Product: Occludin Antibody
Catalog: DF7504
Source: Rabbit
Application: WB, IHC, IF/ICC, ELISA(peptide)
Reactivity: Human, Mouse, Rat, Pig
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
Mol.Wt.: 23~30kD, 52~65kD; 59kD(Calculated).
Uniprot: Q16625
RRID: AB_2841004

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Product Info

Source:
Rabbit
Application:
WB 1:1000-3000, IHC 1:50-1:200, IF/ICC 1:200, ELISA(peptide) 1:20000-1:40000
*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,Pig
Prediction:
Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(80%)
Clonality:
Polyclonal
Specificity:
Occludin Antibody detects endogenous levels of total Occludin.
RRID:
AB_2841004
Cite Format: Affinity Biosciences Cat# DF7504, RRID:AB_2841004.
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

BLCPMG; FLJ08163; FLJ18079; FLJ77961; FLJ94056; MGC34277; Occludin; Ocln; OCLN_HUMAN; Tight junction protein occludin;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q16625 OCLN_HUMAN:

Localized at tight junctions of both epithelial and endothelial cells. Highly expressed in kidney. Not detected in testis.

Sequence:
MSSRPLESPPPYRPDEFKPNHYAPSNDIYGGEMHVRPMLSQPAYSFYPEDEILHFYKWTSPPGVIRILSMLIIVMCIAIFACVASTLAWDRGYGTSLLGGSVGYPYGGSGFGSYGSGYGYGYGYGYGYGGYTDPRAAKGFMLAMAAFCFIAALVIFVTSVIRSEMSRTRRYYLSVIIVSAILGIMVFIATIVYIMGVNPTAQSSGSLYGSQIYALCNQFYTPAATGLYVDQYLYHYCVVDPQEAIAIVLGFMIIVAFALIIFFAVKTRRKMDRYDKSNILWDKEHIYDEQPPNVEEWVKNVSAGTQDVPSPPSDYVERVDSPMAYSSNGKVNDKRFYPESSYKSTPVPEVVQELPLTSPVDDFRQPRYSSGGNFETPSKRAPAKGRAGRSKRTEQDHYETDYTTGGESCDELEEDWIREYPPITSDQQRQLYKRNFDTGLQEYKSLQSELDEINKELSRLDKELDDYREESEEYMAAADEYNRLKQVKGSADYKSKKNHCKQLKSKLSHIKKMVGDYDRQKT

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
Horse
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Chicken
80
Xenopus
67
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q16625 As Substrate

Site PTM Type Enzyme
S8 Phosphorylation
K18 Ubiquitination
S40 Phosphorylation
Y44 Phosphorylation
S45 Phosphorylation
K276 Ubiquitination
S277 Phosphorylation
K283 Ubiquitination
Y287 Phosphorylation
K299 Ubiquitination
S302 Phosphorylation
T305 Phosphorylation
S310 Phosphorylation
S313 Phosphorylation
Y315 Phosphorylation
S321 Phosphorylation
Y325 Phosphorylation
S326 Phosphorylation
S327 Phosphorylation
K330 Ubiquitination
Y337 Phosphorylation
S340 Phosphorylation
S341 Phosphorylation
Y342 Phosphorylation
K343 Ubiquitination
S344 Phosphorylation
T345 Phosphorylation
S358 Phosphorylation
Y368 Phosphorylation
S369 Phosphorylation
S370 Phosphorylation
T376 Phosphorylation
S378 Phosphorylation
K379 Ubiquitination
T393 Phosphorylation
Y398 Phosphorylation P12931 (SRC)
T400 Phosphorylation P68400 (CSNK2A1)
Y402 Phosphorylation P12931 (SRC)
T403 Phosphorylation Q02156 (PRKCE) , P24723 (PRKCH) , Q05513 (PRKCZ)
T404 Phosphorylation Q02156 (PRKCE) , Q05513 (PRKCZ) , P24723 (PRKCH) , P68400 (CSNK2A1) , P67870 (CSNK2B)
S408 Phosphorylation P67870 (CSNK2B) , P68400 (CSNK2A1)
T424 Phosphorylation Q02156 (PRKCE) , Q05513 (PRKCZ)
T438 Phosphorylation Q02156 (PRKCE) , Q05513 (PRKCZ)
Y443 Phosphorylation
S445 Phosphorylation
S448 Phosphorylation
S458 Phosphorylation
Y467 Phosphorylation
S471 Phosphorylation
Y474 Phosphorylation
Y481 Phosphorylation
K488 Ubiquitination
S490 Phosphorylation P05771 (PRKCB)

Research Backgrounds

Function:

May play a role in the formation and regulation of the tight junction (TJ) paracellular permeability barrier. It is able to induce adhesion when expressed in cells lacking tight junctions.

