Antibody data
- Product number
- PA5-18822
- Provider
- Invitrogen Antibodies
- Product name
- Anti-14-3-3 theta Polyclonal Antibody
- Provider product page
- Invitrogen Antibodies - PA5-18822
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with canine, mouse and rat based on sequence homology.
- Reactivity
- Human
- Host
- Goat
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Supportive validation
- Submitted by
-
Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot staining of Human Hippocampus lysate using Product # PA5-18822 at a concentration of 0.03 µg/mL, the primary antibody incubation was 1 hour and the detection method was chemiluminescence.
Supportive validation
- Submitted by
-
Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of 14-3-3 theta was achieved by transfecting HeLa with 14-3-3 theta siRNAs (Silencer® select Product # s21599). Western blot analysis (Fig. a) was performed using whole cell extracts from the 14-3-3 theta knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with 14-3-3 theta recombinant polyclonal antibody (Product # PA5-18822, 1:1000 dilution) and Rabbit anti-Goat IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to 14-3-3 theta.
Supportive validation
- Submitted by
-
Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti- 14-3-3 theta Polyclonal Antibody (Product # PA5-18822) and a 28kDa band corresponding to 14-3-3 theta was observed across cell lines tested. Whole cell extracts (1%SDS) (30 µg lysate) of OVCAR-3 (Lane 1), SH-SY5Y (Lane 2) and HeLa (Lane 3) were electrophoresed using NuPAGE® 4-12 % Bis-Tris 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 (1:1000 dilution) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 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
- Immunofluorescence analysis of 14-3-3 theta was performed using HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with 14-3-3 theta Goat Polyclonal Antibody (Product # PA5-18822) at 5µg/ml in 0.1% BSA and incubated overnight at 4 degree and then labeled with Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11078) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the composite image showing cytoplasmic localization of 14-3-3 theta in HeLa cells . Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..
Supportive data in Antibodypedia
Data presented on provider website
