Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E277] to ELK1
Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
Knockout validated
Reacts with: Human

Product name

Anti-ELK1 antibody [E277]
See all ELK1 primary antibodies
Description

Rabbit monoclonal [E277] to ELK1
Host species

Rabbit
Specificity

This antibody does not cross react with other ETS family members

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
WB	Human

See all applications and species data

Immunogen

corresponding to Human ELK1 (C terminal).
Epitope

ab32106 reacts with an epitope located in the C terminal region of ELK1.
Positive control
- IHC-P: Human breast carcinoma ICC/IF: A549 cells WB: HeLa, MCF7 and HAP1 cell lysates. Flow Cyt: HeLa cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

E277
Isotype

IgG
Research areas

Western blot - Anti-ELK1 antibody [E277] (ab32106)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: ELK1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa positive whole cell lysate (21 µg)
Lane 4: MCF7 negative whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab32106 observed at 55 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab32106 was shown to specifically react with ELK1 in wild-type HAP1 cells as signal was lost in ELK1 knockout cells. Wild-type and ELK1 knockout samples were subjected to SDS-PAGE. Ab32106 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-ELK1 antibody [E277] (ab32106)

Immunocytochemistry/ Immunofluorescence analysis of A549 (human lung carcinoma epithelial cell) cells labeling ELK1 with purified ab32106 at 1/500 dilution (3.8 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ELK1 antibody [E277] (ab32106)

Immunohistochemical analysis of paraffin embedded human breast carcinoma using ab32106 at a dilution of 1/50

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cytometry - Anti-ELK1 antibody [E277] (ab32106)

Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ELK1 with unpurified ab32106 at 1/20 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
Western blot - Anti-ELK1 antibody [E277] (ab32106)

All lanes : Anti-ELK1 antibody [E277] (ab32106) at 1/500 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ELK1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 45 kDa
Observed band size: 55 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab32106 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab32106 was shown to react with ELK1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261764 (knockout cell lysate ab256904) was used. Wild-type HeLa and ELK1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32106 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-ELK1 antibody [E277] (ab32106)

Anti-ELK1 antibody [E277] (ab32106) at 1/500 dilution
Predicted band size: 45 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?
Anti-ELK1 antibody [E277] (ab32106)

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