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What proteins stick to ProteinG on preclearing?


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#1 BIOKMST

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Posted 17 September 2009 - 11:47 AM

Does anybody know what proteins have a tendency to bind to Protein G beads when preclearing lysate for an IP?

Thanks!

Cindy

#2 dpo

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Posted 24 September 2009 - 03:16 AM

From Miles et al., Mol Cell Prot 2005


Supplemental Table 1. Analysis of IgG control lane by tandem mass spectrometry.

# Uniprot Protein MW Predicted % error
1 O14802 DNA-directed RNA polymerase III largest subunit 155749 0.0
2 Q03468 DNA excision repair protein ERCC-6 168416 0.0
3 P54252 Machado-Joseph disease protein 1 43450 0.0
4 O60229 Huntingtin-associated protein-interacting protein 192229 0.4
5 Q8NA23 WD-repeat protein 31 40840 0.8
6 O00469 Procollagen-lysine,2-oxoglutarate 5-dioxygenase 2 precursor 84664 0.9
7 * P51805 Plexin A3 precursor 207663 1.1
8 P14410 Sucrase-isomaltase, intestinal 209273 1.4
9 Q92879 CUG triplet repeat RNA-binding protein 1 52063 1.4
10 Q9UBI9 Headcase protein homolog 58837 1.5
11 Q9Y490 Talin 1 269718 1.5
12 Q96KP1 Exocyst complex component Sec5 104066 1.7
13 * Q13936 Voltage-dependent L-type calcium channel alpha-1C subunit 248887 1.9
14 Q96KX2 F-actin capping protein alpha-3 subunit 35025 1.9
15 Q13685 Angio-associated migratory cell protein 49016 2.3
16 Q9H777 Zinc phosphodiesterase ELAC protein 1 40019 2.5
17 P49321 Nuclear autoantigenic sperm protein 85238 2.5
18 Q9Y2T1 Axin 2 93558 3.1
19 Q96QZ0 Pannexin 3 44683 3.1
20 Q96PX8 SLIT and NTRK-like protein 1 precursor 77735 3.5
21 P49792 Ran-binding protein 2 358218 3.5
22 Q92628 Hypothetical protein KIAA0232 155620 3.7
23 O75891 10-formyltetrahydrofolate dehydrogenase 98829 3.8
24 * P04264 Keratin, type II cytoskeletal 1 65886 3.8
25 Q9H2X0 Chordin precursor 102014 4.4
26 * Q9H7Z3 Protein C14orf102 132417 4.4
27 Q01831 DNA-repair protein complementing XP-C cells 105850 4.4
28 P01771 Ig heavy chain V-III region HIL 13566 4.9
29 * P12035 Keratin, type II cytoskeletal 3 64511 4.9
30 P52849 Heparin sulfate N-deacetylase/N-sulfotransferase 100874 5.0
31 Q04695 Keratin, type I cytoskeletal 17 (Cytokeratin 17 47974 5.3
32 P28358 Homeobox protein Hox-D10 (Hox-4D 38509 5.3
33 P19367 Hexokinase, type I 102502 6.1
34 Q01101 Zinc finger protein IA-1 (Insulinoma-associated protein 1) 52923 6.1
35 Q8WUF5 RelA-associated inhibitor (Inhibitor of ASPP protein 44106 6.2
36 * Q99456 Keratin, type I cytoskeletal 12 (Cytokeratin 12 53511 6.5
37 Q9UK39 Nocturnin (CCR4 protein homolog) 48151 7.0
38 P54819 Adenylate kinase isoenzyme 2, mitochondrial 26346 7.1
39 P98164 Low-density lipoprotein receptor-related protein 2 precursor (Megalin 521928 7.1
40 Q9NRM7 Serine/threonine protein kinase LATS2 120193 7.4
41 Q8N653 Leucine-zipper-like transcriptional regulator 1 (LZTR-1) 94718 7.4
42 * Q8IUG5 Myosin XVIIIB (Myosin 18B) 285183 8.3
43 O00754 Lysosomal alpha-mannosidase precursor 113672 9.3
44 P43361 Melanoma-associated antigen 8 (MAGE-8 antigen) 25197 9.4

Edited by dpo, 24 September 2009 - 03:16 AM.


