CA12 antibody (AA 161-318)

Catalog No. ABIN1858202

The Rabbit Polyclonal anti-CA12 antibody has been validated for WB, IHC, IP and ICC and has been independently validated. It is suitable to detect CA12 in samples from Mouse.

Quick Overview for CA12 antibody (AA 161-318) (ABIN1858202)

Target: CA12 (Carbonic Anhydrase 12 (CA12))

Reactivity: Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This CA12 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Immunocytochemistry (ICC)

Product Details anti-CA12 Antibody

Binding Specificity: AA 161-318

Purpose: Polyclonal Antibody to Carbonic Anhydrase XII (CA12)

Specificity: The antibody is a rabbit polyclonal antibody raised against CA12. It has been selected for its ability to recognize CA12 in immunohistochemical staining and western blotting.

Cross-Reactivity: Human, Rat

Purification: Antigen-specific affinity chromatography followed by Protein A affinity chromatography

Immunogen: Recombinant Carbonic Anhydrase XII (CA12)

Isotype: IgG

Application Details

Application Notes: Western blotting: 0.01-2 μg/mL,Immunohistochemistry: 5-20 μg/mL,Immunocytochemistry: 5-20 μg/mL,Optimal working dilutions must be determined by end user.

Comment:

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #100909 (Immunohistochemistry)

by: School of Clinical Sciences, University of Bristol

No.: #100909

Date: 02/21/2017

Antigen: CA12

Lot Number: A20170109527

Method validated: Immunohistochemistry

Positive Control: mouse kidney, mouse ciliary body

Negative Control: secondary antibody only

Validation Images

Immunostaining of murine kidney tissue (A.) and ciliary body sections (B.) with the CA12 antibody ABIN3204699 (green; counterstain with DAPI, blue) as described in the protocol. No staining of the plasma membrane is observed in ciliary body section stains with the secondary antibody alone (C.).

Full Methods

Primary Antibody: ABIN3204699

Secondary Antibody: goat anti-rabbit IgG Alexa Fluor 488 antibody (Life Technology, A-11006, lot 1348202)

Full Protocol:

• Cryo-section:
• • Wash mouse eye and kidney tissue in PBS.
  • Cryo-protection. Suspend tissue in Optimum Cutting Temperature (OCT) embedding media (Thermo Scientific, Lot 03801137).
  • Gently freeze suspended tissue in liquid nitrogen for 5min.
  • Sections performed on a cryostat (LEICA, CM3050S) set to 12µm.
  • Fix sections in 4% paraformaldehyde (PFA) for 5min at room temperature.
  • Rinse sections in PBS 3 times for 5min.
  • Block sections with 5% normal goat serum for 1h at RT.
• Immunostaining:
• • Incubate sections with primary rabbit-anti CA12 antibody (antibodies-online, ABIN3204699, lot A20170109527) diluted 1:100 in PBSA, overnight at 4°C. Include a negative control without primary antibody and a kidney positive control.
  • Rinse sections with PBS 3 times for 5min. Keep negative control in a separate jar.
  • Incubate sections with secondary goat anti-rabbit IgG Alexa Fluor 488 antibody conjugate (Life Technology, A-11006, lot 1348202) diluted 1:500 in 1% Bovine Serum Albumin in PBS (PBSA) buffer for 2h at RT.
  • Counterstain with DAPI in the mounting medium step.
• Wash and mount:
• • Rinse sections in PBS 3 times for 5min.
  • Mount sections in Vectorshield mounting medium with DAPI (Vector laboratories, H-1200, lot ZC0814).
• Microscopy:
• • Image acquisition on an EVOS FL Color fluorescence and transmitted light microscope (Life technologies, Cat no. AMEFC4300) at 20x magnification.

Handling

Format: Liquid

Concentration: 0.5 mg/mL

Buffer: PBS, pH 7.4, containing 0.02 % Sodium azide, 50 % glycerol.

Preservative: Sodium azide

Precaution of Use: WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.

Handling Advice: Avoid repeated freeze-thaw cycles.

Storage: 4 °C,-20 °C

Storage Comment: Store at 4°C for frequent use. Stored at -20°C in a manual defrost freezer for two year without detectable loss of activity. Avoid repeated freeze-thaw cycles.

Expiry Date: 24 months

Target Details for CA12

Target: CA12 (Carbonic Anhydrase 12 (CA12))

Alternative Name: CA12

Background: CA-XII, CAXII, Carbonate dehydratase XII, Tumor antigen HOM-RCC-3.1.3