S1. Supplementary tables 1: Proteomic analysis of fetal bovine serum (FBS)
Name of the protein
Molecular weight
(KDa)
25.1
Type of protein
Structural protein 1 Crystallin, beta A1
2 Crystallin, beta A1 subunit 13.1
3 Retinol binding protein, interstitial 39.6
Structural protein
Glycoprotein
4 Bradykinin 1.1
Nonapeptide/ improves the quality
of cryopreserved semen [1]
5
Beaded filament structural protein
2, phakinin (bovine)
6 Crystallin, gamma B
7
Aldehyde dehydrogenase 3 family, member A1
Tyrosine 3-
8 monooxygenase/tryptophan 5monooxygenase activation protein, beta
9
Crystallin, gamma S c-terminal domain
10
Versican V3 splice-variant precursor
45.5
10.3
26.7
27.8
10.3
74.7
Intermediate filament
Structural protein
Enzyme
Enzyme/suppresses several
apoptotic pathways [2,3]
Structural protein
Proteoglycan
35.4 Structural protein 11 Crystallin, zeta
12 AE binding protein 1
13
Chaperonin containing TCP1, subunit 8 (theta), partial
82.3
19.8
DNA binding protein
Protein chaperone
14 Heat shock 70KDa, chain A 41.2
20.4
Chaperone protein cytoprotective properties explained by its anti-
apoptotic function [4]
Glycoprotein 15 Fibronectin variable region
16
Creatine kinase, bovine retinal chain a
17
Crystallin, gamma A (c18s mutant of bovine)
42.6
20.9
Enzyme
Structural protein
18
19
Filamin A interacting protein 1, partial
Crystallin, alpha B
20 Crystallin, alpha A (Bos taurus)
53.1
20.0
19.8
Globular protein/inhibits apoptosis induced by the mechanical forces
[5]
Structural protein
Structural protein
113.3 Membrane protein 21 Contactin 1, precursor
22
Beaded filament structural protein
2, phakinin (Bos taurus)
23 Retinal dehydrogenase 1
24 Ezrin
45.9
54.8
68.7
Lens structural protein
Enzyme
Membrane protein

25
Retinol-binding protein 3
(precursor)
Serpin peptidase inhibitor, clade F
26
(alpha-2 antiplasmin, pigment epithelium derived factor), member
1 (precursor)
27
Protein disulfide isomerase family
A, member 6 (precursor)
28 Transthyretin (precursor)
139.6
46.2
57.2
15.7
29 Clusterin preproprotein
30 Crystallin, beta A2
31 Crystallin, beta B1
32
Phosphatidylinositol-specific phospholipase D (precursor)
33
Spondin 1, extracellular matrix protein (precursor)
51.1
22.2
28.1
92.5
90.9
34 Elongation factor 1 alpha 13.0
35 Tubulin, alpha 1 10.4
36 Elongation factor 1 gamma
37
Heat shock protein 90KDa beta, partial
38
Glutamic-oxaloacetic transaminase
2, cytoplasmic
39 Transketolase
40 Crystallin, beta A4
41 Decorin, 2b
42 Creatine kinase, muscle
17.2
27.8
46.4
9.6
22.5
36.3
43.0
43 Heat shock protein 90KDa alpha 84.7
44
UDP-GlcNAc:betaGal beta-1,3-Nacetylglucosaminyltransferase 6
45
Rab geranylgeranyltransferase, alpha subunit
46 Crystallin, gamma C
47 Triosephosphate isomerase
48 Complement factor B, partial
49 Albumin, partial [Bos indicus]
50 Phosphoglycerate kinase 1
51
Secreted frizzled-related protein 2 precursor
43.8
38.1
20.9
26.7
9.1
53.9
44.5
33.3
Carrier protein
Secreted protein/plays antiapoptotic role by inhibiting lysosomal degradation of Bcl-xL
[6]
Enzyme
Thyroxine binding pre-albumin
Glycoprotein/inhibits apoptosis by
interacting with activated Bax [7]
Structural protein
Structural protein
Enzyme
Secreted protein
Enzymatic delivery of aminoacyl tRNAs to the ribosome and nuclear export of proteins
Globular protein/binds to dynein in sperm and forms cold resistant
microtubules [8]
Enzymatic delivery of aminoacyl tRNAs to the ribosome
Chaperone protein
Enzyme
Enzyme
Structural protein
Proteoglycan
Enzyme
Chaperone protein protect cells
from ROS [9], involved in freezing
resistance of bull sperm [10]
Enzyme
Enzyme
Structural protein
Enzyme
Glycoprotein
Globular protein/replaces FBS for
cryopreservation of cells [11]
Enzyme
Glycoprotein

