Specificity Controls for Immunocytochemistry: The Antigen Preadsorption Test Can Lead to Inaccurate Assessment of Antibody Specificity

Journal of Histochemistry and Cytochemistry

Volumn 60, Issue 3, 2012, Pages 174-187

  Holmseth, Silvia ^a   Zhou, Yun ^a   Follin Arbelet, Virginie V ^a   Lehre, Knut Petter ^a   Bergles, Dwight E ^b   Danbolt, Niels Christian ^a  

^a UNIVERSITY OF OSLO   (Norway)

^b JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

antibody dilution; blocking peptide; blot fixation test; GABA transporter; glutamate transporter; knockout mice; neoantigens

Indexed keywords

ANTIGEN; EPITOPE; 4 AMINOBUTYRIC ACID CARRIER; ANTIBODY; ANTISERUM; EXCITATORY AMINO ACID TRANSPORTER 2; EXCITATORY AMINO ACID TRANSPORTER 3; SLC1A1 PROTEIN, MOUSE; SLC6A12 PROTEIN, MOUSE;

ACCURACY; ANIMAL EXPERIMENT; ANIMAL TISSUE; ANTIBODY AFFINITY; ANTIBODY SPECIFICITY; ARTICLE; CONTROLLED STUDY; CROSS REACTION; IMMUNOADSORPTION; IMMUNOCYTOCHEMISTRY; KNOCKOUT MOUSE; MALE; MOUSE; NONHUMAN; NUCLEOTIDE SEQUENCE; PRIORITY JOURNAL; PURIFICATION; RAT; SENSITIVITY AND SPECIFICITY; TISSUE SECTION; WESTERN BLOTTING; WILD TYPE; ADSORPTION; ANIMAL; ARTIFACT; IMMUNOHISTOCHEMISTRY; IMMUNOLOGY; METABOLISM; METHODOLOGY; MOUSE MUTANT; SOLUTION AND SOLUBILITY; WISTAR RAT;

MUS;

ADSORPTION; ANIMALS; ANTIBODIES; ANTIBODY AFFINITY; ANTIBODY SPECIFICITY; ANTIGENS; ARTIFACTS; BLOTTING, WESTERN; CROSS REACTIONS; EPITOPES; EXCITATORY AMINO ACID TRANSPORTER 2; EXCITATORY AMINO ACID TRANSPORTER 3; GABA PLASMA MEMBRANE TRANSPORT PROTEINS; IMMUNE SERA; IMMUNOHISTOCHEMISTRY; MICE; MICE, KNOCKOUT; RATS; RATS, WISTAR; SENSITIVITY AND SPECIFICITY; SOLUTIONS;

EID: 84859631248     PISSN: 00221554     EISSN: None     Source Type: Journal    
DOI: 10.1369/0022155411434828     Document Type: Article

References (42)

