702611

antibody from Invitrogen Antibodies

Targeting: LINC02210-CRHR1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

702611

Provider

Invitrogen Antibodies

Product name

CRHR1 Recombinant Rabbit Monoclonal Antibody (4H5L2)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Reactivity

Human, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

4H5L2

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on HEK-293 cells stably transfected with human CRHR1 (Lane 2) and mock (Lane 1). Receptors were eluted from the beads using SDS sample buffer for 20 min at 45 °C. Extracts were separated on 7.5% SDS-polyacrylamide gels and blotted onto PVDF membranes. The blot was probed with Anti-CRHR1 Recombinant Rabbit Monoclonal Antibody (Product # 702611, 1 µg/mL, 1:500 dilution). An ~80-100 kDa band corresponding to CRHR1 was observed only in the transfected cell line.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, HEK-293 cells were stably transfected with human CRHR1. After 24hR of transfection the cells were fixed and permeabilized for detection of CRHR1 using Anti-CRHR1 Recombinant Rabbit Monoclonal Antibody (Product # 702611, 1 µg/mL, 1:500 dilution) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate. Panel a) shows representative cells that were stained for detection of membrane localization of the CRHR1 on the transfected cells. The images were captured at 40X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunocytochemistry analysis, HEK 293 cells stably transfected with CRHR1 were grown on poly-L-lysine-coated coverslips overnight. Cells were then treated with 1 µM Corticotropin releasing hormone for 30 min, washed with PBS and fixed with 4% paraformaldehyde and 0.2% picric acid in phosphate buffer (pH 6.9) for 30 min at room temperature and washed several times with phosphate buffer. Specimens were permeabilized with ice-cold methanol and then incubated with Anti-CRHR1 Recombinant Rabbit Monoclonal Antibody (Product # 702611, 1 µg/mL). Cells were then incubated with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate for 2 h at room temperature, mounted and examined using a laser scanning confocal microscope. Note, robust receptor internalization after agonist exposure.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sections of Rat pituitary and hypothalamus was dewaxed, microwaved in citric acid and incubated with Anti-CRHR1 Recombinant Rabbit Monoclonal Antibody (Product # 702611, 1 µg/mL, 1:500 dilution). Sections were then sequentially treated with biotinylated Anti-rabbit IgG and AB solution. Sections were further developed in AEC and lightly counterstained with hematoxylin. Panel a & b) shows the specific localization of CRHR1 in neuronal cells in both Rat pituitary and hypothalamus respectively. Panel c) shows the no primary antibody control.