PTMs:

Dephosphorylated by PTPRJ. The tyrosine phosphorylation on Tyr-398 and Tyr-402 reduces its ability to interact with TJP1. Phosphorylation at Ser-490 also attenuates the interaction with TJP1.

Subcellular Location:

Cell membrane>Multi-pass membrane protein. Cell junction>Tight junction.

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

Localized at tight junctions of both epithelial and endothelial cells. Highly expressed in kidney. Not detected in testis.

Subunit Structure:

Interacts with TJP1/ZO1. Interacts with VAPA. Interacts with CLDN1, CLDN6, CLDN9, CLDN11, CLDN12 and CLDN17.

Family&Domains:

The C-terminal is cytoplasmic and is important for interaction with ZO-1. Sufficient for the tight junction localization. Involved in the regulation of the permeability barrier function of the tight junction (By similarity). The first extracellular loop participates in an adhesive interaction.

Belongs to the ELL/occludin family.

Research Fields

· Cellular Processes > Cellular community - eukaryotes > Tight junction.   (View pathway)

· Environmental Information Processing > Signaling molecules and interaction > Cell adhesion molecules (CAMs).   (View pathway)

· Human Diseases > Infectious diseases: Bacterial > Pathogenic Escherichia coli infection.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Organismal Systems > Immune system > Leukocyte transendothelial migration.   (View pathway)

References

1). Li X et al. Multistage-Responsive Nanocomplexes Attenuate Ulcerative Colitis by Improving the Accumulation and Distribution of Oral Nucleic Acid Drugs in the Colon. ACS Appl Mater Interfaces 2022 Jan 12;14(1):2058-2070. (PubMed: 34978415) [IF=10.383]

2). Wang Y et al. Fourteen composite probiotics alleviate type 2 diabetes through modulating gut microbiota and modifying M1/M2 phenotype macrophage in db/ db mice. Pharmacol Res 2020 Nov;161:105150. (PubMed: 32818655) [IF=10.334]

Application: IHC    Species: mouse    Sample: colon

Fig. 4.| Effects of the composite probiotics on intestinal barrier function.The protein expression of claudin-1 (G), occludin-1 (H), ZO-1 (I) and mucin-2 (J)determined by immunohistochemistry (original magnification 200×) (n = 4 images/group).

3). Li C et al. Oxyberberine, a novel gut microbiota-mediated metabolite of berberine, possesses superior anti-colitis effect: Impact on intestinal epithelial barrier, gut microbiota profile and TLR4-MyD88-NF-κB pathway. Pharmacol Res 2020 Feb;152:104603 (PubMed: 31863867) [IF=10.334]

Application: WB    Species: Mice    Sample: colonic tissues

Fig. 4. OBB protected intestinal epithelial barrier by modulating TJs proteins. Effect of OBB on the mRNA levels of mucin-1(A) and mucin-2 (B). (C) Representative Western blotting images of TJs protein, and the relative protein expressions were normalized to β-actin. (D-H) Changes in the relative protein expression levels of ZO-1, ZO-2, occludin, JAM-A, and claudin-1 were measured respectively. Data are shown as mean ± SEM (n = 3). # P < 0.05, ## P < 0.01 vs. Control group, * P < 0.05, ** P < 0.01 vs. DSS group, & P < 0.05, && P < 0.01 vs. BBR group.

4). Liu L et al. Extracellular vesicles of Fusobacterium nucleatum compromise intestinal barrier through targeting RIPK1-mediated cell death pathway. Gut Microbes 2021;13(1):1-20. (PubMed: 33769187) [IF=9.434]

Application: IHC    Species: mouse    Sample: colon

Figure 5. |FnEVs increase gut barrier leakage in experimental colitis models. (a) Representative images and ex vivo imaging with the intestine, liver, heart, spleen and kidney of mice. (b) Relative fluorescence intensity of translocated EGFP-labeled E.coli in every tissues. (c) Representative images of immunohistochemical stainings of ZO-1, claudin-1 and occludin in the colon on day 3 after colitis induction. Scale bar = 50 um.

5). Xiao H et al. Tremella fuciformis polysaccharides ameliorated ulcerative colitis via inhibiting inflammation and enhancing intestinal epithelial barrier function. Int J Biol Macromol 2021 Mar 17;180:633-642. (PubMed: 33744251) [IF=8.025]

Application: IF/ICC    Species: mouse    Sample: colon

Fig. 3. |Effect of TFP on intestinal barrier in DSS-induced colitis mice. (A) Representative TEM images in colon tissues (white arrows indicate TJs, black arrows indicate intestinal villi loss).(B) The mRNA level of TJP1 in colon tissues. (C) The mRNA level of OCLN in colon tissues. (D) The fluorescence intensity of ZO-1. (E) The fluorescence intensity of occludin.(F) Representative fluorescent images of ZO-1 and occludin in colon tissues. All data were shown as mean ± SEM. (n = 6 for each group). ⁎P < 0.05. ⁎⁎P < 0.01 compared with control group; #P < 0.05. ##P < 0.01 compared with DSS group.

