TRPC5 antibody (AA 827-845) (FITC)

Catalog No. ABIN2485379

This anti-TRPC5 antibody is a Mouse Monoclonal antibody detecting TRPC5 in WB, IHC, IP and AA. Suitable for Human. Independently validated for use in Immunofluorescence.

Quick Overview for TRPC5 antibody (AA 827-845) (FITC) (ABIN2485379)

Target: TRPC5 (Transient Receptor Potential Cation Channel, Subfamily C, Member 5 (TRPC5))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This TRPC5 antibody is conjugated to FITC

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Antibody Array (AA)

Clone: N67-15 (Formerly S67-15)

Product Details anti-TRPC5 Antibody (FITC)

Binding Specificity: AA 827-845

Specificity: Detects ~110 kDa.

Cross-Reactivity: Human, Mouse, Rat

Purification: Protein G Purified

Immunogen: Synthetic peptide amino acids 827-845 of human TrpC5 (also known as short transient receptor potential channel 5, and Htrp5)

Isotype: IgG2b

Application Details

Application Notes:

• WB (1:1000)
• IHC (1:1000)
• ICC/IF (1:100)
• optimal dilutions for assays should be determined by the user.

Comment:

1 μg/ml of ABIN2485379 was sufficient for detection of TrpC5 in 20 μg of rat brain lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.

Restrictions: For Research Use only

Independent Validation (IF)

Validation #103875 (Immunofluorescence)

by: Experimentelle Anästhesiologie, Universitätsklinikum Erlangen

No.: #103875

Date: 04/10/2019

Antigen: TRPC5

Lot Number: 1007

Method validated: Immunofluorescence

Positive Control:

C57Bl6/J Mouse WT Trigeminal Ganglion (TG)

Negative Control:

Mouse TRPC5 knock-out Trigeminal Ganglion

Notes:

ABIN2485379 specifically stains TRPC5 positive neurons in fixed murine trigeminal ganglia.

Validation Images

A. Representative image of ophthalmic region of trigeminal ganglion of C57Bl6/J mouse stained with ABIN2485379 diluted 1:500. B. Representative image of Z-Stack of ophthalmic region of trigeminal ganglion of TRPC5 ko stained with ABIN2485379 diluted 1:500.

Full Methods

Primary Antibody: ABIN2485379

Full Protocol:

• Surgical plane of anesthesia with Ketamine and Xylazine.
• Perfuse mouse transcardially with
• • 1ml/g BW ice cold PBS
  • 1ml/g BW 4% PFA.
• Postfix skull base in 4% PFA ON at 4°C.
• Harvest TGs in PBS.
• Wash TGs 4x with PBS for 30min at RT.
• Incubate TGs with rotator in blocking solution (PBS containing 1% BSA, 0.5% Triton X-100) ON at 4°C.
• Incubate TGs with rotator with primary FITC-conjugated mouse anti-TRPC5 antibody (antibodies-online, ABIN2485379, lot 1007) diluted 1:500 in blocking solution ON at 4°C.
• Wash TGs 4x with PBS for 30min at RT.
• Mount TGs with 1% low melting agarose.
• Whole Organ 2-Photon Imaging with a Zeiss LSM880 NLO Intravital Microscope; Settings: Excitation Wavelenth: 940 nm, Detection: 500-550 nm for wt and TRPC5 ko TGs.

Experimental Notes:

• In wt mice antibody concentrations of 1:100, 1:500, 1:1000 and 1:2000 were tested with best results using a concentration of 1:500.

• ABIN2485379 is suitable for whole organ IHC. TRPC5 positive Neurons show clear cytoplasmic and membranous staining. No neuronal staining visible. Residues of antibody visible in vasculature due to tissue manipulation during washing.

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: PBS pH 7.4, 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: 4 °C

Storage Comment: Conjugated antibodies should be stored at 4°C

Target Details for TRPC5

Target: TRPC5 (Transient Receptor Potential Cation Channel, Subfamily C, Member 5 (TRPC5))

Alternative Name: TRPC5

Background: Transient receptor potential cation channel, subfamily C, member 5, also known as TRPC5, is a subtype of the TRPC family of mammalian transient receptor potential ion channels. Homo-multimeric TRPC5 and hetero-multimeric TRPC5-TRPC1 channels are activated by extracellular reduced thioredoxin (1). This activation probably plays a role in rheumatoid arthritis. It has also been recently found to be involved in the action on anaesthetics such as chloroform, halothane and propofol (2).

Gene ID: 7224

NCBI Accession: NP_036603

UniProt: Q9UL62