Anti-PGK1 antibody [EPR19057]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PGK1 antibody [EPR19057] (AB199438)

Immunofluorescent analysis of 100% methanol-fixed Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling PGK1 with ab199438 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weak nuclear staining in Jurkat cells. The nuclear counterstain is DAPI (blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PGK1 antibody [EPR19057] (AB199438)

Immunofluorescent analysis of 100% methanol-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling PGK1 with ab199438 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weak nuclear staining in HeLa cells. The nuclear counterstain is DAPI (blue).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PGK1 antibody [EPR19057] (AB199438)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PGK1 with ab199438 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-PGK1 antibody [EPR19057] (AB199438)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PGK1 antibody [EPR19057] (ab199438) at 1/2000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-PGK1 antibody [EPR19057] (AB199438)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PGK1 antibody [EPR19057] (ab199438) at 1/2000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 20 µg

Lane 3:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-PGK1 antibody [EPR19057] (AB199438)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PGK1 antibody [EPR19057] (ab199438) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Rat brain lysate at 10 µg

Lane 5:

Rat heart lysate at 10 µg

Lane 6:

Rat kidney lysate at 10 µg

Lane 7:

C6 (Rat glial tumor cells) whole cell lysate

Lane 8:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate

Lane 9:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

Lane 10:

NIH/3T3 (Mouse embryo fibroblast cells) whole cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-PGK1 antibody [EPR19057] (AB199438)

Blocking and Diluting Buffer : 5% NFDM /TBST Lanes 3 and 4 Exposure : 60 seconds

Lanes 1 - 2:

Western blot - Anti-PGK1 antibody [EPR19057] (ab199438) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-PGK1 + PGK2 antibody [EPR14908] (<a href='/en-us/products/primary-antibodies/pgk1-pgk2-antibody-epr14908-ab186742'>ab186742</a>) at 1/5000 dilution

Lanes 1 and 3:

PGK1 Human Recombinant Protein at 20 µg

Lanes 2 and 4:

PGK2 Human Recombinant Protein at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 45 kDa

false

Exposure time: 5s

• WB

CiteAb

Western blot - Anti-PGK1 antibody [EPR19057] (AB199438)

PGK1 western blot using anti-PGK1 antibody [EPR19057] ab199438. Publication image and figure legend from Elkis, Y., Cohen, M., et al., 2017, Nat Commun, PubMed 29038547.

ab199438 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab199438 please see the product overview.

Development of the Fer/FerT kinase inhibitor-E260 by using a yeast-based HTS system. a Native Fer (pAES-Fer) and a mutated kinase inactive Fer (pAES-FerY715F) were expressed in yeast cells using the pAES expression vector. Protein lysates from transfected cells were subjected to WB analysis using anti-HA (for HA-tagged Fer), anti-Fer, anti-phosphotyrosine (p-tyr), and anti-actin antibodies. b Growth curve of yeast cells expressing either the empty pAES vector, native Fer, inactive Fer, and non-transformed yeast cells; n = 3,±SE. c Scheme of the HTS program. d The chemical structure the 0342 compound. e The chemical structure of the E260 compound. f Growth curve of yeast transformed with the different plasmids and treated with either DMSO, 2 µM E260, or 2 µM 0342. pAES-harboring cells treated with DMSO, mutant Fer treated with DMSO, native Fer-expressing cells treated with DMSO, native Fer-expressing cells treated with 0342, native Fer-expressing cells treated with E260. Untransformed yeast cells served as a negative control; n = 3, ±SE. g Histograms depicting the 600 nm OD readings of the yeast growth-curves (presented in f) after 40 h; n = 3, ±SE. h Protein lysates prepared from yeast expressing mutant Fer and treated with DMSO for 24 h (pAES-FerY715F), active Fer and treated with 2 µM E260 (pAES-Fer E260), active Fer and treated with DMSO (pAES-Fer DMSO), or harboring the pAES vector alone and treated with DMSO, were subjected to WB analysis using anti-Fer, anti-p-tyr, and anti-PGK antibodies. Arrow on the right depicts the MW of Fer. This image presents one experiment representative of three, which gave similar results

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