- PA5-28752 - Invitrogen Antibodies GHSR antibody

Submitted references Defective dystrophic thymus determines degenerative changes in skeletal muscle.

Farini A, Sitzia C, Villa C, Cassani B, Tripodi L, Legato M, Belicchi M, Bella P, Lonati C, Gatti S, Cerletti M, Torrente Y
Nature communications 2021 Apr 8;12(1):2099

ASSOCIATION OF GHRELIN RECEPTOR AND INFLAMMATION IN PERI-ATRIAL ADIPOSE TISSUE FROM OBESE PATIENTS WITH POSTOPERATIVE ATRIAL FIBRILLATION.

Mocanu V, Timofte D, Oboroceanu T, Cretu-Silivestru IS, Pricope-Veselin A, Moraru M, Butcovan D
Acta endocrinologica (Bucharest, Romania : 2005) 2020 Jul-Sep;16(3):298-302

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of GHS-R1 using A) 30 µg NT2D1 whole cell lysate and B) 30 µg SK-N-SH whole cell lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a GHS-R1 polyclonal antibody (Product # PA5-28752) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of GHSR was performed by separating 50 µg of mouse tissue extract by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a GHSR Polyclonal Antibody (Product # PA5-28752) at a dilution of 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of GHSR was performed by separating 50 µg of various tissue extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a GHSR Polyclonal Antibody (Product # PA5-28752) at a dilution of 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of GHSR was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a GHSR Polyclonal Antibody (Product # PA5-28752) at a dilution of 1:500 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of GHSR was achieved by transfecting A549 with GHSR specific siRNAs (Silencer® select Product # s194432, s194431). Western blot analysis (Fig. a) was performed using whole cell extracts from the GHSR knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with GHSR Polyclonal Antibody (Product # PA5-28752, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to GHSR.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of A549 (Lane 1), U-87 MG (Lane 2), SH-SY5Y (Lane 3), Hep G2 (Lane 4), NTERA-2 (Lane 5), MDA-MB-231 (Lane 6) and tissue extract of Mouse Brain (Lane 7). The blot was probed with Anti-GHSR Polyclonal Antibody (Product # PA5-28752, 1:3000 dilution) and detected by chemiluminescence using Goat anti Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 50 kDa band corresponding to GHSR was observed across all cell lines and tissue extract tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of GHSR was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GHSR Polyclonal Antibody (Product # PA5-28752) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 Altered thymic architecture in mdx mice. Representative images of H&E staining of thymus of 3-month-old C57Bl and mdx mice revealed differences in medullary/cortex boundaries between animals (dashed white line) ( a ). WB analysis showed the downregulation of cytokeratin 14/16 in mdx thymus related to C57Bl ( b ). Thymic architecture of C57Bl and mdx mice characterized by immunofluorescence staining for cortical cytokeratin CK8 (green) and medullary cytokeratin CK5 (red) confirmed changes in dystrophic thymic environment. Graph displays fluorescence area % occupied by CK5 and CK8, as calculated by ImageJ software ( c ). Double immunofluorescence staining for CK5 (red) and CD3 (green) of C57Bl and mdx thymi portrayed a loosen embedding of CD3+ cells within dystrophic medulla. d Staining of ghrelin (GHR) and ghrelin receptor (GHS-R) (red) showed a comparable distribution of GHR between animals, but a prevalent expression of GHS-R in thymus of C57Bl mice. Of note, GHR was preferentially found in proximity of cortical CK8 (green). e Graph displays fluorescence area % occupied by GHS-R, as calculated by ImageJ software, in C57Bl and mdx mice ( f ). Expression of FoxP3+ cells (magenta) was evaluated by immunofluorescence staining within CK5+ thymic medulla (green). C-terminal containing dystrophin isoforms were detected by DYS-2 antibody (red) to identify a specific protein distribution within thymus. mTEC maturation level was evaluated by immunohistochemistry staining of C57

PA5-28752

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