1. Boenisch T.Heat-induced antigen retrieval: what are we retrieving?.J Histochem Cytochem. 2006;54:961-964.
2. Burry RW.Specificity controls for immunocytochemical methods.J Histochem Cytochem. 2000;48:163-166.
3. Burry RW.Controls for immunocytochemistry: an update.J Histochem Cytochem. 2011;59:6-12.
4. Couchman JR.Commercial antibodies: the good, bad, and really ugly.J Histochem Cytochem. 2009;57:7-8.
5. Danbolt NC.Glutamate uptake.Prog Neurobiol. 2001;65:1-105.
6. Danbolt NC,Lehre KP,Dehnes Y,Chaudhry FA,Levy LM.Localization of transporters using transporter-specific antibodies.Methods Enzymol. 1998;296:388-407.
7. Danbolt NC,Storm-Mathisen J,Kanner BI.An [Na++ K+]coupled L-glutamate transporter purified from rat brain is located in glial cell processes.Neuroscience. 1992;51:295-310.
8. DeGiorgio LA,Konstantinov KN,Lee SC,Hardin JA,Volpe BT,Diamond B.A subset of lupus anti-DNA antibodies cross-reacts with the NR2 glutamate receptor in systemic lupus erythematosus.Nat Med. 2001;7:1189-1193.
9. Dolman DE,Lawrenson JG,Abbott NJ.Cross reactivity of polyclonal GFAP antiserum: implications for the in-vitro characterisation of brain endothelium.Brain Res. 2004;1012:185-186.
10. Fritschy J-M.Is my antibody-staining specific? How to deal with pitfalls of immunohistochemistry.Eur J Neurosci. 2008;28:2365-2370.
11. Furness DN,Dehnes Y,Akhtar AQ,Rossi DJ,Hamann M,Grutle NJ,Gundersen V,Holmseth S,Lehre KP,Ullensvang K,Wojewodzic M,Zhou Y,Attwell D,Danbolt NC.A quantitative assessment of glutamate uptake into hippocampal synaptic terminals and astrocytes: new insights into a neuronal role for excitatory amino acid transporter 2 (EAAT2).Neuroscience. 2008;157:80-94.
12. Hoffman EM,Schechter R,Miller KE.Fixative composition alters distributions of immunoreactivity for glutaminase and two markers of nociceptive neurons, Nav1.8 and TRPV1, in the rat dorsal root ganglion.J Histochem Cytochem. 2010;58:329-344.
13. Holmseth S,Dehnes Y,Bjørnsen LP,Boulland JL,Furness DN,Bergles D,Danbolt NC.Specificity of antibodies: unexpected cross reactivity of antibodies directed against the EAAT3 (EAAC) glutamate transporter.Neuroscience. 2005;136:649-660.
14. Holmseth S,Lehre KP,Danbolt NC.Specificity controls for immunocytochemistry.Anat Embryol (Berl). 2006;211:257-266.
15. Holmseth S,Scott HA,Real K,Lehre KP,Leergaard TB,Bjaalie JG,Danbolt NC.The concentrations and distributions of three C-terminal variants of the GLT1 (EAAT2; slc1a2) glutamate transporter protein in rat brain tissue suggest differential regulation.Neuroscience. 2009;162:1055-1071.
16. James JA,McClain MT,Koelsch G,Williams DG,Harley JB.Side-chain specificities and molecular modelling of peptide determinants for two anti-Sm B/B' autoantibodies.J Autoimmun. 1999;12:43-49.
17. Josephsen K,Smith CE,Nanci A.Selective but nonspecific immunolabeling of enamel protein-associated compartments by a monoclonal antibody against vimentin.J Histochem Cytochem. 1999;47:1237-1245.
18. Kalyuzhny AE.The dark side of the immunohistochemical moon: industry.J Histochem Cytochem. 2009;57:1099-1101.
19. Lehre AC,Rowley NM,Zhou Y,Holmseth S,Guo C,Holen T,Hua R,Laake P,Olofsson AM,Poblete-Naredo I,Rusakov DA,Madsen KK,Clausen RP,Schousboe A,White HS,Danbolt NC.Deletion of the betaine-GABA transporter (BGT1; slc6a12) gene does not affect seizure thresholds of adult mice.Epilepsy Res. 2011;95:70-81.
20. Lehre KP,Danbolt NC.The number of glutamate transporter subtype molecules at glutamatergic synapses: chemical and stereological quantification in young adult rat brain.J Neurosci. 1998;18:8751-8757.
21. Lehre KP,Levy LM,Ottersen OP,Storm-Mathisen J,Danbolt NC.Differential expression of two glial glutamate transporters in the rat brain: quantitative and immunocytochemical observations.J Neurosci. 1995;15:1835-1853.
