Product: BOK Antibody
Catalog: DF3829
Description: Rabbit polyclonal antibody to BOK
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Chicken, Xenopus
Mol.Wt.: 23 KD; 23kD(Calculated).
Uniprot: Q9UMX3
RRID: AB_2836186

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Product Info

Source:
Rabbit
Application:
IHC 1:50-1:200, WB 1:1000-3000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Chicken(92%), Xenopus(83%)
Clonality:
Polyclonal
Specificity:
BOK Antibody detects endogenous levels of total BOK.
RRID:
AB_2836186
Cite Format: Affinity Biosciences Cat# DF3829, RRID:AB_2836186.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Bcl 2 related ovarian killer protein; Bcl-2-like protein 9; Bcl-2-related ovarian killer protein; BCL2 related ovarian killer; Bcl2-L-9; BCL2L9; BOK; BOK_HUMAN; BOKL; Hbok; MGC4631;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q9UMX3 BOK_HUMAN:

Expressed mainly in oocytes; weak expression in granulosa cells of the developing follicles. In adult human ovaries, expressed in granulosa cells at all follicular stages, but expression in primordial/primary follicles granulosa cell is stronger than in secondary and antral follicles.

Sequence:
MEVLRRSSVFAAEIMDAFDRSPTDKELVAQAKALGREYVHARLLRAGLSWSAPERAAPVPGRLAEVCAVLLRLGDELEMIRPSVYRNVARQLHISLQSEPVVTDAFLAVAGHIFSAGITWGKVVSLYAVAAGLAVDCVRQAQPAMVHALVDCLGEFVRKTLATWLRRRGGWTDVLKCVVSTDPGLRSHWLVAALCSFGRFLKAAFFVLLPER

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Chicken
92
Xenopus
83
Horse
0
Sheep
0
Dog
0
Zebrafish
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9UMX3 As Substrate

Site PTM Type Enzyme
S7 Phosphorylation
S8 Phosphorylation
K25 Ubiquitination
S196 Phosphorylation

Research Backgrounds

Function:

Apoptosis regulator that functions through different apoptotic signaling pathways. Plays a roles as pro-apoptotic protein that positively regulates intrinsic apoptotic process in a BAX- and BAK1-dependent manner or in a BAX- and BAK1-independent manner. In response to endoplasmic reticulum stress promotes mitochondrial apoptosis through downstream BAX/BAK1 activation and positive regulation of PERK-mediated unfolded protein response (By similarity). Activates apoptosis independently of heterodimerization with survival-promoting BCL2 and BCL2L1 through induction of mitochondrial outer membrane permeabilization, in a BAX- and BAK1-independent manner, in response to inhibition of ERAD-proteasome degradation system, resulting in cytochrome c release. In response to DNA damage, mediates intrinsic apoptotic process in a TP53-dependent manner. Plays a role in granulosa cell apoptosis by CASP3 activation. Plays a roles as anti-apoptotic protein during neuronal apoptotic process, by negatively regulating poly ADP-ribose polymerase-dependent cell death through regulation of neuronal calcium homeostasis and mitochondrial bioenergetics in response to NMDA excitation (By similarity). In addition to its role in apoptosis, may regulate trophoblast cell proliferation during the early stages of placental development, by acting on G1/S transition through regulation of CCNE1 expression. May also play a role as an inducer of autophagy by disrupting interaction between MCL1 and BECN1.

Pro-apoptotic molecule exerting its function through the mitochondrial pathway.

PTMs:

Ubiquitinated by AMFR/gp78 E3 ubiquitin ligase complex; mediates degradation by ubiquitin-proteasome pathway in a VCP/p97-dependent manner; prevents from pro-apoptotic activity; promotes degradation of newly synthesized proteins that are not ITPR1 associated.

Subcellular Location:

Mitochondrion membrane>Single-pass membrane protein. Endoplasmic reticulum membrane>Single-pass membrane protein. Mitochondrion inner membrane. Cytoplasm. Nucleus. Mitochondrion. Endoplasmic reticulum. Mitochondrion outer membrane. Early endosome membrane. Recycling endosome membrane. Nucleus outer membrane. Golgi apparatus>cis-Golgi network membrane. Golgi apparatus>trans-Golgi network membrane. Membrane.
Note: Nuclear and cytoplasmic compartments in the early stages of apoptosis and during apoptosis it associates with mitochondria (PubMed:19942931). In healthy cells, associates loosely with the membrane in a hit-and-run mode. The insertion and accumulation on membranes is enhanced through the activity of death signals, resulting in the integration of the membrane-bound protein into the membrane (PubMed:15868100). The transmembrane domain controls subcellular localization; constitutes a tail-anchor. Localizes in early and late endosome upon blocking of apoptosis. Must localize to the mitochondria to induce mitochondrial outer membrane permeabilization and apoptosis (By similarity).

