GRP78 Antibody - #AF5366

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产品: GRP78 抗体
货号: AF5366
描述: Rabbit polyclonal antibody to GRP78
应用: WB IHC IF/ICC
反应: Human, Mouse, Rat, Monkey
预测: Rabbit, Chicken
分子量: 60~100 kDa; 72kD(Calculated).
蛋白号: P11021
RRID: AB_2837851

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产品描述

来源:
Rabbit
应用:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: 适用于变性蛋白样本的免疫印迹检测. IHC: 适用于组织样本的石蜡(IHC-p)或冰冻(IHC-f)切片样本的免疫组化/荧光检测. IF/ICC: 适用于细胞样本的荧光检测. ELISA(peptide): 适用于抗原肽的ELISA检测.

反应:
Human,Mouse,Rat,Monkey
预测:
Rabbit(100%), Chicken(82%)
克隆:
Polyclonal
特异性:
GRP78 Antibody detects endogenous levels of total GRP78.
RRID:
AB_2837851
引用格式: Affinity Biosciences Cat# AF5366, RRID:AB_2837851.
偶联:
Unconjugated.
纯化:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
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.
别名:

展开/折叠

78 kDa glucose regulated protein; 78 kDa glucose-regulated protein; AL022860; AU019543; BIP; D2Wsu141e; D2Wsu17e; Endoplasmic reticulum lumenal Ca(2+)-binding protein grp78; Endoplasmic reticulum lumenal Ca2+ binding protein grp78; Epididymis secretory sperm binding protein Li 89n; FLJ26106; Glucose Regulated Protein 78kDa; GRP 78; GRP-78; GRP78; GRP78_HUMAN; Heat shock 70 kDa protein 5; Heat Shock 70kDa Protein 5; Heat shock protein family A (Hsp70) member 5; HEL S 89n; Hsce70; HSPA 5; HSPA5; Immunoglobulin Heavy Chain Binding Protein; Immunoglobulin heavy chain-binding protein; mBiP; MIF2; Sez7;

抗原和靶标

免疫原:
Uniprot:

P11021(BIP_HUMAN) >>Visit HPA database.

基因/基因ID:
HSPA5(3309)
描述:
GRP78 a member of the HSP family of molecular chaperones required for endoplasmic reticulum integrity and stress-induced autophagy. Plays a central role in regulating the unfolded protein response (UPR), and is an obligatory component of autophagy in mammalian cells.. May play an important role in cellular adaptation and oncogenic survival. One of the client proteins of GRP78 is protein double-stranded RNA-activated protein-like endoplasmic reticulum kinase (PERK).
序列:
MKLSLVAAMLLLLSAARAEEEDKKEDVGTVVGIDLGTTYSCVGVFKNGRVEIIANDQGNRITPSYVAFTPEGERLIGDAAKNQLTSNPENTVFDAKRLIGRTWNDPSVQQDIKFLPFKVVEKKTKPYIQVDIGGGQTKTFAPEEISAMVLTKMKETAEAYLGKKVTHAVVTVPAYFNDAQRQATKDAGTIAGLNVMRIINEPTAAAIAYGLDKREGEKNILVFDLGGGTFDVSLLTIDNGVFEVVATNGDTHLGGEDFDQRVMEHFIKLYKKKTGKDVRKDNRAVQKLRREVEKAKRALSSQHQARIEIESFYEGEDFSETLTRAKFEELNMDLFRSTMKPVQKVLEDSDLKKSDIDEIVLVGGSTRIPKIQQLVKEFFNGKEPSRGINPDEAVAYGAAVQAGVLSGDQDTGDLVLLDVCPLTLGIETVGGVMTKLIPRNTVVPTKKSQIFSTASDNQPTVTIKVYEGERPLTKDNHLLGTFDLTGIPPAPRGVPQIEVTFEIDVNGILRVTAEDKGTGNKNKITITNDQNRLTPEEIERMVNDAEKFAEEDKKLKERIDTRNELESYAYSLKNQIGDKEKLGGKLSSEDKETMEKAVEEKIEWLESHQDADIEDFKAKKKELEEIVQPIISKLYGSAGPPPTGEEDTAEKDEL

