Calnexin Antibody - #AF5362

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产品: Calnexin 抗体
货号: AF5362
描述: Rabbit polyclonal antibody to Calnexin
应用: WB IHC IF/ICC
反应: Human, Mouse, Rat, Monkey
预测: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
分子量: 90 kDa; 68kD(Calculated).
蛋白号: P27824
RRID: AB_2837847

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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:
Pig(100%), Zebrafish(80%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(80%), Xenopus(90%)
克隆:
Polyclonal
特异性:
Calnexin Antibody detects endogenous levels of total Calnexin.
RRID:
AB_2837847
引用格式: Affinity Biosciences Cat# AF5362, RRID:AB_2837847.
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

Calnexin; CALX_HUMAN; CANX; CNX; FLJ26570; Histocompatibility complex class I antigen binding protein p88; IP90; Major histocompatibility complex class I antigen-binding protein p88; p90;

抗原和靶标

免疫原:
Uniprot:

P27824(CALX_HUMAN) >>Visit HPA database.

基因/基因ID:
CANX(821)
描述:
Calcium-binding protein that interacts with newly synthesized glycoproteins in the endoplasmic reticulum. It may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits. It seems to play a major role in the quality control apparatus of the ER by the retention of incorrectly folded proteins.
序列:
MEGKWLLCMLLVLGTAIVEAHDGHDDDVIDIEDDLDDVIEEVEDSKPDTTAPPSSPKVTYKAPVPTGEVYFADSFDRGTLSGWILSKAKKDDTDDEIAKYDGKWEVEEMKESKLPGDKGLVLMSRAKHHAISAKLNKPFLFDTKPLIVQYEVNFQNGIECGGAYVKLLSKTPELNLDQFHDKTPYTIMFGPDKCGEDYKLHFIFRHKNPKTGIYEEKHAKRPDADLKTYFTDKKTHLYTLILNPDNSFEILVDQSVVNSGNLLNDMTPPVNPSREIEDPEDRKPEDWDERPKIPDPEAVKPDDWDEDAPAKIPDEEATKPEGWLDDEPEYVPDPDAEKPEDWDEDMDGEWEAPQIANPRCESAPGCGVWQRPVIDNPNYKGKWKPPMIDNPSYQGIWKPRKIPNPDFFEDLEPFRMTPFSAIGLELWSMTSDIFFDNFIICADRRIVDDWANDGWGLKKAADGAAEPGVVGQMIEAAEERPWLWVVYILTVALPVFLVILFCCSGKKQTSGMEYKKTDAPQPDVKEEEEEKEEEKDKGDEEEEGEEKLEEKQKSDAEEDGGTVSQEEEDRKPKAEEDEILNRSPRNRKPRRE

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
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Xenopus
90
Zebrafish
80
Chicken
80
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

翻译修饰 - P27824 作为底物

Site PTM Type Enzyme
T59 O-Glycosylation
K61 Ubiquitination
T66 O-Glycosylation
Y70 Phosphorylation
S74 Phosphorylation
R77 Methylation
T79 O-Glycosylation
S81 Phosphorylation
S86 Phosphorylation
K87 Ubiquitination
T93 Phosphorylation
K99 Ubiquitination
K103 Acetylation
K103 Ubiquitination
K110 Ubiquitination
K118 Ubiquitination
R125 Methylation
K127 Ubiquitination
K134 Acetylation
K134 Ubiquitination
K137 Acetylation
K137 Ubiquitination
K170 Ubiquitination
K182 Ubiquitination
Y185 Phosphorylation
K193 Ubiquitination
K199 Acetylation
K199 Ubiquitination
K210 Ubiquitination
T211 Phosphorylation
Y214 Phosphorylation
K217 Acetylation
K217 Ubiquitination
K227 Acetylation
K227 Ubiquitination
T228 Phosphorylation
K233 Ubiquitination
R282 Methylation
K283 Ubiquitination
K300 Ubiquitination
S362 Phosphorylation
Y379 Phosphorylation
K380 Ubiquitination
Y393 Phosphorylation
K398 Ubiquitination
K401 Ubiquitination
S431 Phosphorylation
K458 Acetylation
K458 Ubiquitination
K459 Acetylation
K459 Ubiquitination
T509 Phosphorylation
S510 Phosphorylation
Y514 Phosphorylation
K516 Ubiquitination
K525 Ubiquitination
K537 Ubiquitination
K547 Ubiquitination
S554 Phosphorylation
T562 Phosphorylation
S564 Phosphorylation
S583 Phosphorylation P27361 (MAPK3) , P06493 (CDK1)

研究背景

功能:

Calcium-binding protein that interacts with newly synthesized glycoproteins in the endoplasmic reticulum. It may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits. It seems to play a major role in the quality control apparatus of the ER by the retention of incorrectly folded proteins. Associated with partial T-cell antigen receptor complexes that escape the ER of immature thymocytes, it may function as a signaling complex regulating thymocyte maturation. Additionally it may play a role in receptor-mediated endocytosis at the synapse.

