PA1-811

antibody from Invitrogen Antibodies

Targeting: RARB HAP, NR1B2, RRB2

Antibody data

Submit

Validation data

Product number: PA1-811 - Provider product page
Provider: Invitrogen Antibodies
Product name: RARB Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: PA1-811 detects retinoic acid receptor (RAR) beta from human and mouse tissues. This antibody shows slight cross-reactivity to RAR alpha but does not detect RAR gamma. PA1-811 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~52 kDa protein representing RAR beta from SH-SY5Y (human neuroblastoma) cell extract. The PA1-811 immunogen is a synthetic peptide corresponding to residues P(429) S V S P S S V E N S G V S Q S P L L Q(448) of mouse RAR beta. This immunizing peptide (Cat. # PEP-005) is available for use in neutralization and control experiments.
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: Conc. Not Determined
Storage: -20° C, Avoid Freeze/Thaw Cycles

RARB protein structure - PA1-811 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Retinoic Acid Receptor beta was performed by loading 25 µg of SH-SY5Y (lane 1), HepG2 (lane 2) and mouse heart (lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a Retinoic Acid Receptor beta polyclonal antibody (Product # PA1-811) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~58 kDa in SH-SY5Y and HepG2 cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-RARB Polyclonal Antibody (Product # PA1-811) and 58, 50 kDa bands corresponding to RARB were observed in SH-SY5Y and HEK-293 and not seen in PANC-1, OVCAR3 and BeWo which are reported to be negative. Whole cell extracts(30 µg lysate) of SH-SY5Y (Lane 1), HEK-293 (Lane 2), PANC-1 (Lane 3), OVCAR-3 (Lane 4) and BeWo (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/ml) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), Mouse Prostate (Lane 2) and MCF-7 (Lane3). The blots were probed with Anti-Retinoic Acid Receptor beta Rabbit Polyclonal Antibody (Product # PA1-811, 1:1000 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution).An isoform of 53kDa was observed in SH-SY5Y cell line, a 38kDa band was observed in Mouse Prostate tissue and around 50kDa band was observed in MCF-7 cell line tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0341BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Retinoic Acid Receptor beta was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Retinoic Acid Receptor beta Rabbit Polyclonal Antibody (Product # PA1-811) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of RARB was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled with RARB Polyclonal Antibody (Product # PA1-811) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034, 1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and cytoplasmic localization. Panel e represents BeWo having no expression of RARB. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.