Table 4. Primary and secondary antibodies for detection of tubulin PTMs and modifying enzymes.
| Tubulin PTMs | |||||
|---|---|---|---|---|---|
| Modification | Name | Type | Production | References | Commercial availability |
| All α-tubulin isoforms | B-5-1-2 | Mouse Monoclonal |
Raised against Sarkosyl-resistant filaments from sea urchin sperm axonemes | (Piperno, LeDizet, & Chang, 1987) | T5168 Sigma Aldrich® |
| All β-tubulin isoforms | TUB 2.1 | Mouse Monoclonal |
Raised against rat brain tubulin; Recognizes an epitope in the C-terminal part of all five isoforms of β-tubulin (between amino acids 281-446) | (Gozes & Barnstable, 1982) | T4026 Sigma Aldrich® |
| C-terminal -GEEEGEEF and -GEEEGEEY on α-tubulin | Y1/2 | Rat Monoclonal |
Initially raised against purified yeast tubulin (-EEF) | (Kilmartin, Wright, & Milstein, 1982) | MAB1864 Millipore |
| C-terminal –GEEEGEE on α-tubulin | Anti-detyr-tubulin | Rabbit Polyclonal |
Raised against -GEEEGEE | (Gundersen, Kalnoski, & Bulinski, 1984; Paturle-Lafanechere et al., 1991) | AB320 Milipore |
| C-terminal –GEEEGEE on α-tubulin | Anti-detyr-tubulin | Rabbit Polyclonal |
Raised against 10 residue synthetic peptide of the C-terminal domain of human α-tubulin | (Berezniuk et al., 2012) | ab48389 abcam |
| C-terminal -GEEEGE on α-tubulin | Δ2-α-tubulin detyrosination | Rabbit Polyclonal |
Raised against -EGEEEGE | (Gundersen et al., 1984; Paturle-Lafanechere et al., 1994) | AB3203 Milipore |
| C-terminal – GEEEG on α-tubulin | 3EG | Rabbit Polyclonal |
Raised against -GEGEEEG | (Aillaud et al., 2016) | Not available |
| γ-Linked En side chain (n=1,2,3…) on modified E | GT335 | Mouse Monoclonal |
Raised against octapeptide EGEGE*EEG, modified by the addition of two glutamyl units onto the fifth E | (Wolff et al., 1992) | AG-20B-002 AdipoGen |
| C-terminal - En (n≥3) | 1D5 | Mouse Monoclonal |
Raised against the peptide –VDSVEGEGEEEGEE; Recognizes both detyrosinated and polyglutamylated α and β-tubulin with a minimum side chain length of 3 glutamyl residues | (Wehland & Weber, 1987) (Rudiger, Rudiger, Wehland, & Weber, 1999) |
302011 Synaptic Systems |
| C-terminal - En (n≥3) | PolyE | Rabbit Polyclonal |
Recognizes elongated side chains | (Rogowski et al., 2010; Shang, Li, & Gorovsky, 2002) | AG-25B-0030-C05 AdipoGen |
| γ-Linked G side chain (n = 1) on modified E | TAP952 | Mouse Monoclonal |
Raised against Paramecium axonemal tubulin | (Bre et al., 1996; Bre, Redeker, Vinh, Rossier, & Levilliers, 1998; Callen et al., 1994) | MABS277 Millipore |
| γ-Linked Gn side chain (n≥3) on modified E | AXO49 | Mouse Monoclonal |
Raised against Paramecium axonemal tubulin | (Bre et al., 1996; Bre et al., 1998; Callen et al., 1994) | MABS276 Millipore |
| C-terminal polyG chains | polyG | Rabbit Polyclonal |
Recognizes long polyglycylation side chains | (Rogowski et al., 2009; Shang et al., 2002) | Not available |
| Acetylation K40 on α-tubulin | 6-11B-1 | Mouse monoclonal |
Raised against flagellar tubulin of sea urchin | (LeDizet & Piperno, 1991); (Piperno & Fuller, 1985) | MABT868 Millipore |
