PA5-13541

antibody from Invitrogen Antibodies

Targeting: EEF1A1 EE1A1, EEF1A, EF1A, LENG7

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-13541

Provider

Invitrogen Antibodies

Product name

EEF1A1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with bovine, chicken, hamster, mouse, rat, rabbit and Xenopus and zebrafish based on sequence homology.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

400 µL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis using an EEF1A1 polyclonal antibody (Product # PA5-13541) in MCF-7 cell lysates (35 µg per lane).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of SK-OV-3 (Lane 1), HCT 116 (Lane 2), PC-3 (Lane 3), THP-1 (Lane 4) and K-562 (Lane 5). The blot was probed with Anti-EEF1A1 Polyclonal Antibody (Product # PA5-13541, 1:3000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 50 kDa band corresponding to EEF1A1 was observed across the cell lines tested.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of EEF1A1 was achieved by transfecting MCF7 with EEF1A1 specific siRNAs (Silencer® select Product # s4477, s4478 ). Western blot analysis (Fig. a) was performed using whole cell extracts from the EEF1A1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with EEF1A1 Polyclonal Antibody (Product # PA5-13541, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 EEF1A1.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of MCF-7 cells using an EEF1A1 polyclonal antibody (Product # PA5-13541) at a dilution of 1:10-50, followed by a fluor-conjugated goat anti-rabbit secondary antibody (green). Nuclei were stained with DAPI (blue).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of EEF1A1 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 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with EEF1A1 Rabbit Polyclonal Antibody(Product # PA5-13541) 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™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 nucleus and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis in formalin-fixed, paraffin-embedded human brain tissue using an EEF1A1 polyclonal antibody (Product # PA5-13541), followed by HRP-conjugated secondary antibody and DAB staining.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of NCI-H292 cells using an EEF1A1 polyclonal antibody (Product # PA5-13541) (bottom), compared to a negative control cell (top) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody