Invitrogen

JPH1 Polyclonal Antibody

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JPH1 Antibody — Published figure using JPH1 polyclonal antibody (Product # 40-5100)

JPH1 Antibody in Immunofluorescence (IF) — Immunofluorescent staining of mouse skeletal muscle using Junctophilin-1 (C-term) Polyclonal Antibody, Rabbit (Product # 40-5100). Image courtesy of James I. Nagy, PhD, University of Manitoba, Canada

JPH1 Antibody in Western Blot (WB) — Western blot analysis was performed on tissue extracts (30 µg lysate) of Mouse Muscle (Lane 1), and Mouse Heart (lane 2). The blots were probed with Anti-Junctophilin-1 Rabbit Polyclonal Antibody (Product # 40-5100, 2 µg/mL) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). A 70 kDa band corresponding to Junctophilin-1 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001).The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

JPH1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Junctophilin-1 showing staining in the cytoplasm of paraffin-embedded mouse skeletal muscle tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Anti- Junctophilin-1 Polyclonal Antibody (Product # 40-5100) diluted in 3% BSA-PBS at a dilution of 1:50 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Product Details

Tested Applications

Dilution

Immunofluorescence (IF)

Assay Dependent

Immunohistochemistry (Frozen) (IHC (F))

Assay Dependent

Immunohistochemistry (Paraffin) (IHC (P))

1:50

Western Blot (WB)

2-3 µg/mL

Published Applications

Western Blot (WB)

See 5 publications below

Immunohistochemistry (IHC)

See 2 publications below

Immunocytochemistry (ICC)

See 1 publication below

Immunohistochemistry (Paraffin) (IHC (P))

See 1 publication below

Immunofluorescence (IF)

See 1 publication below

Miscellaneous PubMed (MISC)

See 1 publication below

Product Specifications

Species Reactivity

Mouse, Rat

Published species

Human, Mouse, Rabbit, Rat

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide derived from the C-terminal region of the mouse junctophilin-1 (JP-1, junctophilin type 1, JPH1) protein, which differs from rat by one non-conservative amino acid replacement

Conjugate

Unconjugated

Form

Liquid

Concentration

0.25 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7.4

Contains

0.1% sodium azide

Storage conditions

-20°C

RRID

AB_2533470

Target Information

Junctional complexes between the plasma membrane (PM) and endoplasmic/sarcoplasmic reticulum (ER/SR) are a common feature of all excitable cell types and mediate cross talk between cell surface and intracellular ion channels. Junctophilins (JPs) are important components of the junctional complexes. JPs are composed of a carboxy-terminal hydrophobic segment spanning the ER/SR membrane and a remaining cytoplasmic domain that shows specific affinity for the PM. Four JPs have been identified as tissue-specific subtypes derived from different genes: JPH1 is expressed in skeletal muscle, JPH2 is detected throughout all muscle cell types, and JPH3 and JPH4 are predominantly expressed in the brain and contribute to the subsurface cistern formation in neurons. JPH1 is essential for stabilizing the T-tubule and SR membranes to form junctions and provide an environment for the assembly of receptors such as the ryanodine receptor type 1 (RyR1).

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.