Antibodies, Proteins, Kits and Reagents for Life Science

284 784 ₸ Product size

100 µl 284 784 ₸

Order now and get it on Tuesday March 02, 2021

Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19656] to HIF-2-alpha
Suitable for: IP, WB, ICC/IF
Reacts with: Human

Product name

Anti-HIF-2-alpha antibody [EPR19656]
See all HIF-2-alpha primary antibodies
Description

Rabbit monoclonal [EPR19656] to HIF-2-alpha
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Whole cell lysate of HeLa cells treated with 1mM deferoxamine (DFO) mesylate salt for 24h ; Whole cell lysate of HeLa cells treated with 100µM CoCl2 for 24h. ICC/IF: HeLa cells treated with DFO mesylate salt (1mM, 24h).
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19656
Isotype

IgG
Research areas

Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

All lanes : Anti-HIF-2-alpha antibody [EPR19656] (ab207607) at 1/1000 dilution

Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Whole cell lysate of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 1mM deferoxamine (DFO) mesylate salt for 24h

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 96 kDa
Observed band size: 110 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight observed is consistent with the literature, and the expression level can be induced under hypoxia or treated with hypoxia mimetic chemicals such as DFO (PMID: 25587023, 10934146, 23958427 and 21571835).
Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, untreated or treated with deferoxamine mesylate salt (1mM, 24h), labeling HIF-2-alpha with ab207607 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on HeLa cells treated with deferoxamine mesylate salt (1mM, 24h).

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

HIF-2-alpha was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell treated with 1mM DFO for 24h, cell lysate 10 µg with ab207607 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab207607 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate (10 µg) treated with 1mM DFO for 24h

Lane 2: ab207607 IP in DFO treated HeLa cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207607 in HeLa cell lysate

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

Exposure time: 75 seconds
Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

All lanes : Anti-HIF-2-alpha antibody [EPR19656] (ab207607) at 1/1000 dilution

Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Whole cell lysate of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 100µM CoCl2 for 24h

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 96 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight observed is consistent with the literature, and the expression level can be induced under hypoxia or treated with hypoxia mimetic chemicals such as CoCl₂ (PMID: 21571835, 10934146 and 23958427).
Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

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