52 Carbonic anhydrase III
53 NSFL1 (p97) cofactor (p47)
54 Tubulin, alpha 4a
55 Enolase 3 (beta, muscle)
29.4
40.6
49.6
47.1
56 Phosphorylase, glycogen; brain
57 GPD1, partial
58
Glyceraldehyde-3-phosphate dehydrogenase
59 Vimentin
96.3
37.0
24.2
53.7
60 Selenium binding protein 1
61
Leucine-rich alpha-2-glycoprotein
1 precursor
62 Keratin 7, type II
52.5
38.3
51.5
63
Protein HP-25 homolog 2 precursor
22.9
64 Alpha-1-B glycoprotein precursor 53.5
65
Glyceraldehyde-3-phosphate dehydrogenase pseudogene 2
66 Phosphoglucomutase 5 partial
35.8
45.1
67 Gelsolin 80.7
68 Phosphofructokinase, liver
69 Complement component 3
70 Keratin 5, type II
85.2
187.1
62.6
71 Fibulin-1 precursor
Ribonuclease L (2',5'-
72 oligoisoadenylate synthetasedependent)
73
Inter-alpha-trypsin inhibitor heavy chain 2 precursor
74
CAP, adenylate cyclase-associated protein, 2
75 Uridine phosphorylase 1
77.5
81.7
106.1
53.7
33.9
76
Dickkopf WNT signaling pathway inhibitor 3precursor
80 Biotinidase precursor
38.4
77 Albumin
78
Fumarylacetoacetate hydrolase
(fumarylacetoacetase)
69.2
46.1
79 Quiescin Q6 sulfhydryl oxidase 1 62.9
58.3
81 FK506 binding protein 1A, 12KDa 11.8
Enzyme
ATPase
Globular protein
Enzyme/efficiently replaces DMSO for rat hepatocytes cryopreservation
without cellular toxicity [12]
Enzyme
Enzyme
Enzyme
Intermediate filament protein
Enzyme
Glycoprotein
Cytoskeletal protein
Hibernation specific plasma protein
Glycoprotein
Enzyme
Enzyme
Lysophosphatidic acid transport protein/control and execution of
apoptosis [13] with anti-oxidant properties [14]
Enzyme
Glycoprotein
Cytoskeletal protein
Calcium binding glycol protein
Enzyme
Carrier protein
Enzyme
Enzyme
Secreted protein
Globular protein
Enzyme
Enzyme
Enzyme
Enzyme

82
Leucine aminopeptidase 3
(unligated)
83 Crystallin, beta A1
52.6
20.7
84 Alpha-2-macroglobulin precursor 167.5
85 Secernin 1
86 Heat shock 70KDa protein 4
89 Keratin 14, type I
46.1
94.4
87
Tubulin-colchicine-phomopsin A, chain D
49.9
88 Phosphogluconate dehydrogenase 53.0
51.8
109.1 90 Kinesin family member 5C
Crystal structure of human STAM1
91 VHS domain in complex with ubiquitin, chain g
Ca Model Of Bovine TricCCT
92
DERIVED FROM A 4.0
ANGSTROM Cryo-Em Map, chain Q
93 Crystallin, alpha A (bovine)
94 Transcriptional regulator ICP4
8.3
55.7
11.6
141.7
95 Filamin A, alpha
96
Nucleosome assembly protein 1like 4
275.4
44.0
99.3 97 Actinin, alpha 2, partial
98
Sortilin-related VPS10 domain containing receptor 1
99 T-complex 1, zeta
100 Complement factor B precursor
129.9
53.5
85.3
101 Heat shock protein 90KDa beta 78.6
102 Endopin 1b-like 26.3
103 Tight junction protein (ZO) 1
104
EGF-like, fibronectin type III and laminin G domains
105 Keratin 10, type I
106
Oligodendrocyte myelin glycoprotein
144.0
99.6
54.5
49.4
Enzyme
Structural protein
Globular protein/proteinase inhibitor and anti-apoptotic protein
[15]
Exocytosis regulating protein
Chaperone protein
Globular protein
Enzyme
Cytoskeletal protein
Motor protein
Trafficking protein
Chaperone protein
Structural protein
Nuclear protein
Globular protein/protect the cells
from force induced apoptosis [16]
Interact with both core and linker histones
Actin binding protein regulate cell survival by TNF-alpha-induced
survival signalling [17]
Associates with serum lipoproteins
Molecular chaperone
Glycoprotein
Chaperone protein/protect cells
from ROS [9], involved in freezing
resistance of bull sperm [10]
Serpin protease inhibitors/inhibits cysteine proteases and elastase-like serine proteases therefore,
eventually inhibiting apoptosis [18]
Transmembrane protein
Glycoprotein
Cytoskeletal protein
Glycoprotein