种属预测

种属预测:

score>80的预测可信度较高，可尝试用于WB检测。*预测模型主要基于免疫原序列比对，结果仅作参考，不作为质保凭据。

Species
Results
Score
Rabbit
100
Chicken
82
Bovine
64
Dog
64
Zebrafish
50
Xenopus
44
Pig
0
Horse
0
Sheep
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

翻译修饰 - P11021 作为底物

Site PTM Type Enzyme
S4 Phosphorylation
S14 Phosphorylation
T37 Phosphorylation
Y39 Phosphorylation
R60 Methylation
S64 Phosphorylation
Y65 Phosphorylation
T69 O-Glycosylation
T69 Phosphorylation
K81 Ubiquitination
T85 Phosphorylation
S86 Phosphorylation
T91 Phosphorylation
K96 Acetylation
K96 Ubiquitination
S107 Phosphorylation
K113 Acetylation
K113 Ubiquitination
K118 Acetylation
K118 Ubiquitination
K122 Acetylation
K123 Acetylation
K123 Ubiquitination
K125 Acetylation
K125 Ubiquitination
Y127 Phosphorylation
K138 Acetylation
K138 Ubiquitination
T139 Phosphorylation
S146 Phosphorylation
K152 Acetylation
K152 Ubiquitination
K154 Acetylation
K154 Ubiquitination
Y160 Phosphorylation
K163 Acetylation
K163 Ubiquitination
K164 Ubiquitination
Y175 Phosphorylation
K185 Ubiquitination
T189 Phosphorylation
R197 Methylation
T203 O-Glycosylation
T203 Phosphorylation
Y209 Phosphorylation
K213 Ubiquitination
T229 Phosphorylation P11021 (HSPA5)
K268 Acetylation
K268 Methylation
K268 Ubiquitination
S311 Phosphorylation
Y313 Phosphorylation
S319 Phosphorylation
K326 Acetylation
K326 Ubiquitination
K340 Acetylation
K340 Ubiquitination
K344 Ubiquitination
K352 Acetylation
K352 Sumoylation
K352 Ubiquitination
K353 Acetylation
K353 Sumoylation
K353 Ubiquitination
S354 Phosphorylation
S365 Phosphorylation
T366 Phosphorylation
K370 Ubiquitination
K376 Acetylation
K376 Ubiquitination
K382 Ubiquitination
T441 Phosphorylation
K447 Ubiquitination
S448 Phosphorylation
S452 Phosphorylation
T453 Phosphorylation
S455 Phosphorylation
T460 Phosphorylation
K464 Ubiquitination
Y466 Phosphorylation
K474 Ubiquitination
T485 Phosphorylation
R492 Methylation
T518 Phosphorylation
K523 Ubiquitination
T525 Phosphorylation
T527 Phosphorylation
K547 Acetylation
K547 Ubiquitination
S567 Phosphorylation
Y568 Phosphorylation
Y570 Phosphorylation
S571 Phosphorylation
K573 Ubiquitination
K579 Acetylation
K581 Methylation
K585 Acetylation
K585 Methylation
K585 Ubiquitination
S587 Phosphorylation
S588 Phosphorylation
K591 Methylation
K596 Methylation
K601 Acetylation
K601 Ubiquitination
S607 Phosphorylation
K617 Acetylation
K617 Ubiquitination
K619 Acetylation
S632 Phosphorylation
K633 Acetylation
K633 Ubiquitination
Y635 Phosphorylation
S637 Phosphorylation
T643 O-Glycosylation
T643 Phosphorylation
T648 Phosphorylation
K651 Acetylation

翻译修饰 - P11021 作为激酶

Substrate Site Source
P11021 (HSPA5) T229 Uniprot

研究背景

功能:

