Antibodies, Proteins, Kits and Reagents for Life Science

278 083 ₸ Product size

100 µl 278 083 ₸

Order now and get it on Tuesday March 02, 2021

Anti-RUNX3 antibody [EPR20680] (ab224642)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20680] to RUNX3
Suitable for: WB, IHC-P, IP
Knockout validated
Reacts with: Human

Product name

Anti-RUNX3 antibody [EPR20680]
See all RUNX3 primary antibodies
Description

Rabbit monoclonal [EPR20680] to RUNX3
Host species

Rabbit

Tested Applications & Species

Application	Species
IHC-P	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: Raji and Ramos whole cell lysates; Human thymus and tonsil lysates. IHC-P: Human stomach, gastric cancer and diffuse large B cell lymphoma tissues. IP: Raji 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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20680
Isotype

IgG
Research areas

Western blot - Anti-RUNX3 antibody [EPR20680] (ab224642)

All lanes : Anti-RUNX3 antibody [EPR20680] (ab224642) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : RUNX3 knockout HAP1 whole cell lysate
Lane 3 : Ramos whole cell lysate
Lane 4 : HeLa whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 44, 46 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab224642 observed at 44-46 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab224642 was shown to recognize RUNX3 in wild-type HAP1 cells as signal was lost at the expected MW in RUNX3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and RUNX3 knockout samples were subjected to SDS-PAGE. Ab224642 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-RUNX3 antibody [EPR20680] (ab224642)

All lanes : Anti-RUNX3 antibody [EPR20680] (ab224642) at 1/1000 dilution

Lane 1 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
Lane 3 : MCF7 (human breast adenocarcinoma cell line) 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

Developed using the ECL technique.

Predicted band size: 44, 46 kDa
Observed band size: 44,46 kDa
why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking/dilution buffer: 5% NFDM/TBST.

Negative control: : MCF-7 (PMID:21706051).
This target detects both predicted isoforms 44KDa and 46KDa, consistent with what has been described in the literature (PMID:23700080).
Western blot - Anti-RUNX3 antibody [EPR20680] (ab224642)

All lanes : Anti-RUNX3 antibody [EPR20680] (ab224642) at 1/1000 dilution

Lane 1 : Human thymus lysate
Lane 2 : Human tonsil lysate

Lysates/proteins at 20 µg per lane.

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

Developed using the ECL technique.

Predicted band size: 44, 46 kDa
Observed band size: 44,46 kDa why is the actual band size different from the predicted?

Exposure time : Lane 1: 3 minutes; Lane 2: 15 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.

This target detects both predicted isoforms 44 kDa and 46 kDa, consistent with what has been described in the literature (PMID: 23700080).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20680] (ab224642)

Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling RUNX3 with ab224642 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on lymphoid cells of human stomach (PMID:15514019; PMID:21786422) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20680] (ab224642)

Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling RUNX3 with ab224642 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on lymphoid cells of human gastric cancer (PMID:27566570). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20680] (ab224642)

Immunohistochemical analysis of paraffin-embedded human diffuse large B cell lymphoma tissue labeling RUNX3 with ab224642 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on human diffuse large B cell lymphoma (PMID:27184221). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-RUNX3 antibody [EPR20680] (ab224642)

RUNX3 was immunoprecipitated from 0.35 mg of Raji (human Burkitt's lymphoma cell line) whole cell lysate with ab224642 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224642 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: Raji whole cell lysate 10 µg (Input).

Lane 2: ab224642 IP in Raji whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab224642 in Raji whole cell lysate.

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

Exposure time: 10 second.

The molecular weight observed is consistent with what has been described in the literature (PMID:23700080).
Anti-RUNX3 antibody [EPR20680] (ab224642)

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