Antibodies, Proteins, Kits and Reagents for Life Science

318 288 ₸ Product size

100 µl 318 288 ₸

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Anti-LKB1 antibody [EPR19379] (ab199970)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19379] to LKB1
Suitable for: WB, IP
Knockout validated
Reacts with: Human

Product name

Anti-LKB1 antibody [EPR19379]
See all LKB1 primary antibodies
Description

Rabbit monoclonal [EPR19379] to LKB1
Host species

Rabbit
Tested Applications & Species

Application Species
IP
Human

WB
Human

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293T, Jurkat and K562 whole cell lysates; Human brain, human testis, fetal heart, fetal kidney and fetal testis lysates. IP: K562 and human testis lysates.
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.

Application	Species
IP	Human
WB	Human

Form

Liquid
Storage instructions

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

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19379
Isotype

IgG
Research areas

Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)

All lanes : Anti-LKB1 antibody [EPR19379] (ab199970) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : STK11 knockout HEK-293T cell lysate
Lane 3 : Hu testis cell lysate
Lane 4 : HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 45, 49 kDa
Observed band size: 55 kDa
why is the actual band size different from the predicted?

Lanes 1- 4: Merged signal (red and green). Green - ab199970 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab199970 was shown to react with LKB1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266647 (knockout cell lysate ab257712) was used. Wild-type HEK-293T and STK11 knockout HEK-293T 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. ab199970 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 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-LKB1 antibody [EPR19379] (ab199970)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: STK11 knockout HAP1 whole cell lysate (20 µg)
Lane 3: Hu Testis whole cell lysate (20 µg)

Lanes 1 - 3: Merged signal (red and green). Green - ab199970 observed at 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab199970 was shown to specifically react with STK11 in wild-type HAP1 cells. No band was observed when STK11 knockout samples were examined. Wild-type and STK11 knockout samples were subjected to SDS-PAGE. Ab199970 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)

All lanes : Anti-LKB1 antibody [EPR19379] (ab199970) at 1/1000 dilution

Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 45, 49 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

Negative control: HeLa (PMID 11297520; PMID24086281; PMID 19340305).
Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)

All lanes : Anti-LKB1 antibody [EPR19379] (ab199970) at 1/1000 dilution

Lane 1 : Human brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal testis lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 45, 49 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-LKB1 antibody [EPR19379] (ab199970)

LKB1 was immunoprecipitated from 1 mg of K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate with ab199970 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab199970 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: K562 whole cell lysate, 10 μg (Input).

Lane 2: ab199970 IP in K562 whole cell lysate.

Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab199970 in K562 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.

LKB1 isoforms: PMID 18854309.
Immunoprecipitation - Anti-LKB1 antibody [EPR19379] (ab199970)

LKB1 was immunoprecipitated from 1 mg of Human testis lysate with ab199970 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab199970 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: Human testis lysate, 10 μg (Input).

Lane 2: ab199970 IP in Human testis lysate .

Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab199970 in Human testis lysate .

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.

LKB1 isoforms: PMID 18854309.
Anti-LKB1 antibody [EPR19379] (ab199970)

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