CLEC14A antibody (AA 260-310)

Catalog No. ABIN1385496

This anti-CLEC14A antibody is a Rabbit Polyclonal antibody detecting CLEC14A in WB, IF (p) and IHC (p). Suitable for Human, Mouse and Rat. Independently validated for use in Immunohistochemistry (Formalin-fixed Paraffin-embedded Sections).

Quick Overview for CLEC14A antibody (AA 260-310) (ABIN1385496)

Target: CLEC14A (C-Type Lectin Domain Family 14, Member A (CLEC14A))

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This CLEC14A antibody is un-conjugated

Application: Western Blotting (WB), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Product Details anti-CLEC14A Antibody

Binding Specificity: AA 260-310

Cross-Reactivity: Human, Mouse, Rat

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human EGFR5/CLEC14A

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
IHC-P 1:200-400
IF(IHC-P) 1:50-200

Restrictions: For Research Use only

Independent Validation (IHC (fp))

Validation #100004 (Immunohistochemistry (Formalin-fixed Paraffin-embedded Sections))

by: Memorial Sloan-Kettering Cancer Center, Parada lab

No.: #100004

Date: 11/23/2015

Antigen: CLEC14A

Method validated: Immunohistochemistry (Formalin-fixed Paraffin-embedded Sections)

Positive Control: Tumor cell transplanted mouse brain was dissected

Validation Images

IF validation of CLEC14A antibody ABIN1385496 on mouse brain tumor sections. A. ABIN1385496 diluted 1:100 (blue), DAPI counterstaining (red). B. Dilution factor 1:100. C. Dilution factor 1:200. D. Dilution factor 1:400.

Full Methods

Primary Antibody: ABIN1385496

Secondary Antibody: Donkey-anti-Rabbit-Cy3

Full Protocol:

• Tumor cell transplanted mouse brain was dissected, fixed in 4% PFA for overnight, saturated in 30% sucrose, mounted in Cryo, and sectioned into 12µm sections.
• Immunostaining for Cryo Sections:
• Dry slides at 50-55°C for 15-30 min (or RT for 30min)
• Wash three times with PBS 5min (shake gently)
• Block with 10% Normal Donkey Serum (NDS) at RM for 1hour
• Incubate with primary antibody at 4°C overnight (Prepared in 2% NDS)
• Rabbit-anti-CLEC14A antibody (ABIN1385496) diluted 1:50-1:400
• Wash three times with PBS 5min (shake gently)
• Incubate with secondary antibody together with DAPI at RT for 1h (Prepared in 2% NDS), cover the chamber with foil
• Donkey-anti-Rabbit-Cy3
• DAPI 1:100
• Wash three times with PBS 5min (shake gently)
• Add three drops of water-based Immu-mount
• Cover with 24x60 cover glass
• Imaging
• Images were taken using a Zeiss Axio2-Imaging system, 20X-0.5NA-Dry Dye-1004-072 objective, DAPI and Cy3 channel.

Experimental Notes: ABIN1385496 worked within the diution range 1:50 - 1:200; the antibody did not work well at a 1:400 dilution.

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

Target Details for CLEC14A

Target: CLEC14A (C-Type Lectin Domain Family 14, Member A (CLEC14A))

Alternative Name: EGFR5

Background:

Synonyms: CEG1, EGFR-5, C14orf27, C-type lectin domain family 14 member A, Epidermal growth factor receptor 5, CLEC14A, EGFR5, UNQ236/PRO269

Background: This gene encodes a member of the C-type lectin/C-type lectin-like domain (CTL/CTLD) superfamily. Members of this family share a common protein fold and have diverse functions, such as cell adhesion, cell-cell signalling, glycoprotein turnover, and roles in inflammation and immune response. This family member plays a role in cell-cell adhesion and angiogenesis. It functions in filopodia formation, cell migration and tube formation. Due to its presence at higher levels in tumor endothelium than in normal tissue endothelium, it is considered to be a candidate for tumor vascular targeting.

Gene ID: 161198

UniProt: Q86T13