References & Citations

1. Luo BH, Carman CV, Springer TA. Annu Rev Immunol. 25:619-647. 2007.
2. Sanchez-Madrid F, Nagy JA, Robbins E, et al. J Exp Med. 158(6):1785-1803. 1983.
3. Robinson MK, Andrew D, Rosen H, et al. J Immunol. 148(4):1080-1085. 1992.
4. Stephens P, Romer JT, Spitali M, et al. Cell Adhes Commun. 3(5):375-384. 1995.
5. Lu C, Ferzly M, Takagi J, et al. J Immunol. 166(9):5629-5637. 2001.
6. Beglova N, Blacklow SC, Takagi J, et al. Nat Struct Biol. 9(4):282-287. 2002.
7. Xie C, Shimaoka M, Xiao T, et al. Proc Natl Acad Sci U S A. 101(43):15422-15427. 2004.
8. Kuwano Y, Spelten O, Zhang H, et al. Blood. 116(4):617-24. 2010.
9. Biber G, Ben-Shmuel A, Noy E, et al. Nat Commun. 12(1):5581. 2021.
10. Chen J, Yang W, Kim M, et al. Proc Natl Acad Sci U S A. 103(35):13062-7. 2006.
11. Boras M, Volmering S, Bokemeyer A, et al. J Exp Med. 214(3):851-874. 2017.
12. Faridi MH, Altintas MM, Gomez C, et al. Biochim Biophys Acta. 1830(6):3696-710. 2013.
13. Jevnikar Z, Obermajer N, Doljak B, et al. J Leukoc Biol. 90(1):99-109. 2011.
14. Comrie WA, Babich A, Burkhardt JK. J Cell Biol. 208(4):475-91. 2015.
15. Comrie WA, Li S, Boyle S, et al. J Cell Biol. 208(4):457-73. 2015.
16. Jankowska KI, Williamson EK, Roy NH, et al. Front Immunol. 9:25. 2018.
Indirect Elisa Protocol
Flow Cytometry
IF
IF Microscopy
Immunoprecipitation Protocol
General Western Blot Protocol

Certificate of Analysis

Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.