107
Protein phosphatase 2, regulatory subunit A, alpha
65.2
108 Hemicentin 1 612.2
109 Tripeptidyl peptidase I precursor 61.2
110 Apolipoprotein A-I like 23.6
111
Fatty acid binding protein 5, epidermal
112 N-myc downstream regulated 1
113 RAN binding protein 6
114
CAP, adenylate cyclase-associated protein 1 like
115 BRCA1 cofactor
116
Actin, gamma 2, smooth muscle, enteric
117
Malic enzyme 1, NADP(+)dependent, cytosolic
118 ATPase, class V (ER)
119 Talin 1
120 Ankyrin 2, neuronal like
121
ABI family, member 3 (NESH) binding protein
122 Keratin 1, type II
123
Eukaryotic translation initiation factor 3, subunit F pseudogene 2
124 Unnamed protein product
11.9
41.6
47.3
51.3
87.6
39.7
58.9
89.2
269.7
433.4
39.3
63.1
35.6
47.2
125 Karyopherin (importin) beta 1 97.2
126 Clathrin, heavy chain
127
Dynein, cytoplasmic 1, heavy chain 1
128
NAD(P)H dehydrogenase, quinone
2
129 Keratin 1b, type II
130
Protein tyrosine phosphatase, receptor-type, Z polypeptide 1
131
RNA polymerase, sigma-24 subunit
132
Ceruloplasmin (ferroxidase) precursor
Model Refined Against Symmetry-
133 free Cryo-em Map Of Tric-adp,
Chain P
186.8
531.6
55.5
50.0
168.4
22.8
120.7
55.0
Enzyme
Immunoglobulin superfamily
Enzyme
High density lipoprotein/promote proliferation and inhibits apoptosis
of cells [19]
Intracellular binding protein
Helix-loop-helix protein
Nuclear transport receptor
Enzyme
Induce large-scale chromatin reorganization
Globular protein
Enzyme
Enzyme
Actin binding protein
Integral membrane protein
Enzyme
Cytoskeletal protein
Involved in initiation of translation
Protein involved in transport of molecules between cytoplasm and nucleus
Involved in membrane traffic pathways
Motor protein
Enzyme
Cytoskeletal protein
Enzyme
Enzyme
Enzyme
Chaperone protein

134 Glucose-6-phosphate isomerase
135 Profilin, chain A
136
Group-specific component
(vitamin D binding protein)
137 Thioredoxin
62.8
41.6
53.3
11.8
138 Alpha-2-macroglobulin variant 22 77.3
139 Heat shock 70KDa protein, partial 69.4
140 Hypothetical protein A7H1H_1263 38.4
141 tRNA methyltransferase 5 57.2
142 Contactin 4 113.2
124.0 143 Fibulin 2
144 DCC netrin 1 receptor
145 Complement component 4A
146
Amyloid beta (A4) precursor protein-binding
147 Crystallin, beta S
148
Eukaryotic translation initiation factor 4A1
149 Transferrin
141.1
187.7
76.6
20.9
45.0
77.6
150 Fibronectin 239.3
151 Glutathione S-transferase mu 1 21.1
152 Neuronal cell adhesion molecule 130.8
153 Filamin C 277.2
154 Lumican 38.7
155 Keratin 2, type II
156 Tubulin, alpha 1c
157
Cullin-associated and neddylationdissociated 1
158
Eukaryotic translation initiation factor 3, subunit L
159
Hsc70-interacting protein isoform
X1
160
Protein kinase C and casein kinase substrate in neurons 2
161 Plexin B2
162 Ankyrin 2, neuronal
163
Superoxide dismutase 3, extracellular
64.1
46.0
123.8
61.0
40.2
51.4
204.8
363.9
26.2
Enzyme
Actin binding protein
Globular protein
Enzyme
Globular protein/proteinase inhibitor and anti- apoptotic protein
[15]
Chaperone protein
Globular protein
Enzyme
Membrane protein
Calcium binding glycol protein
Transmembrane protein
Glycoprotein
Adaptor protein localized in the nucleus
Structural protein
Enzyme
Glycoprotein
Glycoprotein/induce cell
proliferation, inhibit apoptosis [20],
improves fertility rate of
cryopreserved sperm [21]
Enzyme
Glycoprotein
Globular protein
Proteoglycan/promote cell proliferation and regulates
apoptosis [22]
Cytoskeletal protein
Globular protein
Enzyme
Initiation of protein synthesis
Chaperone protein
Enzyme
Semaphorin membrane receptor protein
Integral membrane protein
Enzyme/antioxidant and helps in reducing ROS and delaying

