MA5-14104

antibody from Invitrogen Antibodies

Targeting: ESR1 ER-alpha, Era, ESR, NR3A1

Antibody data

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Validation data

Product number: MA5-14104 - Provider product page
Provider: Invitrogen Antibodies
Product name: Estrogen Receptor alpha Antibody Cocktail
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-14104 is a mixture of two monoclonal antibodies that target Estrogen Receptor alpha. Staining of formalin-fixed, paraffin-embedded tissues requires boiling tissues in 10mM citrate buffer (pH 6.0) for 10-20 min, followed by cooling at room temperature for 20 min. A recommended positive control is MCF-7 cells.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 1D5, 6F11
Vial size: 500 µL
Concentration: Conc. Not Determined
Storage: 4° C

ESR1 protein structure - MA5-14104 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Estrogen Receptor alpha (ER alpha) 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 alpha was detected at ~66 kD (left panel) using an ER alpha monoclonal antibody (Product # MA5-14104) at a dilution of 1:500 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:40,000. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). Directly alongside the above blot, a second blot containing the same recombinant ER alpha and ER beta protein samples was probed with an ER-beta specific polyclonal antibody (Product # PA1-313, 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 alpha (ER) 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 alpha was detected at ~66 kD (top panel) using an ER alpha monoclonal antibody (Product # MA5-14104) at a dilution of 1:500 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) 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, a duplicate blot was 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 also performed using SuperSignal West Dura (Product # 34075).