Anti-GATA2 antibody [EPR2822(2)]

• IP

Unknown

Immunoprecipitation - Anti-GATA2 antibody [EPR2822(2)] (AB109241)

GATA2 was immunoprecipitated from 0.35 mg K-562 (Human chronic myelogenous leukemia lymphoblast) cell lysate 10 µg with ab109241 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab109241 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) cell lysate 10 µg

Lane 2 : ab109241 IP in K-562 cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab109241 in K562 cell lysate

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

Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-GATA2 antibody [EPR2822(2)] (ab109241)

Predicted band size: 50 kDa

false

• ChIP

Lab

ChIP - Anti-GATA2 antibody [EPR2822(2)] (AB109241)

Chromatin was prepared from K-562 (Human chronic myelogenous leukemia lymphoblast) cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then 1% formaldehyde for 10min.

The ChIP was performed with 25 µg of chromatin, 5 µg of ab109241 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR ( Sybr green approach).

Primers and probes are from paper PMID : 22036566.

• IP

Unknown

Immunoprecipitation - Anti-GATA2 antibody [EPR2822(2)] (AB109241)

Purified ab109241 at 1/30 dilution (2 μg) immunoprecipitating GATA2 in K-562 whole cell lysate.
Lane 1 (input) : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10 μg
Lane 2 (+) : ab109241 + K-562 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab109241 in K-562 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 51 kDa

All lanes:

Immunoprecipitation - Anti-GATA2 antibody [EPR2822(2)] (ab109241)

Predicted band size: 33 kDa,50 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-GATA2 antibody [EPR2822(2)] (AB109241)

Blocking and diluting buffer : 5% NFDM/TBST.

The different result in K-562 is due to the lysates preparation method.

For Lysate preparation protocol, please refer to the protocol section of the website and/or here.

The expression profile observed in U-937 is consistent with the literature (PMID : 19212333).
Negative control : U-937 (PMID : 19212333)

All lanes:

Western blot - Anti-GATA2 antibody [EPR2822(2)] (ab109241) at 1/1000 dilution

Lane 1:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate prepared using 1% SDS hot lysis method at 20 µg

Lane 2:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate prepared using RIPA lysis method at 20 µg

Lane 3:

U-937 (Human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 50 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-GATA2 antibody [EPR2822(2)] (AB109241)

Lanes 1 - 2 : Merged signal (red and green). Green - ab109241 observed at 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab109241 was shown to recognize GATA2 in wild-type HEK-293 cells as signal was lost at the expected MW in GATA2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GATA2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab109241 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GATA2 antibody [EPR2822(2)] (ab109241) at 1 µg/mL

Lane 1:

Wild-type HEK-293 whole cell lysate at 20 µg

Lane 2:

GATA2 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

Western blot - Human GATA2 knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-gata2-knockout-hek-293-cell-line-ab261720'>ab261720</a>)

Predicted band size: 50 kDa

false

• WB

Unknown

Western blot - Anti-GATA2 antibody [EPR2822(2)] (AB109241)

All lanes:

Western blot - Anti-GATA2 antibody [EPR2822(2)] (ab109241) at 1/1000 dilution

Lane 1:

bEnd.3 (Mouse brain endothelioma) whole cell lysate at 15 µg

Lane 2:

Mouse placenta lysate at 15 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 15 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: 50 kDa

Observed band size: 51 kDa

false

• WB

CiteAb

Western blot - Anti-GATA2 antibody [EPR2822(2)] (AB109241)

GATA2 western blot using anti-GATA2 antibody [EPR2822(2)] ab109241. Publication image and figure legend from Lu, W., Jia, D., et al., 2017, Sci Rep, PubMed 28827748.

ab109241 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 ab109241 please see the product overview.

CSEI regulated the expressions of hematopoietic function related proteins in CHRF and K562 cells. After a 24-h CSEI (0, 0.05, 0.1 mg/ml) treatment, the protein expression levels of P-RSK1p90, ELK1 and c-Myc in CHRF cells (a) and K562 cells (b), full-length blots are presented in Supplementary Fig. S5 a, and the expression levels of GATA-1 and GATA-2 in the terminal mature CHRF cells (c) and differentiated K562 cells (d) were detected by western blot, respectively, full-length blots are presented in Supplementary Fig. S5 (b). The quantitative data of these protein levels were normalized by their GAPDH expressions and expressed as a percentage of the corresponding relative intensity of control. The data are shown as the means ± S.E.M. (n = 6). *p < 0.05, **p < 0.01 and ***p < 0.001 versus 0 mg/ml CSEI-treated cells.

false