AQP3 antibody (C-Term)

Catalog No. ABIN863208

This anti-AQP3 antibody is a Rabbit Polyclonal antibody detecting AQP3 in WB, IHC, IF and ICC. Suitable for Rat. Independently validated for use in Immunofluorescence.

Quick Overview for AQP3 antibody (C-Term) (ABIN863208)

Target: AQP3 (Aquaporin 3 (Gill Blood Group) (AQP3))

Reactivity: Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This AQP3 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC)

Product Details anti-AQP3 Antibody

Binding Specificity: C-Term

Specificity: Detects ~31.5 kDa. May detect larger glycosylated bands ~35-50 kDa.

Cross-Reactivity: Human, Mouse, Rat

Purification: Protein A Purified

Immunogen: Produced against the C-terminal peptide (Sequence N-CHLEQPPPSTEAENVKLAHMKHKEQI) of rat aquaporin 3

Application Details

Application Notes:

• WB (1:2000)
• IHC (1:200)
• ICC/IF (1:400)
• optimal dilutions for assays should be determined by the user.

Comment:

0.5 μg/ml of ABIN863208 was sufficient for detection of aquaporin 3 in 10 μg of rat kidney tissue lysate by colorimetric immunoblot analysis using Goat anti-rabbit IgG:HRP as the secondary antibody.

Restrictions: For Research Use only

Independent Validation (IF)

Validation #102635 (Immunofluorescence)

by: Centre de Recherche en Transplantation et Immunologie, UMR1064, Université de Nantes

No.: #102635

Date: 12/14/2018

Antigen: AQP3

Lot Number: 160308

Method validated: Immunofluorescence

Positive Control:

Membrane staining in compacted morula; membrane staining apical side, on early blastocysts

Negative Control:

Cells not expressing AQP3 mRNA within human embryos (inner cell mass)

No primary control

Notes:

Passed. ABIN863208 specifically labels the targeted antigen in embryos in IF. No signal was detected in sample negative control tissue and the secondary antibody only control.

Validation Images

Immunoflurescence analysis of human embryo using ABIN863208. AQP3 (right) with nuclear counterstaining (left) at indicated stages. In the top panel (morula), the staining is located at the membrane. In the bottom panel (early blastocyst), the staining is located at the membrane, in trophectoderm cells. Scale bar = 47µm.

Full Methods

Primary Antibody: ABIN863208

Secondary Antibody: donkey anti-rabbit antibody (Life Technologies, A21206)

Full Protocol:

• Fixation
• • Fix embryos in one drop of 30µl of 4% paraformaldehyde for 5min at RT.
  • Remove embryos and wash quickly three times with PBS containing 0.1% BSA (in three drops of 30µl).
• Staining:
• • Permeabilize embryo in 300µl IF buffer for 60min at RT.
  • Incubate embryo with the primary rabbit anti-AQP3 antibody (antibodies-online, ABIN863208, 160308) diluted 1:400 in buffer ON at 4°C.
  • Rinse embryo in IF buffer quickly three times.
  • Incubate embryo with secondary donkey anti-rabbit antibody (Life Technologies, A21206) diluted 1:1000 in buffer for 2h at RT.
  • Rinse embryo in PBS-BSA.
  • At this point, the embryos can either be kept in PBS-BSA ON at 4°C or used for imaging directly on ibido µ-Slide.
  • Image acquisition on a microscope using appropriate filters, 20x oil-immersion objective.

Experimental Notes:

Labeling of human embryo with the aquaporin 3 antibody ABIN863208 shows the expected staining pattern membrane-associated outside of the blastocyst. The background staining in our conditions is a bit high and might be improved with more stringent washes or different fixation-permeabilization protocols.

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: PBS, 50 % glycerol, 0.09 % sodium azide, Storage buffer may change when conjugated

Preservative: Sodium azide

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

Storage: -20 °C

Storage Comment: -20°C

Target Details for AQP3

Target: AQP3 (Aquaporin 3 (Gill Blood Group) (AQP3))

Alternative Name: Aquaporin 3

Background: Aquaporins selectively conduct water molecules in and out of the cell, while preventing the passage of ions and other solutes. Known as water channels, they are integral membrane pore proteins (1, 2). Aquaporin 3 is found in the basolateral cell membrane of principal collecting duct cells and provide a pathway for water to exit these cells (3). AQP3 gene expression is not regulated by vasopressin (4).

Gene ID: 65133

NCBI Accession: NP_113891

UniProt: P47862