Reference sera for antinuclear antibodies: II. Further definition of antibody specificities in international antinuclear antibody reference sera by immunofluorescence and western blotting

Josef S. Smolen, Brian Butcher, Marvin J. Fritzler, Thomas Gordon, John Hardin, Joachim R. Kalden, Robert Lahita, Ravinder N. Maini, Westley Reeves, Morris Reichlin, Naomi Rothfield, Yoshinari Takasaki, Walther J. Van Venrooij, Eng M. Tan

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Objective. To define the fine specificity of the 10 reference sera used for determination of antinuclear antibodies (ANA) and ANA subsets which are available from the Arthritis Foundation (AF) and from the Centers for Disease Control and Prevention (CDC). Methods. AF/CDC sera were assessed by experienced laboratory staff, using indirect immunofluorescence and Western blotting. Results. The original assignment of fluorescence patterns to 4 reference sera was confirmed, and the fluorescence intensities were determined using reference fluorescent beads. On Western blots, sera AF/CDC2 (anti-SS-B/La) and AF/CDC7 (anti-SS-A/Ro) did not detect Ro antigens, sera AF/CDC9 and AF/CDC10 appeared to be monospecific anti-Scl-70 and anti-Jo-1 sera, respectively, serum AF/CDC4 (anti-U1 small nuclear RNP) recognized the 70-kd band, and serum AF/CDC5 recognized the Sm antigen with its multiple bands. Semiquantitative analyses revealed that AF/CDC5, AF/CDC2, and AF/CDC10 were strongly reactive sera, whereas AF/CDC4 and AF/CDC9 were much weaker and should be used at lower dilutions on Western blots. Conclusion. The AF/CDC ANA reference sera, originally described as reference reagents for indirect immunofluorescence and double immunodiffusion techniques, are also useful for Western blotting. The data presented herein further support the use of these sera far reference purposes.

Original languageEnglish (US)
Pages (from-to)413-418
Number of pages6
JournalArthritis and Rheumatism
Volume40
Issue number3
DOIs
StatePublished - 1997
Externally publishedYes

Fingerprint

Antibody Specificity
Antinuclear Antibodies
Arthritis
Fluorescent Antibody Technique
Western Blotting
Serum
Centers for Disease Control and Prevention (U.S.)
Indirect Fluorescent Antibody Technique
Fluorescence
Small Nuclear Ribonucleoproteins
Immunodiffusion

ASJC Scopus subject areas

  • Immunology
  • Rheumatology

Cite this

Reference sera for antinuclear antibodies : II. Further definition of antibody specificities in international antinuclear antibody reference sera by immunofluorescence and western blotting. / Smolen, Josef S.; Butcher, Brian; Fritzler, Marvin J.; Gordon, Thomas; Hardin, John; Kalden, Joachim R.; Lahita, Robert; Maini, Ravinder N.; Reeves, Westley; Reichlin, Morris; Rothfield, Naomi; Takasaki, Yoshinari; Van Venrooij, Walther J.; Tan, Eng M.

In: Arthritis and Rheumatism, Vol. 40, No. 3, 1997, p. 413-418.

Research output: Contribution to journalArticle

Smolen, JS, Butcher, B, Fritzler, MJ, Gordon, T, Hardin, J, Kalden, JR, Lahita, R, Maini, RN, Reeves, W, Reichlin, M, Rothfield, N, Takasaki, Y, Van Venrooij, WJ & Tan, EM 1997, 'Reference sera for antinuclear antibodies: II. Further definition of antibody specificities in international antinuclear antibody reference sera by immunofluorescence and western blotting', Arthritis and Rheumatism, vol. 40, no. 3, pp. 413-418. https://doi.org/10.1002/art.1780400304
Smolen, Josef S. ; Butcher, Brian ; Fritzler, Marvin J. ; Gordon, Thomas ; Hardin, John ; Kalden, Joachim R. ; Lahita, Robert ; Maini, Ravinder N. ; Reeves, Westley ; Reichlin, Morris ; Rothfield, Naomi ; Takasaki, Yoshinari ; Van Venrooij, Walther J. ; Tan, Eng M. / Reference sera for antinuclear antibodies : II. Further definition of antibody specificities in international antinuclear antibody reference sera by immunofluorescence and western blotting. In: Arthritis and Rheumatism. 1997 ; Vol. 40, No. 3. pp. 413-418.
@article{4389958800014d47827411eac0c110a0,
title = "Reference sera for antinuclear antibodies: II. Further definition of antibody specificities in international antinuclear antibody reference sera by immunofluorescence and western blotting",
abstract = "Objective. To define the fine specificity of the 10 reference sera used for determination of antinuclear antibodies (ANA) and ANA subsets which are available from the Arthritis Foundation (AF) and from the Centers for Disease Control and Prevention (CDC). Methods. AF/CDC sera were assessed by experienced laboratory staff, using indirect immunofluorescence and Western blotting. Results. The original assignment of fluorescence patterns to 4 reference sera was confirmed, and the fluorescence intensities were determined using reference fluorescent beads. On Western blots, sera AF/CDC2 (anti-SS-B/La) and AF/CDC7 (anti-SS-A/Ro) did not detect Ro antigens, sera AF/CDC9 and AF/CDC10 appeared to be monospecific anti-Scl-70 and anti-Jo-1 sera, respectively, serum AF/CDC4 (anti-U1 small nuclear RNP) recognized the 70-kd band, and serum AF/CDC5 recognized the Sm antigen with its multiple bands. Semiquantitative analyses revealed that AF/CDC5, AF/CDC2, and AF/CDC10 were strongly reactive sera, whereas AF/CDC4 and AF/CDC9 were much weaker and should be used at lower dilutions on Western blots. Conclusion. The AF/CDC ANA reference sera, originally described as reference reagents for indirect immunofluorescence and double immunodiffusion techniques, are also useful for Western blotting. The data presented herein further support the use of these sera far reference purposes.",
author = "Smolen, {Josef S.} and Brian Butcher and Fritzler, {Marvin J.} and Thomas Gordon and John Hardin and Kalden, {Joachim R.} and Robert Lahita and Maini, {Ravinder N.} and Westley Reeves and Morris Reichlin and Naomi Rothfield and Yoshinari Takasaki and {Van Venrooij}, {Walther J.} and Tan, {Eng M.}",
year = "1997",
doi = "10.1002/art.1780400304",
language = "English (US)",
volume = "40",
pages = "413--418",
journal = "Arthritis and Rheumatology",
issn = "2326-5191",
publisher = "John Wiley and Sons Ltd",
number = "3",

