PA5-19599

antibody from Invitrogen Antibodies

Targeting: HNRNPL HNRPL

Immunohistochemistry

Antibody data

Submit

Validation data

Product number: PA5-19599 - Provider product page
Provider: Invitrogen Antibodies
Product name: hnRNP L Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: This antibody is predicted to react with mouse based on sequence homology.
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 0.6 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

HNRNPL protein structure - PA5-19599 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HeLa Whole Cell Lysate using Product # PA5-19599, hnRNP L primary antibody at a dilution of 1 µg/mL. Blot treated with a secondary HRP-conjugated Goat polyclonal anti-Rabbit antibody was used at a dilution of 1:3000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of HNRNPL was achieved by transfecting HeLa with HNRNPL specific siRNAs (Silencer® select Product # s6740, s6741). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the HNRNPL knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with hnRNP L Polyclonal Antibody (Product # PA5-19599, 1 µg/ml) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 HNRPL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-hnRNP L Polyclonal Antibody (Product # PA5-19599) and a 58 kDa corresponding to HNRNPL was observed across cell lines and tissue extracts tested except Mouse Heart which is reported to be low. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), MCF-7 (Lane 3), Jurkat (Lane 4), NIH/3T3 (Lane 5) and tissue extracts of Mouse Heart (Lane 6), Mouse Thymus (Lane 7) and Mouse Lung (Lane 8) were electrophoresed using Novex® 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 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: Immunofluorescent staining of HeLa cells using Product # PA5-19599, anti-hnRNP L antibody. The cells were fixed with PFA (4%)for 10 minutes, permabilised TBS-T for 20 minutes and exposed to the primary antibody at a concentration of 1 µg/mL for 1 hour at room temp. A solution of BSA (1%), normal goat serum (10%) and glycine (0.3 M) was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody was a 448 fluorescence conjugated Goat anti-rabbit IgG (green) at a dilution of 1:1000. A WGA- 594 fluorescent conjugated stain was used to label plasma membranes (red) and the nuclei stain was DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of HeLa cells using Product # PA5-19599, anti-hnRNP L antibody. The cells were fixed with PFA (4%)for 10 minutes, permabilised TBS-T for 20 minutes and exposed to the primary antibody at a concentration of 1 µg/mL for 1 hour at room temp. A solution of BSA (1%), normal goat serum (10%) and glycine (0.3 M) was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody was a 448 fluorescence conjugated Goat anti-rabbit IgG (green) at a dilution of 1:1000. A WGA- 594 fluorescent conjugated stain was used to label plasma membranes (red) and the nuclei stain was DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of HeLa cells using Product # PA5-19599, anti-hnRNP L antibody. The cells were fixed with PFA (4%)for 10 minutes, permabilised TBS-T for 20 minutes and exposed to the primary antibody at a concentration of 1 µg/mL for 1 hour at room temp. A solution of BSA (1%), normal goat serum (10%) and glycine (0.3 M) was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody was a 448 fluorescence conjugated Goat anti-rabbit IgG (green) at a dilution of 1:1000. A WGA- 594 fluorescent conjugated stain was used to label plasma membranes (red) and the nuclei stain was DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of hnRNP L was performed using 70% confluent log phase 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 hnRNP L Rabbit Polyclonal Antibody (Product # PA5-19599) at 2 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) 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 merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.