Endoplasmic reticulum chaperone that plays a key role in protein folding and quality control in the endoplasmic reticulum lumen. Involved in the correct folding of proteins and degradation of misfolded proteins via its interaction with DNAJC10/ERdj5, probably to facilitate the release of DNAJC10/ERdj5 from its substrate (By similarity). Acts as a key repressor of the ERN1/IRE1-mediated unfolded protein response (UPR). In the unstressed endoplasmic reticulum, recruited by DNAJB9/ERdj4 to the luminal region of ERN1/IRE1, leading to disrupt the dimerization of ERN1/IRE1, thereby inactivating ERN1/IRE1 (By similarity). Accumulation of misfolded protein in the endoplasmic reticulum causes release of HSPA5/BiP from ERN1/IRE1, allowing homodimerization and subsequent activation of ERN1/IRE1 (By similarity). Plays an auxiliary role in post-translational transport of small presecretory proteins across endoplasmic reticulum (ER). May function as an allosteric modulator for SEC61 channel-forming translocon complex, likely cooperating with SEC62 to enable the productive insertion of these precursors into SEC61 channel. Appears to specifically regulate translocation of precursors having inhibitory residues in their mature region that weaken channel gating.

翻译修饰:

AMPylated by FICD. In unstressed cells, AMPylation at Thr-518 by FICD inactivates the chaperome activity: AMPylated form is locked in a relatively inert state and only weakly stimulated by J domain-containing proteins (By similarity). In response to endoplasmic reticulum stress, de-AMPylation by the same protein, FICD, restores the chaperone activity (By similarity).

细胞定位:

Endoplasmic reticulum lumen. Melanosome. Cytoplasm.
Note: Identified by mass spectrometry in melanosome fractions from stage I to stage IV.

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

Monomer and homooligomer; homooligomerization via the interdomain linker inactivates the chaperone activity and acts as a storage of HSPA5/BiP molecules (By similarity). Interacts with DNAJC1 (via J domain) (By similarity). Component of an EIF2 complex at least composed of CELF1/CUGBP1, CALR, CALR3, EIF2S1, EIF2S2, HSP90B1 and HSPA5 (By similarity). Part of a large chaperone multiprotein complex comprising DNAJB11, HSP90B1, HSPA5, HYOU, PDIA2, PDIA4, PDIA6, PPIB, SDF2L1, UGT1A1 and very small amounts of ERP29, but not, or at very low levels, CALR nor CANX (By similarity). Interacts with TMEM132A and TRIM21. May form a complex with ERLEC1, OS9, SEL1L and SYVN1. Interacts with DNAJC10. Interacts with DNAJB9/ERdj4; leading to recruit HSPA5/BiP to ERN1/IRE1 (By similarity). Interacts with ERN1/IRE1; interaction takes place following interaction with DNAJB9/ERdj4 and leads to inactivate ERN1/IRE1 (By similarity). Interacts with MX1 (By similarity). Interacts with METTL23. Interacts with CEMIP; the interaction induces calcium leakage from the endoplasmic reticulum and cell migration. Interacts with PCSK4 form; the interaction takes place in the endoplasmic reticulum. Interacts with CIPC. Interacts with CCDC88B (via C-terminus); the interaction opposes ERN1-mediated JNK activation, protecting against apoptosis. Interacts with INPP5K; necessary for INPP5K localization at the endoplasmic reticulum. Interacts with MANF; the interaction is direct. Interacts with LOXL2; leading to activate the ERN1/IRE1-XBP1 pathway of the unfolded protein response. Interacts with CLU under stressed condition; interaction increases CLU protein stability; facilitates its retrotranslocation and redistribution to the mitochondria; cooperatively suppress stress-induced apoptosis by stabilizing mitochondrial membrane integrity. Interacts with CCDC47 (By similarity).

蛋白家族:

The interdomain linker regulates the chaperone activity by mediating the formation of homooligomers. Homooligomers are formed by engagement of the interdomain linker of one HSPA5/BiP molecule as a typical substrate of an adjacent HSPA5/BiP molecule. HSPA5/BiP oligomerization inactivates participating HSPA5/BiP protomers. HSPA5/BiP oligomers probably act as reservoirs to store HSPA5/BiP molecules when they are not needed by the cell. When the levels of unfolded proteins rise, cells can rapidly break up these oligomers to make active monomers.

