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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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- Product number
- 702611 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CRHR1 Recombinant Rabbit Monoclonal Antibody (4H5L2)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with Monkey, Dog, Pig, Rabbit.
- Antibody clone number
- 4H5L2
- Concentration
- 0.5 mg/mL
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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.
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.
- 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.
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.
