Antibodies, Proteins, Kits and Reagents for Life Science

284 784 ₸ Product size

100 µl 284 784 ₸

Order now and get it on Wednesday March 10, 2021

Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18876] to Histone H3 (ubiquityl K23)
Suitable for: Dot blot, IP, WB
Reacts with: Human, Recombinant fragment

Product name

Anti-Histone H3 (ubiquityl K23) antibody [EPR18876]
See all Histone H3 primary antibodies
Description

Rabbit monoclonal [EPR18876] to Histone H3 (ubiquityl K23)
Host species

Rabbit

Tested Applications & Species

Application	Species
Dot	Recombinant fragment
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Lysate of HeLa cells treated with 2mM thymidine for 18 hrs, followed by addition of fresh culture medium and incubation for 9 hrs, then treated with 2mM thymidine for 15 hrs, followed by addition of fresh culture medium and incubation for 4 hrs; Jurkat, HEK-293 lysates. IP: Lysate of HeLa cells treated with 2mM thymidine for 18 hrs, followed by addition of fresh culture medium and incubation for 9 hrs, then 2mM thymidine for 15 hrs, followed by addition of fresh culture medium and incubation for 4 hrs.
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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

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, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18876
Isotype

IgG
Research areas

Western blot - Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243)

All lanes : Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243) at 1/20000 dilution

Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Whole cell lysates from HeLa treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours. Cell were then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours

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: 15 kDa
Observed band size: 28 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression level of H3K23ubi is increased during S phase. PMID 24013172.

The MW is consistent with literature: PMID 24013172.
Immunoprecipitation - Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243)

Histone H3 (ubiquityl K23) was immunoprecipitated using ab194243 at 1/30 dilution from 0.35 mg of whole cell lysate from HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours, then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours.

Western blot was performed from the immunoprecipitate using ab194243 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: 10 µg of whole cell lysate from HeLa treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours, cells were then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours (Input).

Lane 2: ab194243 IP in whole cell lysate from HeLa treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours, cells were then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194243 in whole cell lysate from HeLa treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours, cells were then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours.

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

Exposure time: 3 seconds.

The band at around 35kDa can be explained as di-ubiquitinated K23 (PMID24013172).
Western blot - Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243)

All lanes : Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243) at 1/20000 dilution

Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate

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

Predicted band size: 15 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The MW is consistent with literature: PMID 24013172.
Dot Blot - Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243)

Dot blot analysis of Histone H3 (ubiquityl K23) labeled with ab194243 at 1/1000 dilution.

Lane 1: Histone H3 (ubiquitinated K23) peptide (aa20-29).

Lane 2: Histone H3 non- ubiquitinated peptide (aa22-31).

Lane 3: Histone H3 full length recombinant protein.

Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 was used as secondary antibody.

Blocking and diluting buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243)

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