Belongs to the heat shock protein 70 family.

研究领域

· Genetic Information Processing > Folding, sorting and degradation > Protein export.

· Genetic Information Processing > Folding, sorting and degradation > Protein processing in endoplasmic reticulum.   (View pathway)

· Human Diseases > Neurodegenerative diseases > Prion diseases.

· Organismal Systems > Immune system > Antigen processing and presentation.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone synthesis.

文献引用

1). TRAIL‐Armed ER Nanosomes Induce Drastically Enhanced Apoptosis in Resistant Tumor in Combination with the Antagonist of IAPs (AZD5582). Advanced Healthcare Materials, 2021 (PubMed: 33963815) [IF=10.0]
2). Epimedin B exerts neuroprotective effect against MPTP-induced mouse model of Parkinson's disease: GPER as a potential target. Biomedicine & Pharmacotherapy, 2022 (PubMed: 36411637) [IF=7.5]

Application: WB    Species: Mouse    Sample:

Fig. 7. Epimedin B exerts anti-apoptotic and anti-endoplasmic reticulum stress effects on MPTP-induced PD mice through GPER. Western blot assay was used to assess the effect of Epimedin B on the protein expressions of apoptosis-related protein Bax and Bcl-2 (A), and endoplasmic reticulum stress-related protein GRP78 and CHOP (B). Data are expressed as means ± SD, (n = 6).
3). mTOR pathway mediates endoplasmic reticulum stress-induced CD4+ T cell apoptosis in septic mice. APOPTOSIS, 2022 (PubMed: 35759162) [IF=7.2]

Application: WB    Species: Mice    Sample: CD4+ T cells

Fig. 4 Expression of ERS-UPR- and mTOR-related proteins in splenic CD4+ T cells of septic mice. Protein expression of GRP78, CHOP, mTOR, p-mTOR, p70S6k, p-p70S6k (A–E) in CD4+ T cells were quantified by western blotting and showed as the relative expression values of β-actin, which was used as a loading control to normalize the protein levels. In order to highlight the activation level of p-mTOR and p-p70S6K in this signaling pathway, the ratio of p-mTOR to mTOR (p/t mTOR) and p-p70S6K to p70S6K (p/t p70S6K) were used for statistics. Data are shown as Mean ± SD (n = 6). Statistically significant differences were determined by two-tailed Student’s t-test. **P < 0.01, ***P < 0.001, ****P < 0.0001
4). Aqueous Extract of Pepino Leaves Ameliorates Palmitic Acid-Induced Hepatocellular Lipotoxicity via Inhibition of Endoplasmic Reticulum Stress and Apoptosis. Antioxidants, 2021 (PubMed: 34204987) [IF=7.0]

Application: WB    Species: Human    Sample: HepG2 Cells

Figure 5 AEPL attenuated ER stress while PA exposure. (a) ER stress-related protein levels were analyzed by Western blotting. (b–d) GRP78, phosphorylated PERK, and phosphorylated IRE1αwere normalized to PERK, IRE1α, and β-actin of each sample. (e) SREBP-1 level in each group was normalized to β-actin. All data were presented as mean ± SD of at least three independent experiments. * p < 0.05 vs. control cells; # p < 0.05 vs. PA-treated cells.
5). The role of vitamin E in polyunsaturated fatty acid synthesis and alleviating endoplasmic reticulum stress in sub-adult grass carp (Ctenopharyngodon idella). Animal nutrition (Zhongguo xu mu shou yi xue hui), 2024 (PubMed: 38371478) [IF=6.3]

Application: WB    Species: fish    Sample:

