Cadherin antibody (AA 841-882)

Catalog No. ABIN687682

Product Details anti-Cadherin Antibody

Target: Cadherin

Binding Specificity: AA 841-882

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)

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

Abstract: Cadherin Products

Synonyms: ci-cadherin antibody, cadherin antibody, ci-cadherin antibody, LOC430062 antibody

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