Human Protocadherin-15 Antibody

R&D Systems | Catalog # AF6729

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Frog - Xenopus (African Clawed Frog), Primate

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all Protocadherin-15 Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human Protocadherin-15
Gln27-Ala1376
Accession # Q96QU1

Specificity

Detects human Protocadherin-15 in direct ELISAs.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human Protocadherin-15 Antibody

Detection of Human Protocadherin-15 antibody by Western Blot.

Detection of Human Protocadherin-15 by Western Blot.

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 antibody in YT Human Cell Line by Immunocytochemistry (ICC).

Protocadherin-15 in YT Human Cell Line.

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 antibody in Human Kidney by Immunohistochemistry (IHC-P).

Protocadherin‑15 in Human Kidney.

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.

Applications for Human Protocadherin-15 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed YT human leukemia natural killer-like cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human kidney

Western Blot

2 µg/mL
Sample: HEK293T human embryonic kidney cell line spiked with Recombinant Human Protocadherin-15

Reviewed Applications

Read 2 reviews rated 5 using AF6729 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Protocadherin-15

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.

Alternate Names

CDHR15, DFNB23, PCDH15, Protocadherin15, USH1F

Entrez Gene IDs

65217 (Human); 11994 (Mouse); 690865 (Rat)

Gene Symbol

PCDH15

UniProt

Q96QU1 (Human)

Additional Protocadherin-15 Products

Product Documents for Human Protocadherin-15 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Protocadherin-15 Antibody

For research use only

Related Research Areas

Cadherin Superfamily

Citations for Human Protocadherin-15 Antibody

Customer Reviews for Human Protocadherin-15 Antibody (2)

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2 Customer Ratings
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  • Human Protocadherin-15 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HEK293 transfected cells and HEK 293 transfected
    Species: Human
    Verified Customer | Posted 10/02/2018
    HEK 293 cells untransfected ;UT; or expressing Pcdh15-CD1 or Pcdh15-CD2 isoforms. A Pcdh15 construct the cytoplasmic tail was also expressed in these cells Ex;.
    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.
    Human Protocadherin-15 Antibody AF6729
  • Human Protocadherin-15 Antibody
    Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Organ of Corti (inner ear)
    Species: Mouse
    Verified Customer | Posted 09/25/2018
    P1 organ of Corti stained with sheep anti-Pcdh15 (green) and counterstained with phalloidin (red). Outer hair cells show a strong immunereactivity for Pcdh15 at the hair cell bundle level.
    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).
    Human Protocadherin-15 Antibody AF6729

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Protocols

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