Anti-G-CSF antibody [EPR3203(N)(B)]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-G-CSF antibody [EPR3203(N)(B)] (AB181053)

Intracellular Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) cells labeling G-CSF with purified ab181053 at 1/200 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-G-CSF antibody [EPR3203(N)(B)] (AB181053)

Immunocytochemistry/ Immunofluorescence analysis of BxPC-3 (Human pancreas adenocarcinoma epithelial cell) cells labeling G-CSF with Purified ab181053 at 1 : 500 dilution (4.0μg/ml). Cells were fixed in 100% Methanol. ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1 : 1000 dilution. DAPI nuclear was used as a counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-G-CSF antibody [EPR3203(N)(B)] (AB181053)

Immunofluorescent analysis of HT-1376 cells (paraformaldehyde-fixed, 4%) labeling G-CSF with unpurified ab181053 at 1/100 dilution followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary at 1/200 dilution and counter-stained with DAPI (blue).

• IP

Supplier Data

Immunoprecipitation - Anti-G-CSF antibody [EPR3203(N)(B)] (AB181053)

Western blot analysis of immunoprecipitation pellet from K562 cell lysate (lane 1) or a Negative control (lane 2) immunoprecipitated using unpurified ab181053 at 1/20 dilution.

Secondary : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.

All lanes:

Immunoprecipitation - Anti-G-CSF antibody [EPR3203(N)(B)] (ab181053)

Predicted band size: 22 kDa

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• IP

Lab

Immunoprecipitation - Anti-G-CSF antibody [EPR3203(N)(B)] (AB181053)

ab181053 (purified) at 1 : 100 dilution (2ug) immunoprecipitating G-CSF in K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate.
Lane 1 (input) : K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10ug
Lane 2 (+) : ab181053 & K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab181053 in K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST."

All lanes:

Immunoprecipitation - Anti-G-CSF antibody [EPR3203(N)(B)] (ab181053)

Predicted band size: 22 kDa

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• WB

Lab

Western blot - Anti-G-CSF antibody [EPR3203(N)(B)] (AB181053)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-G-CSF antibody [EPR3203(N)(B)] (ab181053) at 1/2000 dilution

Lane 1:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates at 20 µg

Lane 2:

HT-1376 (Human urinary bladder carcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

Mouse brain lysates at 20 µg

Lane 4:

Rat brain lysates 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: 22 kDa

Observed band size: 25 kDa

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• WB

Supplier Data

Western blot - Anti-G-CSF antibody [EPR3203(N)(B)] (AB181053)

All lanes:

Western blot - Anti-G-CSF antibody [EPR3203(N)(B)] (ab181053) at 1/2000 dilution

Lane 1:

K562 cell lysate at 20 µg

Lane 2:

KM3 cell lysate at 20 µg

Lane 3:

NCI-H460 cell lysate at 20 µg

Lane 4:

HT-1376 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L) HRP at 1/1000 dilution

Predicted band size: 22 kDa

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• WB

Lab

Western blot - Anti-G-CSF antibody [EPR3203(N)(B)] (AB181053)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 4 s

All lanes:

Western blot - Anti-G-CSF antibody [EPR3203(N)(B)] (ab181053) at 1/1000 dilution

Lane 1:

A549 (Human lung carcinoma epithelial cell) cell lysate at 10 µg

Lane 2:

K-562 (Human chronic myelogenous leukemia lymphoblast) cell lysate at 10 µg

Lane 3:

HepG2 (Human hepatocellular carcinoma epithelial cell) cell lysate at 10 µg

Lane 4:

HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate at 10 µg

Lane 5:

MCF7 (Human breast adenocarcinoma epithelial cell) cell lysate at 10 µg

Lane 6:

MOLT-4 (Human lymphoblastic leukemia T lymphoblast) cell lysate at 10 µg

Lane 7:

PC-3 (Human prostate adenocarcinoma epithelial cell) cell 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 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 25 kDa

false