#3 BIOKMST

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Posted 29 September 2009 - 09:17 AM

From Miles et al., Mol Cell Prot 2005


Supplemental Table 1. Analysis of IgG control lane by tandem mass spectrometry.

# Uniprot Protein MW Predicted % error
1 O14802 DNA-directed RNA polymerase III largest subunit 155749 0.0
2 Q03468 DNA excision repair protein ERCC-6 168416 0.0
3 P54252 Machado-Joseph disease protein 1 43450 0.0
4 O60229 Huntingtin-associated protein-interacting protein 192229 0.4
5 Q8NA23 WD-repeat protein 31 40840 0.8
6 O00469 Procollagen-lysine,2-oxoglutarate 5-dioxygenase 2 precursor 84664 0.9
7 * P51805 Plexin A3 precursor 207663 1.1
8 P14410 Sucrase-isomaltase, intestinal 209273 1.4
9 Q92879 CUG triplet repeat RNA-binding protein 1 52063 1.4
10 Q9UBI9 Headcase protein homolog 58837 1.5
11 Q9Y490 Talin 1 269718 1.5
12 Q96KP1 Exocyst complex component Sec5 104066 1.7
13 * Q13936 Voltage-dependent L-type calcium channel alpha-1C subunit 248887 1.9
14 Q96KX2 F-actin capping protein alpha-3 subunit 35025 1.9
15 Q13685 Angio-associated migratory cell protein 49016 2.3
16 Q9H777 Zinc phosphodiesterase ELAC protein 1 40019 2.5
17 P49321 Nuclear autoantigenic sperm protein 85238 2.5
18 Q9Y2T1 Axin 2 93558 3.1
19 Q96QZ0 Pannexin 3 44683 3.1
20 Q96PX8 SLIT and NTRK-like protein 1 precursor 77735 3.5
21 P49792 Ran-binding protein 2 358218 3.5
22 Q92628 Hypothetical protein KIAA0232 155620 3.7
23 O75891 10-formyltetrahydrofolate dehydrogenase 98829 3.8
24 * P04264 Keratin, type II cytoskeletal 1 65886 3.8
25 Q9H2X0 Chordin precursor 102014 4.4
26 * Q9H7Z3 Protein C14orf102 132417 4.4
27 Q01831 DNA-repair protein complementing XP-C cells 105850 4.4
28 P01771 Ig heavy chain V-III region HIL 13566 4.9
29 * P12035 Keratin, type II cytoskeletal 3 64511 4.9
30 P52849 Heparin sulfate N-deacetylase/N-sulfotransferase 100874 5.0
31 Q04695 Keratin, type I cytoskeletal 17 (Cytokeratin 17 47974 5.3
32 P28358 Homeobox protein Hox-D10 (Hox-4D 38509 5.3
33 P19367 Hexokinase, type I 102502 6.1
34 Q01101 Zinc finger protein IA-1 (Insulinoma-associated protein 1) 52923 6.1
35 Q8WUF5 RelA-associated inhibitor (Inhibitor of ASPP protein 44106 6.2
36 * Q99456 Keratin, type I cytoskeletal 12 (Cytokeratin 12 53511 6.5
37 Q9UK39 Nocturnin (CCR4 protein homolog) 48151 7.0
38 P54819 Adenylate kinase isoenzyme 2, mitochondrial 26346 7.1
39 P98164 Low-density lipoprotein receptor-related protein 2 precursor (Megalin 521928 7.1
40 Q9NRM7 Serine/threonine protein kinase LATS2 120193 7.4
41 Q8N653 Leucine-zipper-like transcriptional regulator 1 (LZTR-1) 94718 7.4
42 * Q8IUG5 Myosin XVIIIB (Myosin 18B) 285183 8.3
43 O00754 Lysosomal alpha-mannosidase precursor 113672 9.3
44 P43361 Melanoma-associated antigen 8 (MAGE-8 antigen) 25197 9.4


Thanks for the reply, but these are not correct. These proteins are IgG binding proteins. I would like to know all proteins binding to Protein G, without antibody binding.

