Protocadherin-15 Antibody [Unconjugated]

Western blot shows lysates of HEK293T human embryonic kidney cell line and HEK293T cell line spiked with Recombinant Human Protocadherin-15 (5 ng/lane). PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human Protocadherin-15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6729) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Protocadherin-15 at approximately 190 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Protocadherin-15 was detected in immersion fixed YT human leukemia natural killer-like cell line using Sheep Anti-Human Protocadherin-15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6729) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Protocadherin-15 was detected in immersion fixed paraffin-embedded sections of human kidney using Sheep Anti-Human Protocadherin-15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6729) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in convoluted tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Hu
• Applications: WB, IHC, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Protocadherin-15 Antibody [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human Protocadherin-15
  Gln27-Ala1376
  Accession # Q96QU1
• Specificity: Detects human Protocadherin-15 in direct ELISAs.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: PCDH15
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Protocadherin-15 Antibody [Unconjugated]

• autosomal recessive 23
• cadherin-related family member 15
• CDHR15
• DFNB23
• DKFZp667A1711
• PCDH15
• Protocadherin15
• Protocadherin-15
• protocadherin-related 15
• USH1F

Background

Protocadherin 15 (PCDH-15) is a 250-270 kDa "other" member of the nonclustered group of the protocadherin family of molecules. In the adult, it is expressed on the base of hair cell stereoclila in the Organ of Corti, on cerebellar granule cells, and on rod and cone photoreceptors. In the ear, it is expressed as a homodimer, and binds (in trans) to cadherin 23 homodimers on adjacent stereocilia. Mature human PCDH-15 is a type I transmembrane protein that is 1929 amino acids (aa) in length. It contains a 1350aa extracellular domain (ECD) (aa 27-1376) plus a 558 aa cytoplasmic region. There are 11 cadherin domains in the ECD (aa 40-1259). Multiple splice forms are reported. One is a soluble 110 kDa form that shows a five aa substitution for aa 957-1955. Another contains a seven aa substitution for aa 1119-1125. Others show substitutions or deletions in either the extracellular or cytoplasmic domains. Over aa 27-1376 (the ECD), human PCDH-15 shares 95% aa identity with mouse PCDH-15.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Protocadherin-15 Antibody (AF6729)(5)

Publications using AF6729

• Ivanchenko, MV;Hathaway, DM;Mulhall, EM;Booth, KT;Wang, M;Peters, CW;Klein, AJ;Chen, X;Li, Y;György, B;Corey, DP; PCDH15 Dual-AAV Gene Therapy for Deafness and Blindness in Usher Syndrome Type 1F bioRxiv : the preprint server for biology 2023-11-13 [PMID: 38014037] (Immunohistochemistry, Primate)
• Verschueren A, Boucherit L, Ferrari U et al. Planar polarity in primate cone photoreceptors: a potential role in Stiles Crawford effect phototropism Communications Biology 1/24/2022 [PMID: 35075261]
• Verschueren A, Boucherit L, Ferrari U et al. Planar polarity in primate cone photoreceptors: a potential role in Stiles Crawford effect phototropism Communications Biology 2022-01-24 [PMID: 35075261]
• Wei Huang, Aparna Bhaduri, Dmitry Velmeshev, Shaohui Wang, Li Wang, Catherine A. Rottkamp et al. Origins and Proliferative States of Human Oligodendrocyte Precursor Cells Cell 8/1/2020 [PMID: 32679030]
• C Schietroma, K Parain, A Estivalet, A Aghaie, J Boutet de, S Picaud, JA Sahel, M Perron, A El-Amraoui, C Petit Usher syndrome type 1-associated cadherins shape the photoreceptor outer segment J. Cell Biol., 2017-05-11;0(0):. 2017-05-11 [PMID: 28495838] (IHC-Fr, Xenopus)

Reviews for Protocadherin-15 Antibody (AF6729) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Human, Mouse.

reviewed by: Verified Customer WB Human 10/02/2018

Summary

• Application: Western Blot
• Sample Tested: HEK 293 transfected,HEK293 transfected cells
• Species: Human
• Lot: 1806A

Comments

• Comments: PVDF membrane was blocked with 3% milk for 1 hour at room temperature. Primary antibody sheep anti-Pcdh15 was used at a concentration 2ug/mL in 3% milk and the membrane incubated overnight at 4C. Secondary antibody anti-sheep-HRP was used at a dilution 1:2500 in 3% milk and incubated for 1 hour at room temperature.

reviewed by: Verified Customer IHC-Fr Mouse 09/25/2018

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: Organ of Corti (inner ear)
• Species: Mouse
• Lot: 1806A

Comments

• Comments: P1 mouse organ of Corti was paraformaldehyde fixed (4%) for 30 min, blocked 1 hour in PBS Triton X-100 0.1% containing 10% fetal calf serum and then incubated overnight with sheep anti-Pcdh15 (1:200, green) in blocking solution. Secondary antibody was Alexa488 anti-sheep (1:200). Tissue was counterstained with phalloidin (red).

Bioinformatics

• Gene Symbol: PCDH15
• Uniprot:
  • Q96QU1(Human)