翻译修饰:

Phosphorylated at Ser-564 by MAPK3/ERK1. phosphorylation by MAPK3/ERK1 increases its association with ribosomes (By similarity).

Palmitoylation by DHHC6 leads to the preferential localization to the perinuclear rough ER. It mediates the association of calnexin with the ribosome-translocon complex (RTC) which is required for efficient folding of glycosylated proteins.

Ubiquitinated, leading to proteasomal degradation. Probably ubiquitinated by ZNRF4.

细胞定位:

Endoplasmic reticulum membrane>Single-pass type I membrane protein. Endoplasmic reticulum. Melanosome.
Note: Identified by mass spectrometry in melanosome fractions from stage I to stage IV (PubMed:12643545, PubMed:17081065). The palmitoylated form preferentially localizes to the perinuclear rough ER (PubMed:22314232).

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:

Interacts with MAPK3/ERK1 (By similarity). Interacts with KCNH2. Associates with ribosomes (By similarity). Interacts with SGIP1; involved in negative regulation of endocytosis (By similarity). The palmitoylated form interacts with the ribosome-translocon complex component SSR1, promoting efficient folding of glycoproteins. Interacts with SERPINA2P/SERPINA2 and with the S and Z variants of SERPINA1. Interacts with PPIB. Interacts with ZNRF4. Interacts with SMIM22.

(Microbial infection) Interacts with HBV large envelope protein, isoform L.

(Microbial infection) Interacts with HBV large envelope protein, isoform M; this association may be essential for isoform M proper secretion.

蛋白家族:

Belongs to the calreticulin family.

研究领域

· Cellular Processes > Transport and catabolism > Phagosome.   (View pathway)

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

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

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

· Organismal Systems > Endocrine system > Thyroid hormone synthesis.

文献引用

1). Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation. Nature Biomedical Engineering (PubMed: 31844155) [IF=28.1]

Application: WB    Species: Mice    Sample: Tumour cells

Fig. 5 | CNP increases exosome release through HSP–p53–TASP6 signalling pathway. a, Simulated temperature changes at five selected locations. A 200 V and 10 ms pulse created a localized ‘hot spot’ in the nanochannel outlet with a power density of ~1 × 1014 W m−3 and a peak temperature up to 60 °C from room temperature. Once the pulse ended, the hot spot vanished rapidly due to the extremely small volume of the heated fluid inside the nanochannel (~1 × 10−12 cm3 ) compared with the bulk solution outside the nanochannel (~0.1 cm3 ). b, Top-down images of MEFs (green) attaching to the surface of the CNP device. Red dots show nanochannel locations and room temperature before CNP transfection (0 s). White arrows indicate locations of the nanochannels. The CNP electric pulse (CNP) sharply increases temperature at the nanochannel–cell surface interface. c, Cross-section view of nanochannels shows temperature changes in the nanochannels before (0 s), during and after (1 s) a CNP pulse. d, Temperature at the cell–nanochannel interface transiently (<1 s) increases to ~60 °C. e, Western blot of HSP90 and HSP70 from untreated (PBS) and CNP (with PBS)-stimulated (CNP) MEFs. f, DLS measurements of exosome concentrations from 108 CNP-stimulated MEFs with or without HSP inhibitors show that HSP70 and HSP90 are critical for the production of exosomes. NVP-HSP990, HSP90 inhibitor; VER155008, HSP70 inhibitor. g, Western blots show that CNP increases expression of p53 and TSAP6 protein in p53 wild-type MEFs, but does not affect p53 or TSAP6 protein expression in p53−/− MEFs. h, DLS measurements of exosome concentrations show that knockdown of p53 can partially block exosome release after CNP. i, Schematic of a proposed mechanism for CNP triggering of exosome release in CNP-transfected cells. Data are from three independent experiments and are presented as mean ± s.e.m. Two-sided Student’s t-test was used for the comparison.
2). Isothiazolinone dysregulates the pattern of miRNA secretion: Endocrine implications for neurogenesis. Environment international (PubMed: 37939439) [IF=11.8]
3). Small extracellular vesicles encapsulating lefty1 mRNA inhibit hepatic fibrosis. Asian Journal of Pharmaceutical Sciences (PubMed: 36382306) [IF=10.2]
4). OSCC cell-secreted exosomal CMTM6 induced M2-like macrophages polarization via ERK1/2 signaling pathway. Cancer immunology, immunotherapy : CII (PubMed: 33104837) [IF=5.8]

