Anti-G-CSF antibody [EPR3203(N)(B)] (ab181053)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3203(N)(B)] to G-CSF
- Suitable for: Flow Cyt, WB, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-G-CSF antibody [EPR3203(N)(B)]
See all G-CSF primary antibodies -
Description
Rabbit monoclonal [EPR3203(N)(B)] to G-CSF -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIP HumanWB MouseRatHuman -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A549, K-562, HepG2, HeLa, MCF7, MOLT-4, PC-3, KM3, NCI-H460 and HT-1376 cell lysates; Mouse and rat brain lysates. ICC/IF: BxPC-3 and HT-1376 cells. IP: G-CSF IP in K562 cell lysate. Flow Cyt: K562 cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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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
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3203(N)(B) -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-G-CSF antibody [EPR3203(N)(B)] (ab181053) at 1/1000 dilution
Lane 1 : A549 (Human lung carcinoma epithelial cell) cell lysate
Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) cell lysate
Lane 3 : HepG2 (Human hepatocellular carcinoma epithelial cell) cell lysate
Lane 4 : HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate
Lane 5 : MCF7 (Human breast adenocarcinoma epithelial cell) cell lysate
Lane 6 : MOLT-4 (Human lymphoblastic leukemia T lymphoblast) cell lysate
Lane 7 : PC-3 (Human prostate adenocarcinoma epithelial cell) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 22 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 4 s
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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.
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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.
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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 : Anti-G-CSF antibody [EPR3203(N)(B)] (ab181053) at 1/2000 dilution (unpurified)
Lane 1 : K562 cell lysate
Lane 2 : KM3 cell lysate
Lane 3 : NCI-H460 cell lysate
Lane 4 : HT-1376 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L) HRP at 1/1000 dilution
Predicted band size: 22 kDa
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All lanes : Anti-G-CSF antibody [EPR3203(N)(B)] (ab181053) at 1/2000 dilution (purified)
Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
Lane 2 : HT-1376 (Human urinary bladder carcinoma epithelial cell) whole cell lysates
Lane 3 : Mouse brain lysates
Lane 4 : Rat brain lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 22 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST
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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).
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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.
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