Best,
Cindy

#4 Dr Teeth

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Posted 29 September 2009 - 09:50 AM

From Miles et al., Mol Cell Prot 2005


Supplemental Table 1. Analysis of IgG control lane by tandem mass spectrometry.

# Uniprot Protein MW Predicted % error
1 O14802 DNA-directed RNA polymerase III largest subunit 155749 0.0
2 Q03468 DNA excision repair protein ERCC-6 168416 0.0
3 P54252 Machado-Joseph disease protein 1 43450 0.0
4 O60229 Huntingtin-associated protein-interacting protein 192229 0.4
5 Q8NA23 WD-repeat protein 31 40840 0.8
6 O00469 Procollagen-lysine,2-oxoglutarate 5-dioxygenase 2 precursor 84664 0.9
7 * P51805 Plexin A3 precursor 207663 1.1
8 P14410 Sucrase-isomaltase, intestinal 209273 1.4
9 Q92879 CUG triplet repeat RNA-binding protein 1 52063 1.4
10 Q9UBI9 Headcase protein homolog 58837 1.5
11 Q9Y490 Talin 1 269718 1.5
12 Q96KP1 Exocyst complex component Sec5 104066 1.7
13 * Q13936 Voltage-dependent L-type calcium channel alpha-1C subunit 248887 1.9
14 Q96KX2 F-actin capping protein alpha-3 subunit 35025 1.9
15 Q13685 Angio-associated migratory cell protein 49016 2.3
16 Q9H777 Zinc phosphodiesterase ELAC protein 1 40019 2.5
17 P49321 Nuclear autoantigenic sperm protein 85238 2.5
18 Q9Y2T1 Axin 2 93558 3.1
19 Q96QZ0 Pannexin 3 44683 3.1
20 Q96PX8 SLIT and NTRK-like protein 1 precursor 77735 3.5
21 P49792 Ran-binding protein 2 358218 3.5
22 Q92628 Hypothetical protein KIAA0232 155620 3.7
23 O75891 10-formyltetrahydrofolate dehydrogenase 98829 3.8
24 * P04264 Keratin, type II cytoskeletal 1 65886 3.8
25 Q9H2X0 Chordin precursor 102014 4.4
26 * Q9H7Z3 Protein C14orf102 132417 4.4
27 Q01831 DNA-repair protein complementing XP-C cells 105850 4.4
28 P01771 Ig heavy chain V-III region HIL 13566 4.9
29 * P12035 Keratin, type II cytoskeletal 3 64511 4.9
30 P52849 Heparin sulfate N-deacetylase/N-sulfotransferase 100874 5.0
31 Q04695 Keratin, type I cytoskeletal 17 (Cytokeratin 17 47974 5.3
32 P28358 Homeobox protein Hox-D10 (Hox-4D 38509 5.3
33 P19367 Hexokinase, type I 102502 6.1
34 Q01101 Zinc finger protein IA-1 (Insulinoma-associated protein 1) 52923 6.1
35 Q8WUF5 RelA-associated inhibitor (Inhibitor of ASPP protein 44106 6.2
36 * Q99456 Keratin, type I cytoskeletal 12 (Cytokeratin 12 53511 6.5
37 Q9UK39 Nocturnin (CCR4 protein homolog) 48151 7.0
38 P54819 Adenylate kinase isoenzyme 2, mitochondrial 26346 7.1
39 P98164 Low-density lipoprotein receptor-related protein 2 precursor (Megalin 521928 7.1
40 Q9NRM7 Serine/threonine protein kinase LATS2 120193 7.4
41 Q8N653 Leucine-zipper-like transcriptional regulator 1 (LZTR-1) 94718 7.4
42 * Q8IUG5 Myosin XVIIIB (Myosin 18B) 285183 8.3
43 O00754 Lysosomal alpha-mannosidase precursor 113672 9.3
44 P43361 Melanoma-associated antigen 8 (MAGE-8 antigen) 25197 9.4


Thanks for the reply, but these are not correct. These proteins are IgG binding proteins. I would like to know all proteins binding to Protein G, without antibody binding.

Best,
Cindy


If the above proteins bind to IgG precipitated with protein G beads, then the proteins that bind to protein G in the absence of IgG should be a subset of the above.

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley




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