22. Liu QR,López-Corcuera B,Mandiyan S,Nelson H,Nelson N.Molecular characterization of 4 pharmacologically distinct alpha-aminobutyric acid transporters in mouse brain.J Biol Chem. 1993;268:2106-2112.
23. Lorincz A,Nusser Z.Specificity of immunoreactions: the importance of testing specificity in each method.J Neurosci. 2008;28:9083-9086.
24. Lowry OH,Roberts NR,Leiner KY,Wu ML,Farr AL,Albers RW.The quantitative histochemistry of brain, III: Ammon's horn.J Biol Chem. 1954;207:39-49.
25. Ottersen OP.Postembedding light- and electron microscopic immunocytochemistry of amino acids: description of a new model system allowing identical conditions for specificity testing and tissue processing.Exp Brain Res. 1987;69:167-174.
26. Paavilainen L,Edvinsson A,Asplund A,Hober S,Kampf C,Ponten F,Wester K.The impact of tissue fixatives on morphology and antibody-based protein profiling in tissues and cells.J Histochem Cytochem. 2010;58:237-246.
27. Peghini P,Janzen J,Stoffel W.Glutamate transporter EAAC-1-deficient mice develop dicarboxylic aminoaciduria and behavioral abnormalities but no neurodegeneration.EMBO J. 1997;16:3822-3832.
28. Pines G,Danbolt NC,Bjørås M,Zhang Y,Bendahan A,Eide L,Koepsell H,Storm-Mathisen J,Seeberg E,Kanner BI.Cloning and expression of a rat brain L-glutamate transporter.Nature. 1992;360:464-467.
29. Pool CW,Buijs RMMolecular neuroanatomy. Van LB, ed. Amsterdam: Elsevier Science Publishers B. V. Biomedical Division; 1988:.
30. Pradidarcheep W,Labruyere WT,Dabhoiwala NF,Lamers WH.Lack of specificity of commercially available antisera: better specifications needed.J Histochem Cytochem. 2008;56:1099-1111.
31. Rhodes KJ,Trimmer JS.Antibodies as valuable neuroscience research tools versus reagents of mass distraction.J Neurosci. 2006;26:8017-8020.
32. Saper CB.An open letter to our readers on the use of antibodies.J Comp Neurol. 2005;493:477-478.
33. Saper CB.A guide to the perplexed on the specificity of antibodies.J Histochem Cytochem. 2009;57:1-5.
34. Saper CB,Sawchenko PE.Magic peptides, magic antibodies: guidelines for appropriate controls for immunohistochemistry.J Comp Neurol. 2003;465:161-163.
35. Shelanski ML,Gaskin F,Cantor CR.Microtubule assembly in the absence of added nucleotides.Proc Natl Acad Sci U S A. 1973;70:765-768.
36. Sibille P,Ternynck T,Nato F,Buttin G,Strosberg D,Avrameas A.Mimotopes of polyreactive anti-DNA antibodies identified using phage-display peptide libraries.Eur J Immunol. 1997;27:1221-1228.
37. Swaab DF,Pool CW,Van Leeuwen FW.Can specificity ever be proved in immunocytochemical staining.J Histochem Cytochem. 1977;25:388-391.
38. Tanaka K,Watase K,Manabe T,Yamada K,Watanabe M,Takahashi K,Iwama H,Nishikawa T,Ichihara N,Hori S,Takimoto M,Wada K.Epilepsy and exacerbation of brain injury in mice lacking the glutamate transporter GLT-1.Science. 1997;276:1699-1702.
39. Webster JD,Miller MA,Dusold D,Ramos-Vara J.Effects of prolonged formalin fixation on diagnostic immunohistochemistry in domestic animals.J Histochem Cytochem. 2009;57:753-761.
40. Willingham MC.Conditional epitopes: is your antibody always specific?.J Histochem Cytochem. 1999;47:1233-1236.
41. Xie R,Chung J-Y,Ylaya K,Williams RL,Guerrero N,Nakatsuka N,Badie C,Hewitt SM.Factors influencing the degradation of archival formalin-fixed paraffin-embedded tissue sections.J Histochem Cytochem. 2011;59:356-365.
42. Zhou Y,Holmseth S,Hua R,Lehre AC,Olofsson AM,Poblete-Naredo I,Kempson SA,Danbolt NC.The betaine-GABA transporter (BGT1, slc6a12) is predominantly expressed in the liver and at lower levels in the kidneys and at the brain surface.Am J Physiol Renal Physiol. 2012;302:F316-328.