Antibodies, Proteins, Kits and Reagents for Life Science

Anti-RING2 / RING1B / RNF2 antibody [EPR12245] - BSA and Azide free (ab250376)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR12245] to RING2 / RING1B / RNF2 - BSA and Azide free
Suitable for: IHC-P, Flow Cyt (Intra), ICC, WB
Knockout validated
Reacts with: Mouse, Human

Product name

Anti-RING2 / RING1B / RNF2 antibody [EPR12245] - BSA and Azide free
See all RING2 / RING1B / RNF2 primary antibodies
Description

Rabbit monoclonal [EPR12245] to RING2 / RING1B / RNF2 - BSA and Azide free
Host species

Rabbit
Tested applications

Suitable for: IHC-P, Flow Cyt (Intra), ICC, WBmore details
Species reactivity

Reacts with: Mouse, Human
Predicted to work with: Rat
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- HeLa, 293, Jurkat and HepG2 whole cell lysate (ab7900); Human placenta tissue; HepG2 cells.
General notes
ab250376 is the carrier-free version of ab181140.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze.
Storage buffer

pH: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR12245
Isotype

IgG
Research areas

Western blot - Anti-RING2 / RING1B / RNF2 antibody [EPR12245] - BSA and Azide free (ab250376)

All lanes : Anti-RING2 / RING1B / RNF2 antibody [EPR12245] (ab181140) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RNF2 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 38 kDa
Observed band size: 42 kDa
why is the actual band size different from the predicted?

This data was developed using the same antibody clone in a different buffer formulation (ab181140).

Lanes 1- 2: Merged signal (red and green). Green - ab181140 observed at 42 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab181140 was shown to react with RING2 / RING1B / RNF2 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264845 (knockout cell lysate ab257640) lane below 42kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and RNF2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab181140 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RING2 / RING1B / RNF2 antibody [EPR12245] - BSA and Azide free (ab250376)

This data was developed using ab181140, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling RING2 / RING1B / RNF2 with ab181140 at 1/500 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin. Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Western blot - Anti-RING2 / RING1B / RNF2 antibody [EPR12245] - BSA and Azide free (ab250376)

This data was developed using ab181140, the same antibody clone in a different buffer formulation.

Lane 1: Wild-type HAP1 cell lysate (20 µg)

Lane 2: RING2 / RING1B / RNF2 knockout HAP1 cell lysate (20 µg)

Lane 3: HeLa cell lysate (20 µg)

Lane 4: Jurkat cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab181140 observed at 42 kDa. Red - loading control, ab7291, observed at 52 kDa.

ab181140 was shown to recognize RING2 / RING1B / RNF2 when RING2 / RING1B / RNF2 knockout samples were used, along with additional cross-reactive bands. Wild-type and RING2 / RING1B / RNF2 knockout samples were subjected to SDS-PAGE. ab181140 at a dilution of 1/1000 and ab7291 (loading control to alpha Tubulin) at a dilution of 1/10,000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.
Flow Cytometry - Anti-RING2 / RING1B / RNF2 antibody [EPR12245] - BSA and Azide free (ab250376)

This data was developed using ab181140, the same antibody clone in a different buffer formulation.Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling RING2 / RING1B / RNF2 with purified ab181140 at 1/70 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488)(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.
Western blot - Anti-RING2 / RING1B / RNF2 antibody [EPR12245] - BSA and Azide free (ab250376)

All lanes : Anti-RING2 / RING1B / RNF2 antibody [EPR12245] (ab181140) at 1/10000 dilution

Lane 1 : HeLa cell lysate
Lane 2 : 293 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HepG2 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 38 kDa

This data was developed using ab181140, the same antibody clone in a different buffer formulation.
Immunocytochemistry - Anti-RING2 / RING1B / RNF2 antibody [EPR12245] - BSA and Azide free (ab250376)

This data was developed using ab181140, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of HepG2 cells (paraformaldehyde-fixed, 4%) labeling RING2 / RING1B / RNF2 with ab181140 at 1/250 dilution followed by Goat anti rabbit IgG (Alexa Fluor® 555) secondary at 1/200 dilution and counter-stained with DAPI (blue).
Anti-RING2 / RING1B / RNF2 antibody [EPR12245] - BSA and Azide free (ab250376)

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