Cadherin antibody (AA 841-882)

Catalog No. ABIN687682

This independently validated Rabbit Polyclonal antibody specifically detects Cadherin in WB, FACS, IF (cc), IF (p), IHC (p), ELISA, IHC (fro) and ICC. It exhibits reactivity toward Human and Mouse and has been mentioned in 2+ publications.

Quick Overview for Cadherin antibody (AA 841-882) (ABIN687682)

Target: Cadherin

Reactivity: Human, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Cadherin antibody is un-conjugated

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

Product Details anti-Cadherin Antibody

Binding Specificity: AA 841-882

Cross-Reactivity: Human, Mouse

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

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human E-cadherin

Isotype: IgG

Application Details

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

Restrictions: For Research Use only

Independent Validation (FACS)

Validation #029742 (Flow Cytometry)

by: Flow Cytometry & Cell Separation Facility, Purdue University

No.: #029742

Date: 06/30/2014

Lot Number: 131101

Method validated: Flow Cytometry

Positive Control: MCF7 cells (high expression)

Negative Control: SH-SY5Y cells (low expression)

Notes: Signal was observed in the positive control cells. There was some non-specific staining observed with the isotype and secondary-only antibody controls which was lower than the signal from the anti-E-cadherin antibody. Low signal was observed in the low expressing (negative control) cells as expected.

Validation Images

Figure 1. Histogram of MCF7 cells stained with anti-E-cadherin (orange), isotype control antibody (green), secondary antibody only (blue) and unstained (red).

Figure 2. Histogram of SH-SY5Y cells stained with anti-E-cadherin (orange), isotype control antibody (green), secondary antibody only (blue) and unstained (red).

Full Methods

Primary Antibody:

• Antigen: Cadherin 1, Type 1, E-Cadherin (Epithelial) (CDH1)
• Catalog number: ABIN687682
• Lot number: 131101

Secondary Antibody:

• Antibody: Goat anti-rabbit IgG-Alexa 647

Full Protocol:

• Positive and negative control cells were washed once with phosphate-buffered saline (PBS) and harvested with a non-enzymatic cell dissociation solution (Cellstripper, Mediatech, Inc).
• Detached cells were washed twice and resuspended in 100 µL 1X PBS containing 0.5% BSA: - unstained cells - secondary antibody alone - 1 µg primary antibody - 1 µg isotype control antibody
• Cells were incubated for 30 min on ice.
• Labeled cells were washed twice in PBS containing 0.5% BSA.
• Cells were resuspended with 1X PBS containing 0.5% BSA + 10% goat serum and incubated for 15 min at room temperature.
• Goat anti-rabbit IgG-Alexa 647 secondary antibody (Jackson Immunoresearch) was added at a 1:500 dilution. The cells were incubated for 30 min in the dark on ice.
• Labeled cells were washed twice in PBS containing 0.5% BSA.
• Propidium Iodide (PI) was added to discern live cells from dead cells.
• Cells were analyzed on a FACSAria III (BD Biosciences) using a red laser (640 nm excitation / 660 nm emission).

Experimental Notes: - The data displayed is gated on PI negative cells.

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-Cadherin antibody (ABIN687682)

Yu, Wang, Tang, Zhang, Shang: "RNA interference-mediated knockdown of Notch-1 inhibits migration and invasion, down-regulates matrix metalloproteinases and suppresses NF-?B signaling pathway in trophoblast cells." in: Acta histochemica, Vol. 116, Issue 5, pp. 911-9, (2014) (PubMed).

Fan, Wang, Zhao, Liu, Chen, Sun, Zheng: "Sonic hedgehog signaling may promote invasion and metastasis of oral squamous cell carcinoma by activating MMP-9 and E-cadherin expression." in: Medical oncology (Northwood, London, England), Vol. 31, Issue 7, pp. 41, (2014) (PubMed).

Target Details for Cadherin

Target: Cadherin

Background:

Synonyms: UVO, CDHE, ECAD, LCAM, Arc-1, CD324, Cadherin-1, CAM 12/8, Epithelial cadherin, E-cadherin, Uvomorulin, CDH1

Background: Cadherins are calcium-dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells, cadherins may thus contribute to the sorting of heterogeneous cell types. CDH1 is involved in mechanisms regulating cell-cell adhesions, mobility and proliferation of epithelial cells. Has a potent invasive suppressor role. It is a ligand for integrin alpha-E/beta-7. E-Cad/CTF2 promotes non-amyloidogenic degradation of Abeta precursors. Has a strong inhibitory effect on APP C99 and C83 production.

Gene ID: 999

UniProt: P12830