| Phosphorylation S172 on β-tubulin | Anti-phospho-peptide P172 | Rabbit Polyclonal |
Raised against the peptide Ac-VVPpSPKVSDTVVEC-CONH2 | (Fourest-Lieuvin et al., 2006) | Not available |
| Methylation K40 on α-tubulin | α-TubK40me3 | Rabbit Polyclonal |
Raised against trimethylated K40 peptide (Ac-GQMPSD-Kme3- TIGGGDC-amide) | (Park, Chowdhury, et al., 2016; Park, Powell, et al., 2016) | Not available |
| Modifying enzymes | |||||
| Modification | Name | Type | Production | References | Commercial availability |
| Tubulin tyrosination | 2E5F8 | Mouse monoclonal |
Raised against TTL fusion protein Ag4708 | Unpublished data | 66076-1-Ig Proteintech |
| Tubulin tyrosination | Anti-TTL | Rabbit Polyclonal |
Raised against TTL fusion protein Ag4526 | (Barisic et al., 2015) | 13618-1-AP Proteintech |
| Polyglutamylation | Anti-TTLL1 | Ginea pig Polyclonal |
Raised against purified recombinant TTLL1 | (Ikegami, Sato, Nakamura, Ostrowski, & Setou, 2010) | Several commercial vendors |
| Polyglutamylation | Anti-TTLL4 | Rabbit Polyclonal |
Raised against TTLL4 peptide (aa 516-653) | (Xia et al., 2016) |
PAB22002 Abnova |
| Polyglutamylation | Anti-TTLL5 | Rabbit Polyclonal |
Raised against TTLL5 peptide (aa 1000-1088) | (Sergouniotis et al., 2014) |
PAB22614 Abnova |
| Polyglutamylation | Anti-TTLL6 | Rabbit Monoclonal |
Raised against a human TTLL6 peptide using ARM Technology | (Xia et al., 2016) | H00284076-K Abnova |
| Polyglutamylation | Anti-TTLL7 | Rabbit Polyclonal |
Raised against maltose-binding-fused TTLL7 △370; aa 371-609) | (Ikegami et al., 2006) | Several commercial vendors |
| Deglutamylation | CCP1 (LM-1A7) | Mouse Monoclonal |
Raised against recombinant AGTPBP1 protein of human origin | (Xia et al., 2016) | sc-134251 Santa Cruz Biotechnology |
| Deglutamylation | CCP2 (S-13) | Rabbit Polyclonal |
Raised against a peptide mapping near the C-terminus of CCP2 of human origin. | (Xia et al., 2016) | sc-138193 Santa Cruz Biotechnology |
| Deglutamylation | CCP5 (N-18) | Rabbit Polyclonal |
Raised against human CCP5 | (Wu, Wei, & Morgan, 2017) |
Ab118621 abcam |
| Deglutamylation | CCP6 (N-14) | Rabbit Polyclonal |
Raised against a peptide mapping near the N-terminus of CCP6 of human origin | (Li et al., 2016; Ye et al., 2014) | Discontinued, Santa Cruz Biotechnology |
| Acetylation | Anti-αTAT1 | Rabbit Polyclonal |
Recognizes residues 1-236 of αTAT1 | (Shida, Cueva, Xu, Goodman, & Nachury, 2010) | Other peptides-Several commercial vendors |
| Deacetylation | Anti-HDAC6 | Rabbit Polyclonal |
Recognizes residues 1031-1215 of HDAC6 | (Hubbert et al., 2002) | 07-732 Millipore |
| Deacetylation | Anti-SIRT2 | Rabbit Polyclonal |
Raised against synthetic peptide corresponding to Sirt2 amino acids 341-352 | (Chopra et al., 2012) | S8447 Sigma Aldrich® |
| Phosphorylation | A17 | Mouse monoclonal |
Raised against Xenopus p34cdc2 protein (C-terminal two-thirds) | (Goodger, Gannon, Hunt, & Morgan, 1996) | ab18 abcam |
| Methylation | Anti-SETD2 | Rabbit Polyclonal |
Raised against synthetic peptide from within residues 500 - 600 of human SETD2 | (Park, Powell, et al., 2016) | ab31358 abcam |