Anti-G-CSF antibody [EPR3203(N)(B)] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-G-CSF antibody [EPR3203(N)(B)] - BSA and Azide free (AB236157)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181053).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-G-CSF antibody [EPR3203(N)(B)] - BSA and Azide free (AB236157)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181053).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-G-CSF antibody [EPR3203(N)(B)] - BSA and Azide free (AB236157)

Intracellular Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) cells labeling G-CSF with purified ab181053 at 1/200 dilution (10μg/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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181053).

• IP

Supplier Data

Immunoprecipitation - Anti-G-CSF antibody [EPR3203(N)(B)] - BSA and Azide free (AB236157)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181053).

All lanes:

Immunoprecipitation - Anti-G-CSF antibody [EPR3203(N)(B)] (<a href='/en-us/products/primary-antibodies/g-csf-antibody-epr3203nb-ab181053'>ab181053</a>)

Predicted band size: 22 kDa

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• IP

Lab

Immunoprecipitation - Anti-G-CSF antibody [EPR3203(N)(B)] - BSA and Azide free (AB236157)

ab181053 (purified) at 1 : 100 dilution (2μg) 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 10μg
Lane 2 (+) : ab181053 and 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181053).

All lanes:

Immunoprecipitation - Anti-G-CSF antibody [EPR3203(N)(B)] (<a href='/en-us/products/primary-antibodies/g-csf-antibody-epr3203nb-ab181053'>ab181053</a>)

Predicted band size: 22 kDa

false

• WB

Lab

Western blot - Anti-G-CSF antibody [EPR3203(N)(B)] - BSA and Azide free (AB236157)

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

Exposure time : 4 s

All lanes:

Western blot - Anti-G-CSF antibody [EPR3203(N)(B)] (<a href='/en-us/products/primary-antibodies/g-csf-antibody-epr3203nb-ab181053'>ab181053</a>) 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