Myosin VIIA anticorps

N° du produit ABIN2435035

Détail du produit anti-Myosin VIIA anticorps

Antigène: Myosin VIIA (MYO7A)

Reactivité: Humain, Souris

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp Myosin VIIA est non-conjugé

Application: ELISA, Immunohistochemistry (IHC)

Purification: Affinity purification

Immunogène: Synthetic peptide of human MYO7A

Isotype: IgG

Information d'application

Indications d'application: IHC 1:25-1:100

Restrictions: For Research Use only

Independent Validation (IF)

Validation #103142 (Immunofluorescence)

by: Martinelli Lab, Neuroscience Department, UConn Health

No.: #103142

Date: 17.05.2018

Antigène: MYO7A

Numéro du lot: EC8591

Application validée: Immunofluorescence

Contrôle positif:

Organ of Corti tissue dissected from the cochlea of the inner ear, from a WT mouse, C57BL/6 strain, 10 weeks old.

Contrôle négative: Primary and secondary only antibody controls

Conclusion:

Passed. ABIN2435036 recognizes MYO7A with the expected expression pattern in murine organ of Corti tissue dissected from the cochlea of the inner ear.

Validation Images

In this photograph of the 3 rows of cochlear outer hair cells taken with 63x objective, MYO7A is labeled with ABIN2435036 in green, nuclei are labeled with DAPI (blue), and red marks the parvalbumin signal. Note that MYO7A-positive cells are adjacent to the parvalbumin-positive synapses, as expected since MYO7A is expressed in the outer hair cells. MYO7A signal can thus be used to assist in the identification and counting of outer hair cells.

Protocole

Anticorps primaire: ABIN2435036

Anticorps secondaire: anti-rabbit AF546 conjugated antibody (Life Technologies, A11035, lot 1812311)

Full Protocol:

• Dissect organ of Corti from the mouse cochlea in the inner ear as described in Maison, Liberman, and Liberman (2016).
• Cryoprotect and freeze/thaw to permeabilize the tissue:
• Transfer cochlear pieces to a 5ml disposable cup with approximately 1ml of 30% sucrose in 100mM phosphate buffer (PBS) at RT.
  • Incubate tissue on a shaker for 15min at RT.
  • Wash tissue with 30% sucrose in PBS at RT.
  • Incubate tissue on a shaker for 15min at RT.
  • Place cup on dry ice until contents freeze completely.
  • Allow cup to thaw at RT.
  • Pipet out the sucrose solution and wash tissue 3x for 15min with PBS containing 0.1% triton X-100 on a shaker at RT.
• Block tissue with blocking solution (5% goat serum containing 0.3% Triton-X) on a shaker for 30-60min at RT.
• Pipet out PBS + detergent and add blocking solution.
• Cut cap off a 1.5ml microcentrifuge tube and transfer tissue pieces in the blocking solution to the flipped upside-down cap.
• Incubate tissue in the flipped upside-down caps with 100µl primary
• rabbit anti-MYO7A antibody (antibodies-online, ABIN2435036, lot EC8591) diluted 1:100 in blocking solution ON at RT on an agitator.
  • chicken anti-parvalbumin (Synaptic Systems, 195006) diluted 1:400 in blocking solution ON at RT on an agitator.
• Fasten tubes onto the caps and and protect them from the light.
• Pipet out the primary antibody solution.
• Rinse tissue 3x for a total of 10min with PBS containing 0.1% Triton X-100.
• Incubate tissue in the flipped upside-down caps with 100µl secondary
• goat anti-rabbit AF546 conjugated antibody (Life Technologies, A11035, lot 1812311) diluted 1:300 in blocking solution for 1h at RT away from the light.
  • goat anti-chicken AF488 conjugated antibody (Life Technologies, A11039, lot 1812246) diluted 1:300 in blocking solution for 1h at RT away from the light.
• Rinse tissue 3x for a total of 10min with PBS.
• Transfer tissue pieces onto a slide with stereocilia facing up.
• Add mounting Fluoromount-G with DAPI mounting medium (ThermoFisher Scientific, 00-4959-52, lot B2215-N915) then coverslip.
• Image acquisition on a Zeiss Axiovert epifluorescence with with a 63x objective, using a Zeiss ApoTome for optical sectioning.

Notes:

• The observed signal on outer hair cells in the organ of Corti for ABIN2435036 appears as expected, compared to numerous other publications. For example, Figure 4 in He et al., and Figure 1 in Kaur et al., and Figure 4 in Li et al..

• ABIN2435036 worked well enough to detect the hair cells at 1:100 dilution. A more concentrated dilution would likely improve the signal to noise ratio.

• No signal was observed with either negative control.

Stockage

Format: Liquid

Concentration: 0.5 mg/mL

Buffer: PBS with 0.05 % sodium azide and 50 % glycerol, PH7.4

Agent conservateur: Sodium azide

Conseil sur la manipulation: Avoid freeze / thaw cycles.

Stock: -20 °C

Stockage commentaire: Store at -20°C. Avoid freeze / thaw cycles.

Détails sur Myosin VIIA

Antigène: Myosin VIIA (MYO7A)

Autre désignation: MYO7A (MYO7A Produits)

Synonymes: anticorps 53D10S, anticorps BG:DS00929.11, anticorps CG7595, anticorps D, anticorps D2, anticorps DM7a, anticorps DMyoVIIa, anticorps Dm 35B, anticorps DmVIIA, anticorps DmVIIa, anticorps Dmel\\CG7595, anticorps Dro35B, anticorps ESTS:53D10S, anticorps Mhc35BC, anticorps MyoVIIA, anticorps anon-35Bb, anticorps br27, anticorps ck/MyoVIIA, anticorps l(1)35Ca, anticorps l(2)07130, anticorps l(2)35Ca, anticorps l(2)br27, anticorps l35Ca, anticorps myoVIIA, anticorps stc, anticorps stch, anticorps myo7a, anticorps MYO7A, anticorps NV17859, anticorps Hdb, anticorps Myo7, anticorps USH1B, anticorps nmf371, anticorps polka, anticorps sh-1, anticorps sh1, anticorps DFNA11, anticorps DFNB2, anticorps MYOVIIA, anticorps MYU7A, anticorps NSRD2, anticorps crinkled, anticorps myosin VIIAa, anticorps myosin VIIa, anticorps myosin VIIA, anticorps myosin 7A, anticorps myosin-VIIa, anticorps ck, anticorps myo7aa, anticorps LOAG_02192, anticorps MYO7A, anticorps myo7a, anticorps LOC663995, anticorps LOC100116446, anticorps Myo7a

Sujet: This gene is a member of the myosin gene family. Myosins are mechanochemical proteins characterized by the presence of a motor domain, an actin-binding domain, a neck domain that interacts with other proteins, and a tail domain that serves as an anchor. This gene encodes an unconventional myosin with a very short tail. Defects in this gene are associated with the mouse shaker-1 phenotype and the human Usher syndrome 1B which are characterized by deafness, reduced vestibular function, and (in human) retinal degeneration. Alternative splicing results in multiple transcript variants.

NCBI Accession: NP_000251

Pathways: Sensory Perception of Sound