Occludin antibody (AA 431-522)

Catalog No. ABIN687337

Product Details anti-Occludin Antibody

Target: Occludin (OCLN)

Binding Specificity: AA 431-522

Reactivity: Human, Mouse, Pig

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Occludin antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Flow Cytometry (FACS), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Cross-Reactivity: Human, Mouse, Pig

Predicted Reactivity: Rat,Dog,Cow,Horse,Guinea Pig

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from mouse Occludin

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IF(IHC-P) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (IF)

Validation #029632 (Immunofluorescence)

by: Confocal Imaging Core, Beth Israel Deaconess Medical Center

No.: #029632

Date: 03/23/2014

Lot Number: 990698W

Method validated: Immunofluorescence

Positive Control: Mouse stomach tissue

Negative Control: Human fat tissue

Notes: Signal was detected in positive control sample and not in negative control sample.

Validation Images

Figure 1: Immunostaining (green) of mouse stomach tissue (A-C) and human fat tissue (D-F) with anti-OCLN (A, D), isotype control antibody (B, E), or with secondary alone (C, F). Tissues were counterstained with DAPI (blue).

Full Methods

Primary Antibody:

• Antibody: Occludin (OCLN)
• Catalog number: ABIN687337
• Lot number: 990698W

Secondary Antibody:

• Antibody: Donkey anti-Rabbit IgG (Heavy & Light Chain) Antibody (Alexa 647)

Full Protocol:

• Paraffin embedded positive and negative control tissue sections were deparaffinized and undergone antigen retrieval using 10 mM Sodium Citrate pH6 in a pressure cooker for 10 min.
• The tissue sections were incubated with 1 mg/mL Sodium borohydride for 10 min at room temperature to block autofluorescent background signal. The sections were then rinsed three times in TBS for 5 min each at RT.
• Tissue sections were blocked in 1 X TBS / 5% normal donkey serum for 60 min at RT.
• Tissue sections were incubated with primary antibody diluted 1:200 in 1X PBS / 5% normal donkey serum overnight at 4°C.
• Tissue sections were rinsed three times in TBS for 5 min each at RT.
• Cells were incubated with secondary antibody diluted 1:200 in 1X PBS / 5% normal donkey serum for 60 min at RT in dark.
• Tissue sections were rinsed three times in TBS for 5 min each at RT.
• Coverslips were mounted on slides with Prolong Gold anti-fade mounting media (Invitrogen)
• IF stained tissue sections were imaged with a Zeiss LSM 510 Meta confocal microscope.

Experimental Notes:

• No challenges notes
• OCLN is recommended to staining of mouse stomach tissue according to this protocol.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-Occludin antibody (ABIN687337)

Sun, Liu, Xue, Wang: "Effects of insulin combined with idebenone on blood-brain barrier permeability in diabetic rats." in: Journal of neuroscience research, Vol. 93, Issue 4, pp. 666-77, (2015) (PubMed).

Zhao, Qin, Han, Wang, Zhang, Liu: "?-Conglycinin reduces the tight junction occludin and ZO-1 expression in IPEC-J2." in: International journal of molecular sciences, Vol. 15, Issue 2, pp. 1915-26, (2014) (PubMed).

Pan, Qin, Zhao, Wang, Liu, Che: "Effects of soybean agglutinin on mechanical barrier function and tight junction protein expression in intestinal epithelial cells from piglets." in: International journal of molecular sciences, Vol. 14, Issue 11, pp. 21689-704, (2013) (PubMed).

Zhao, Qin, Sun, Che, Bao, Zhang: "Effects of soybean agglutinin on intestinal barrier permeability and tight junction protein expression in weaned piglets." in: International journal of molecular sciences, Vol. 12, Issue 12, pp. 8502-12, (2012) (PubMed).

Gu, Xue, Wei, Zhang, Li: "Calcium-activated potassium channel activator down-regulated the expression of tight junction protein in brain tumor model in rats." in: Neuroscience letters, Vol. 493, Issue 3, pp. 140-4, (2011) (PubMed).

Target Details for Occludin

Target: Occludin (OCLN)

Alternative Name: Occludin (OCLN Products)

Synonyms: AI503564 antibody, Ocl antibody, ocln antibody, oclnb antibody, tpmt antibody, OCLN antibody, wu:fd23h10 antibody, wu:fi13c01 antibody, zgc:113992 antibody, zgc:56359 antibody, BLCPMG antibody, occludin antibody, occludin S homeolog antibody, occludin a antibody, Ocln antibody, OCLN antibody, ocln.S antibody, ocln antibody, oclna antibody

Background:

Synonyms: Ocl, AI53564, Occludin, Ocln

Background: May play a role in the formation and regulation of the tight junction (TJ) paracellular permeability barrier.

Gene ID: 18260

UniProt: Q61146

Pathways: Cell-Cell Junction Organization, Hepatitis C