Membrane. Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed mainly in oocytes; weak expression in granulosa cells of the developing follicles. In adult human ovaries, expressed in granulosa cells at all follicular stages, but expression in primordial/primary follicles granulosa cell is stronger than in secondary and antral follicles.

Subunit Structure:

Monomer; positively regulates apoptotic process. Homodimer (By similarity). Heterodimer (By similarity). Oligomer; promoted by apoptotic stimuli and BH3-only proteins; mediates constitutive activation. Interacts (via BH4 domain) with ITPR1; enhances BOK expression and stabilization; limits apoptosis and prevents ubiquitination and then degradation; protects ITPR1 from proteolysis by CASP3 during apoptosis. Interacts with ITPR2 AND ITPR3; binds most strongly to ITPR2, and barely to ITPR3; regulates their expression (By similarity). Interacts with XPO1; translocates to the cytoplasm. Interacts with BNIP3; promotes oligomerization.

Family&Domains:

BH4 domain mediates interaction with ITPR1.

Belongs to the Bcl-2 family.

Research Fields

· Cellular Processes > Cell growth and death > Apoptosis - multiple species.   (View pathway)

References

1). SARS-CoV-2 membrane protein causes the mitochondrial apoptosis and pulmonary edema via targeting BOK. Cell Death & Differentiation, 2022 (PubMed: 35022571) [IF=12.4]

Application: WB    Species: Mouse    Sample: lung tissue

Fig. 3 SARS-CoV-2 M protein inhibited BOK ubiquitination and promoted its translocation and interacted with BOK via endodomain. A, B H292 cells were not transfected (control) or transfected with an empty vector (pcDNA6B) or M-Flag plasmid for 48 h. The protein levels of Bcl-2, BID, BAX, BAK and BOK (A) or M-Flag, Precursor caspase-3 (Pro-CASP 3) and cleaved CASP 3 were analyzed by WB. C After H292 cells were transfected with M-Flag for 48 h, the mitochondrial and cytosolic fractions were isolated and analyzed by WB. D mRNA levels of Bok in H292 cells transfection with M-Flag for 48 h were examined by qPCR. E Cycloheximide (CHX) chase to check the turnover of endogenous BOK, H292 cells either transfected with vector or M-Flag expressing plasmid, were treated with 30 μg/ml CHX and harvested at the indicated time periods for WB analysis (upper panel), and quantification of BOK level. β-actin use as internal control (lower panel). F Lysates from H292 non-transfected (control) or transfected with an empty vector (pcDNA6B) or M-Flag plasmid followed by treatment with 10 μM MG132 for 5 h before harvest and were immunoprecipitated. Ub Ubiquitination. G HEK293T cells were cotransfected with plasmids encoding HA-tagged ubiquitin, Myc-tagged BOK, together with either an empty vector (pcDNA3.1(+)) or plasmid encoding Flag-tagged M protein for 40 h. Cells were treated with 10 μM MG132 for 5 h and lysed. Then the proteins mixture was immunoprecipitated with anti-Myc antibody followed by WB. H Immunoprecipitation using anti-Flag or anti-HA antibodies from lysates of 293 T cells transfected with HA-tagged BOK (24 kDa) alone, or with an empty vector (pcDNA6B), SARS-CoV-2 Flag-tagged N (51 kDa) and M (23/24 kDa). I Exogenous protein interaction was further confirmed in 293 T lysates immunoprecipitated with anti-HA and analyzed by WB with the indicated antibodies. J, K 293 T cells were co-transfected with pcDNA3.1(+) carrying HA-tagged BOK, with a pcDNA6B vector containing Flag-tagged M (full lengh), Flag-tagged truncated variants of M proteins (M-▵transmembrane (TM)1-Flag (22 kDa), M-▵TM1-2-Flag (18/20 kDa), M-▵TM1-3-Flag (14 kDa), M-▵endodomain (ED)-Flag (22 kDa)). Blots of cell lysates (input) or anti-Flag (J) and anti-HA (K) immunoprecipitates were analyzed by Western blotting for HA and Flag. L Confocal microscopy of 293 T cells co-transfected with HA-tagged BOK and Flag-tagged M (above lane) or Flag-tagged M-▵ED (below lane). White arrows represent the colocalization. Scale bars, 10 μm. Data present with mean ± SD for at least three independent experiments. ns non-significant, **p < 0.01. Data were analyzed with one-way ANOVA with LSD’ t test.

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