Fig. 5 Effects of vitamin E on the relative protein expressions of endoplasmic reticulum stress-related proteins in the muscle of sub-adult grass carp. (A) Western blots for p-PERK (Ser1096), p-IRE1 (Ser724), ATF6 and GRP78. Quantification of (B) p-PERK (Ser1096), (C) p-IRE1 (Ser724), (D) ATF6, and (E) GRP78 normalized to β-actin. Results are represented as the mean ± SD. The data are the means of three replicates with two fishes per replicate (n = 3). Mean values within the same row with different superscripts are significantly different (P < 0.05). p-PERK (Ser1096) = protein kinase R-like endoplasmic reticulum kinase (phospho-Ser1096); p-IRE1 (Ser724) = inositol-requiring enzyme 1 (phospho-Ser724); ATF6 = activating transcription factor 6; GRP78 = glucose regulatory protein 78.
6). The role of SUMO specific peptidase 3 in secondary inflammation of ischemic stroke in mice. Biochimica et biophysica acta. Molecular basis of disease, 2024 (PubMed: 38437993) [IF=6.2]
7). Sesamol supplementation alleviates nonalcoholic steatohepatitis and atherosclerosis in high-fat, high carbohydrate and high-cholesterol diet-fed rats. Food & Function, 2021 (PubMed: 34606548) [IF=6.1]

Application: WB    Species: Rat    Sample:

Fig. 5 Sesamol regulated the hepatic ERS-IRE1 signaling pathway and NLRP3 expression in HF-HCC diet-fed rats. (A) Representative images of the western blot analyses from hepatic Bip, IRE1, phospho-IRE1, and NLRP3 expression. (B) The semi-quantitative analysis of Bip, and (C and D) representative images of the IHC analyses from hepatic phospho-IRE1 and NLRP3 expression (samples from 3 rats in each group were analyzed). Values are expressed as means ± SEM (n = 3); ###P < 0.001, versus control; ***P < 0.001, versus HF-HCC.
8). GnRHa protects the ovarian reserve by reducing endoplasmic reticulum stress during cyclophosphamide-based chemotherapy. npj Breast Cancer, 2021 (PubMed: 34620881) [IF=5.9]

Application: WB    Species: Human    Sample: KGN cells

Fig. 3 Cyclophosphamide inhibits AMH secretion in vitro. a Representative western blot analysis of GRP78 and AMH from KGN cells at 36 h and 48 h after injection of vehicle or CTX at doses of 250–1250 µg/mL. b Apoptosis assay of KGN cells 36 h and 48 h after injection of vehicle or CTX at doses of 250–1250 µg/mL. Compared with untreated control cells, the level of apoptosis was significantly higher with ≥750 µg/mL CTX. c, d AMH mRNA levels of KGN cells verified by qPCR. e, f The concentration of AMH in the cell-culture medium declined as the CTX dose increased. g, h One-way ANOVA analysis of apoptotic cells displayed on b. All error bars represent the standard error of the mean. *P < 0.05, **P < 0.001, and ***P < 0.0001.
9). The potential immunotoxicity of fine particulate matter based on SD rat spleen. ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH, 2019 (PubMed: 31218585) [IF=5.8]

Application: WB    Species: rat    Sample: spleen

Fig. 3 | PM2.5 induced ERS in spleen of SD rats. a, b The levels of spleen GRP78 and ATF6 mRNA were measured using qRT-PCR by summer and winter PM2.5 treatment in SD rat. c, dThe levels of spleen GRP78 and ATF6 protein were measured using western blotting in summer PM2.5 treatment in SD rats and quantification of analysis
10). Protective Effect of Patchouli Alcohol Against High-Fat Diet Induced Hepatic Steatosis by Alleviating Endoplasmic Reticulum Stress and Regulating VLDL Metabolism in Rats. Frontiers in Pharmacology, 2019 (PubMed: 31632274) [IF=5.6]

Application: WB    Species: rat    Sample: liver

FIGURE 4 | PA treatment attenuated HFD-induced ER stress in rats. (A) Representative immunoreactive bands of GRP78, PERK, p-PERK, IRE1α, p-IRE1α, and ATF6
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