6). Deng Z et al. Gly-Pro-Ala peptide and FGSHF3 exert protective effects in DON-induced toxicity and intestinal damage via decreasing oxidative stress. Food Res Int 2021 Jan;139:109840. (PubMed: 33509464) [IF=7.425]

7). Wu J et al. Patchouli alcohol attenuates 5-fluorouracil-induced intestinal mucositis via TLR2/MyD88/NF-kB pathway and regulation of microbiota. Biomed Pharmacother 2020 Jan 28;124:109883 (PubMed: 32004938) [IF=7.419]

Application: WB    Species: rat    Sample: Intestinal

Fig. 4. |Effect of PA on intestinal mocusal barrier proteins. (a–d) Relative mRNA expression of MLCK, ZO-1, occludin and claudin-1 (n = 4); (e–i) Expressions of MLC,p-MLC, ZO-1, occludin and claudin-1proteins (n = 3). Values were represented the mean ± SEM. **P < 0.01, *P < 0.05 versus 5-FU group and ##P < 0.01 versus normal group.

8). Zhan X et al. Polysaccharides from Garlic Protect against Liver Injury in DSS-Induced Inflammatory Bowel Disease of Mice via Suppressing Pyroptosis and Oxidative Damage. Oxid Med Cell Longev 2022 Aug 16;2022:2042163. (PubMed: 36017235) [IF=7.310]

9). Tang W et al. The lncRNA-AK046375 Upregulates Metallothionein-2 by Sequestering miR-491-5p to Relieve the Brain Oxidative Stress Burden after Traumatic Brain Injury. Oxid Med Cell Longev 2022 Feb 16;2022:8188404. (PubMed: 35222805) [IF=7.310]

Application: WB    Species: Mice    Sample:

Figure 7 AK046375 maintains BBB integrity and decreases brain water content in mice on 7 days after TBI. (a) Western blotting results of ZO1, occludin, and claudin-5 in each group and quantification (n = 6/group, mean ± SD). (b) Brain water content in each group after TBI (n = 6/group, mean ± SD). (c, d) Fluorescence of Evans blue and Evans blue extravasation in each group around the injury site (n = 12/group, mean ± SD) (scale bar = 50 μm, 200x) (∗P < 0.05 vs. the overexpression vector group, &P < 0.05 vs. the knockdown vector group by one-way ANOVA).

10). Lin Y et al. Hypoxia activates SUMO-1-HIF-1α signaling pathway to upregulate pro-inflammatory cytokines and permeability in human tonsil epithelial cells. Life Sci 2021 Mar 29;276:119432. (PubMed: 33794253) [IF=6.780]

Application: IHC    Species: Human    Sample: HTEC cells

Fig. 3. SUMO1 silencing contributes to reduction in permeability of HTEC cells under hypoxic condition. (A). Knockdown of SUMO1 inhibited the secretion of IL-6, IL-8 and TNF-α in HTEC cells treated under hypoxic condition for 24 h. ***p < 0.001 vs control, !!p < 0.01 vs siNC. (B). SUMO1 silencing promoted the TEER value in HTEC cells treated under hypoxic condition for 24 h. ***p < 0.001 vs control, !!!p < 0.001 vs siNC. (C). SUMO-1 silencing reduced the permeability of HTEC cells treated under hypoxic condition for 24 h. *p < 0.05 vs control, ***p < 0.001 vs siNC; !!!p < 0.001 vs siNC. (D). Western blotting was used to examine the protein levels of SUMO1, HIF-1α, VEGF, Occluding, Claudin-1 and ZO-1 in cells as indicated. (E). Immunohistochemistry staining assay was performed to determine the levels of ZO-1, Occluding and Claudin-1 in cells as indicated above.

Application: WB    Species: Human    Sample: HTEC cells

Fig. 3. SUMO1 silencing contributes to reduction in permeability of HTEC cells under hypoxic condition. (A). Knockdown of SUMO1 inhibited the secretion of IL-6, IL-8 and TNF-α in HTEC cells treated under hypoxic condition for 24 h. ***p < 0.001 vs control, !!p < 0.01 vs siNC. (B). SUMO1 silencing promoted the TEER value in HTEC cells treated under hypoxic condition for 24 h. ***p < 0.001 vs control, !!!p < 0.001 vs siNC. (C). SUMO-1 silencing reduced the permeability of HTEC cells treated under hypoxic condition for 24 h. *p < 0.05 vs control, ***p < 0.001 vs siNC; !!!p < 0.001 vs siNC. (D). Western blotting was used to examine the protein levels of SUMO1, HIF-1α, VEGF, Occluding, Claudin-1 and ZO-1 in cells as indicated. (E). Immunohistochemistry staining assay was performed to determine the levels of ZO-1, Occluding and Claudin-1 in cells as indicated above.

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