Application: WB    Species: Mice    Sample: M0 cells

Fig. 4 The isolation and identifcation of Cal-27 exosomes. a A sketch of isolation path and identifcation of exosomes. b Western blot analyzed of exosomes for CD63, CD9 and CD81 (exosome biomarker), Calnexin (negative marker) and CMTM6. c Western blot analyzed of M0 cells and exosomes incubated M0 cells for CMTM6. d Representative fuorescent images for PKH26-labeled exosomes taken by M0 after 12 h incubation. Exosomes were stained red and M0 nuclei were stained blue by DAPI (scale bar: 25um). Cal-27 cells incubated with non-labeled exosomes (PBS was added to 25 μg exosomes instead of PKH26) and Cal-27 cells with only PKH26 (PKH26 was added to PBS and was re-isolated as stated) were used as controls
5). Endothelial Dysfunction, Inflammation and Coronary Artery Disease: Potential Biomarkers and Promising Therapeutical Approaches. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (PubMed: 33917744) [IF=5.6]
6). Exosomes derived from smooth muscle cells ameliorate diabetes‐induced erectile dysfunction by inhibiting fibrosis and modulating the NO/cGMP pathway. Journal of Cellular and Molecular Medicine (PubMed: 33009701) [IF=5.3]

Application: WB    Species: Rat    Sample: CCSMCs

FIGURE 1Cell identification and exosome characterization. (A) Representative flow cytometry histograms of BMSCs and ADSCs show positive staining for CD29 and CD90 but not for CD34 and CD45. (B) BMSCs and ADSCs were successfully induced into osteoblasts (positively stained with Alizarin Red S) and adipocytes (positively stained with Oil Red O). The magnification is 100×. (C) Representative immunofluorescence results of CCSMCs show positive expression for α-SMA and desmin. Scale bars = 50 μm. (D) Exosomes derived from CCSMCs, BMSCs and ADSCs were observed using transmission electron microscopy, and the particle size distributions of the exosomes were measured by nanoparticle tracking analysis. Scale bars = 100 nm. (E) Representative results of Western blot analysis of exosomes derived from CCSMCs, BMSCs and ADSCs show positive expression for CD9, CD63 and TSG101 but not for calnexin. CCSMC: corpus cavernosum smooth muscle cell; BMSC: bone marrow stem cell; ADSC: adipose-derived stem cell; CCSMC-EXOs: exosomes derived from corpus cavernosum smooth muscle cells; BMSC-EXOs: exosomes derived from bone marrow stem cells; ADSC-EXOs: exosomes derived from adipose-derived stem cells; α-SMA: α-smooth muscle actin; DAPI: 4’,6-diamidino-2-phenylindole
7). Seminal exosomal miR‐210‐3p as a potential marker of Sertoli cell damage in Varicocele. Andrology (PubMed: 33000559) [IF=4.5]
8). hucMSC-Ex Alleviates IBD-Associated Intestinal Fibrosis by Inhibiting ERK Phosphorylation in Intestinal Fibroblasts. Stem Cells International (PubMed: 36845967) [IF=4.3]
9). Exosomes released from Sertoli cells contribute to the survival of Leydig cells through CCL20 in rats. Molecular Human Reproduction (PubMed: 35088858) [IF=4.0]

Application: WB    Species: Rat    Sample:

Figure 2. Sertoli cell-released exosomes promote the survival of Leydig cells. (A) Identification of exosomes with particle size measurement. Exosomes were isolated from rat primary Sertoli cells and measured with nanoparticle tracking analysis (n = 3). (B) Identification of exosomes with CD81 (flow cytometer) and heat shock protein 70 (HSP70) (western blot). Calnexin (CANX) was detected as a negative control (n = 3). See Supplementary Information for the full immunoblot. (C) Coculture of PKH67 labeled exosomes and Leydig cells. Sertoli cell-released exosomes were labeled with PKH67 and added to the supernatant of Leydig cells. After 24 h, the cells were examined under a fluorescence microscope (n = 3). Representative images are shown. (D) Sertoli cell-released exosomes promoted the survival of Leydig cells. The exosomes were added to the supernatant of Leydig cells, and cell survival was detected with cell counting kit-8 (CCK-8 kit) (OD 450 nm value) after 24 h (n = 3, mean ± SD, Student’s t-test). NC, normal control. *P < 0.05.
10). Schwann cell‑derived exosomes induce bone marrow‑derived mesenchymal stem cells to express Schwann cell markers in vitro. Molecular Medicine Reports (PubMed: 32016464) [IF=3.4]

Application: WB    Species: rat    Sample: fibroblasts and RSC96 cells

Figure 1. |Extraction of exosomes from fibroblasts and RSC96 cells. (A) Transmission electron microscopy images of exosomes (yellow arrows) extracted from fibroblasts or RSC96 Schwann cells (magnification, x25,000). Scale bar, 200 nm. (B) Protein expression of exosome marker proteins CD81 and CD63, and the endoplasmic reticulum marker Calnexin were assessed by western blotting.
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