164 Follistatin-like 4
165 Dihydropyrimidinase-like 2
166 Pyruvate kinase, muscle
167 Phosphorylase, glycogen, liver
168 Actinin, alpha 1
78.8
58.1
56.9
97.4
102.4
169 Spectrin, beta, non-erythrocytic 1 272.5
170 Spectrin, alpha, non-erythrocytic 1 283.7
171
Beaded filament structural protein
1, filensin isoform X2
172
Beaded filament structural protein
1, filensin isoform X3
173
Rho GDP dissociation inhibitor
(GDI) beta
174 Glutathione synthetase
175
Tissue specific transplantation antigen P35B
Ectonucleotide
176 pyrophosphatase/phosphodiesteras e 2
177 Opticin
178 Complement factor H-related
179 Enolase 1, (alpha)
180 Calsyntenin 1
181 Agrin
182 Crystallin, beta B3
183 Seizure related 6 homolog
WAP, Kazal, immunoglobulin,
184 Kunitz and NTR domaincontaining protein 2 isoform X2
185
Aminopeptidase puromycin sensitive
186 Keratin 42, type I
187
Ubiquitin-conjugating enzyme
E2,O
Serpin peptidase inhibitor, clade A
188 (alpha-1 antiproteinase, antitrypsin)
189 Drebrin-like
190 Cell adhesion molecule L1-like
74.2
70.0
45.4
52.1
36.0
100.1
53.0
86.1
50.3
124.5
46.1
43.7
130.3
35.7
35.2
47.3
106.2
207.0
24.3
105.9
apoptosis [23]. Influences the
functional competence of
cryopreserved spermatozoa [24]
Glycoprotein/minimizes apoptosis
[25]
Enzyme
Enzyme
Enzyme
Actin binding protein/regulate cell survival by TNF-alpha-induced
survival signalling [17]
Actin crosslinking and molecular scaffold protein
Actin crosslinking and molecular scaffold protein
Intermediate filament
Intermediate filament
GDP dissociation inhibitor protein
Enzyme
Enzyme
Enzyme
Small Leucine-Rich Repeat Protein
Glycoprotein
Enzyme
Transmembrane protein
Proteoglycan
Structural protein
Transmembrane receptor
Globular protein
Enzyme
Cytoskeletal protein
Enzyme
Enzyme/ inhibits apoptosis by
regulating caspase-3 [26]
Actin binding protein
Transmembrane protein

191 Transketolase isoform X2 63.1
192 Filamin B, beta isoform X5
193 Filamin B, beta isoform X6
275.3
233.2
194
Complement component 4A isoform X1
187.6
195 Aldolase A, fructose-bisphosphate 39.4
196
N-terminal Xaa-Pro-Lys Nmethyltransferase 1
69.7
197 Cadherin-related family member 1 90.0
198
Amyloid beta (A4) precursor-like protein 2
199
Rho GDP dissociation inhibitor alpha
200
Ubiquitin-like modifier activating enzyme 1
201 Cytoplasmic protein
202
StbB stable plasmid inheritance protein B
80.9
45.5
117.8
13.6
14.6
203 Y box binding protein 3
204 Tubulin, beta 2B class IIb
205
ATPase, Na+/K+ transporting, alpha 1 polypeptide
206 LPS-assembly protein
Crystal structure of bovine serum
207 albumin in complex with naproxen, chain B
48.5
10.3
67.0
101.6
66.4
Enzyme
Globular protein
Globular protein
Glycoprotein
Enzyme
Enzyme
Transmembrane protein
Serine-type endopeptidase inhibitor
Inhibitor protein
Enzyme
Plasmid stability protein
Transcription factor
Globular protein
Enzyme
Membrane assembly protein
Globular protein
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