MA1-722

antibody from Invitrogen Antibodies

Targeting: RHO CSNBAD1, OPN2, RP4

Antibody data

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Validation data

Product number: MA1-722 - Provider product page
Provider: Invitrogen Antibodies
Product name: Rhodopsin Monoclonal Antibody (1D4)
Antibody type: Monoclonal
Antigen: Other
Description: MA1-722 detects rhodopsin from human, mouse and bovine retinal samples. Clone 1D4 has also been used successfully in Zebrafish. It does not recognize rhodopsin in zebrafish rods, but instead labels long double cone outer segments that express red opsin (Yin, J. et al, 2012).
Antibody clone number: 1D4
Concentration: 1 mg/mL

RHO protein structure - MA1-722 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Rhodopsin (green) showing staining in human retinal tissue (right) compared to a negative control without primary antibody (left). Formalin-fixed tissue was permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Tissue was probed with a Rhodopsin monoclonal antibody (Product # MA1-722) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4ºC in a humidified chamber. Tissue was washed with PBST and incubated with a DyLight-488 conjugated secondary antibody in PBS at room temperature in the dark. Nuclei were stained with DAPI (blue) and images were taken at a magnification of 20x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Rhodopsin showing staining in the nucleus of paraffin-embedded human retinal 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 Rhodopsin monoclonal antibody (Product # MA1-722) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Rhodopsin (green) showing staining in mouse retinal tissue (right) compared to a negative control without primary antibody (left). Formalin-fixed tissue was permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Tissue was probed with a Rhodopsin monoclonal antibody (Product # MA1-722) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4ºC in a humidified chamber. Tissue was washed with PBST and incubated with a DyLight-488 conjugated secondary antibody in PBS at room temperature in the dark. Nuclei were stained with DAPI (blue) and images were taken at a magnification of 20x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Rhodopsin showing staining in the nucleus of paraffin-embedded mouse retinal 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 Rhodopsin monoclonal antibody (Product # MA1-722) diluted in 3% BSA-PBS at a dilution of 1:1000 overnight at 4°C 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.

Supportive validation

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 4-PBA suppressed light-induced photoreceptor histological changes and degeneration. ( A ) Representative images of immunohistochemistry for rhodopsin (green) counterstained with 4',6-diamidino-2-phenylindole (DAPI) (blue) 4 days after light exposure with or without 4-PBA treatment. Reduction in ONL thickness ( B ) and OS length ( C ) were suppressed by 4-PBA. LE, light exposure; Veh, vehicle; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer; OS, outer segment. Data are presented as mean +- standard deviation. n = 8 in each group. Scale bar, 25 mum. * p < 0.05, ** p < 0.01, *** p < 0.001. ANOVA with Tukey post-hoc tests.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Protective effect of PBA in the photoreceptors of P23H RP models detected before photoreceptor loss ( A , A' ) Immunohistochemistry for Tom20 (green) and rhodopsin (pink) in the retina of P23H RP models at the age of 4 weeks with or without PBA treatment. Nuclei were counterstained with DAPI (blue). ( A ') Magnified images of IS and OS. IS area ( B ), and mRNA levels of rhodopsin ( C ) and crx ( D ) in the retina were already different with or without PBA treatment. Data are shown as mean +- standard deviation. Respective n for P23H RP models treated with vehicle and PBA, ( B ) 4 and 5; ( C ) 5 and 4; ( D ) 10 and 10. ONL, outer nuclear layer; IS, inner segments; OS, outer segments. * p < 0.05. Scale bar, ( A ) 20 mum; ( B ) 10 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Detection of rhodopsin levels in the retinas of all three mouse groups. (A) Western blot analysis was performed at P15 and the retinal protein extracts were run in SDS polyacrylamide gel and transferred to membrane. Images of membrane probed with anti-rhodopsin (upper) and anti-actin (lower) antibodies are shown. (B) The level of rhodopsin was calculated in all three groups. (C) Rhodopsin staining of retinal sections from the three groups at P15. Red, rhodopsin; blue, DAPI. Error bars: SEM. Statistical significance denoted by *P < 0.05, **P < 0.01, ****P < 0.001.