PA1-310B

antibody from Invitrogen Antibodies

Targeting: ESR2 ER-beta, Erb, NR3A2

Provider product page for PA1-310B

Western blot

Immunocytochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [80]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [1]
Other assay [6]

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Product number: PA1-310B - Provider product page
Provider: Invitrogen Antibodies
Product name: Estrogen Receptor beta Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: PA1-310B has been successfully used in Western blot, immunohistochemistry, immunofluorescence, and immunocytochemistry procedures. By Western blot, this antibody detects an ~55 kDa protein representing ER beta from both rat brain homogenate as well as various breast cancer cell lines. Immunocytochemical staining of ER beta in cells overexpressing the human protein results in nuclear staining.
Concentration: 1 mg/mL

ESR2 protein structure - PA1-310B shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Estrogen Receptor beta (ER beta) was performed by loading the indicated amounts of recombinant full-length human ER alpha (Product # A15674) and ER beta (Product # A15664) proteins, and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. ER beta was detected at ~54 kD (left panel) using an ER beta polyclonal antibody (Product # PA1-310B) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). The blot was then stripped with Restore Plus Western blot stripping buffer (Product # 46430) for at least 15 minutes at room temperature, re-blocked with 5% milk in TBST for at least 1 hour at room temperature, and re-probed with an ER alpha polyclonal antibody (Product # PA1-308, right panel) at a dilution of 1:2000 overnight at 4C, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000. Chemiluminescent detection was again performed using SuperSignal West Pico (Product # 34080).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Estrogen Receptor beta (ER beta) was performed by loading 20 µg of whole cell lysates (WCL) or nuclear extracts (NE) from the indicated breast cancer cell lines, and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. ER beta was detected at ~55 kD (top panel) using an ER beta polyclonal antibody (Product # PA1-310B) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075). To check for equivalent protein loading in each lane, the blot was stripped with Restore Plus Western blot stripping buffer (Product # 46430) for at least 15 minutes at room temperature, re-blocked with 5% milk in TBST for at least 1 hour at room temperature, and re-probed with an HDAC1 polyclonal antibody (Product # PA1-860, bottom panel) at a dilution of 1:1000 overnight at 4C, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000. Chemiluminescent detection was again performed using SuperSignal West Dura (Product # 34075).