}

TY - JOUR

T1 - Reference sera for antinuclear antibodies

T2 - II. Further definition of antibody specificities in international antinuclear antibody reference sera by immunofluorescence and western blotting

AU - Smolen, Josef S.

AU - Butcher, Brian

AU - Fritzler, Marvin J.

AU - Gordon, Thomas

AU - Hardin, John

AU - Kalden, Joachim R.

AU - Lahita, Robert

AU - Maini, Ravinder N.

AU - Reeves, Westley

AU - Reichlin, Morris

AU - Rothfield, Naomi

AU - Takasaki, Yoshinari

AU - Van Venrooij, Walther J.

AU - Tan, Eng M.

PY - 1997

Y1 - 1997

N2 - Objective. To define the fine specificity of the 10 reference sera used for determination of antinuclear antibodies (ANA) and ANA subsets which are available from the Arthritis Foundation (AF) and from the Centers for Disease Control and Prevention (CDC). Methods. AF/CDC sera were assessed by experienced laboratory staff, using indirect immunofluorescence and Western blotting. Results. The original assignment of fluorescence patterns to 4 reference sera was confirmed, and the fluorescence intensities were determined using reference fluorescent beads. On Western blots, sera AF/CDC2 (anti-SS-B/La) and AF/CDC7 (anti-SS-A/Ro) did not detect Ro antigens, sera AF/CDC9 and AF/CDC10 appeared to be monospecific anti-Scl-70 and anti-Jo-1 sera, respectively, serum AF/CDC4 (anti-U1 small nuclear RNP) recognized the 70-kd band, and serum AF/CDC5 recognized the Sm antigen with its multiple bands. Semiquantitative analyses revealed that AF/CDC5, AF/CDC2, and AF/CDC10 were strongly reactive sera, whereas AF/CDC4 and AF/CDC9 were much weaker and should be used at lower dilutions on Western blots. Conclusion. The AF/CDC ANA reference sera, originally described as reference reagents for indirect immunofluorescence and double immunodiffusion techniques, are also useful for Western blotting. The data presented herein further support the use of these sera far reference purposes.

AB - Objective. To define the fine specificity of the 10 reference sera used for determination of antinuclear antibodies (ANA) and ANA subsets which are available from the Arthritis Foundation (AF) and from the Centers for Disease Control and Prevention (CDC). Methods. AF/CDC sera were assessed by experienced laboratory staff, using indirect immunofluorescence and Western blotting. Results. The original assignment of fluorescence patterns to 4 reference sera was confirmed, and the fluorescence intensities were determined using reference fluorescent beads. On Western blots, sera AF/CDC2 (anti-SS-B/La) and AF/CDC7 (anti-SS-A/Ro) did not detect Ro antigens, sera AF/CDC9 and AF/CDC10 appeared to be monospecific anti-Scl-70 and anti-Jo-1 sera, respectively, serum AF/CDC4 (anti-U1 small nuclear RNP) recognized the 70-kd band, and serum AF/CDC5 recognized the Sm antigen with its multiple bands. Semiquantitative analyses revealed that AF/CDC5, AF/CDC2, and AF/CDC10 were strongly reactive sera, whereas AF/CDC4 and AF/CDC9 were much weaker and should be used at lower dilutions on Western blots. Conclusion. The AF/CDC ANA reference sera, originally described as reference reagents for indirect immunofluorescence and double immunodiffusion techniques, are also useful for Western blotting. The data presented herein further support the use of these sera far reference purposes.

UR - http://www.scopus.com/inward/record.url?scp=0030949749&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030949749&partnerID=8YFLogxK

U2 - 10.1002/art.1780400304

DO - 10.1002/art.1780400304

M3 - Article

C2 - 9082926

AN - SCOPUS:0030949749

VL - 40

SP - 413

EP - 418

JO - Arthritis and Rheumatology

JF - Arthritis and Rheumatology

SN - 2326-5191

IS - 3

ER -