Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 02, 2021

Anti-TNF alpha antibody [EPR21753-109] (ab205587)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR21753-109] to TNF alpha
Suitable for: ICC/IF, WB, Flow Cyt, IP
Reacts with: Mouse, Rat

Product name

Anti-TNF alpha antibody [EPR21753-109]
See all TNF alpha primary antibodies
Description

Rabbit monoclonal [EPR21753-109] to TNF alpha
Host species

Rabbit
Specificity

The protein level of TNF alpha in normal samples is very weak. The TNF alpha expression must be stimulated.

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse
IP	Rat
WB	Mouse Rat

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Rat splenocytes treated with 20ng/ml PMA, 1µg/ml Ionomycin and 10µM BFA for 6h whole cell lysate; RAW 264.7 treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h, whole cell lysate. ICC/IF: RAW 264.7 cells. Flow: RAW 264.7 cells. IP: Rat splenocytes treated with 20ng/ml PMA, 1µg/ml Ionomycin and 10µM BFA for 6h, whole cell lysate
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR21753-109
Isotype

IgG
Research areas

Immunoprecipitation - Anti-TNF alpha antibody [EPR21753-109] (ab205587)

TNF alpha was immunoprecipitated from 0.35 mg rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate with ab205587 at 1/50 dilution. Western blot was performed on the immunoprecipitate using ab205587 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

Lane 1: Rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate 10μg (input).

Lane 2: ab205587 IP in rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab205587 in rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.

The MW observed is consistent with what has been described in the literature (PMID: 9933416).
Flow Cytometry - Anti-TNF alpha antibody [EPR21753-109] (ab205587)

Flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h (Red) / Untreated control (Green), compared to an unlabeled control (cells without incubation with primary antibody and secondary antibody, Blue) and an isotype control - Rabbit monoclonal IgG (ab172730, Black). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077)at 1/2000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR21753-109] (ab205587)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (+/- treatment with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h) cells labeling TNF alpha with ab205587 at 1/100 dilution, followed by a AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 cells treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h. The Nuclear counterstain is DAPI (Blue). Tubulin was stained using an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594, ab195889) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
Western blot - Anti-TNF alpha antibody [EPR21753-109] (ab205587)

All lanes : Anti-TNF alpha antibody [EPR21753-109] (ab205587) at 1/1000 dilution

Lane 1 : Untreated rat splenocytes whole cell lysate
Lane 2 : Rat splenocytes treated with 20ng/ml PMA, 1µg/ml Ionomycin and 10µM BFA for 6h whole cell lysate
Lane 3 : Untreated RAW 264.7(mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 4 : RAW 264.7 treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 26 kDa

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

Exposure time: 20 seconds

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID: 9933416)
Anti-TNF alpha antibody [EPR21753-109] (ab205587)

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