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-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20358-120] to Histone H3 (citrulline R17)
Suitable for: ELISA, WB, Flow Cyt, IP, ICC/IF
Reacts with: Mouse, Rat, Human

Product name

Anti-Histone H3 (citrulline R17) antibody [EPR20358-120]
See all Histone H3 primary antibodies
Description

Rabbit monoclonal [EPR20358-120] to Histone H3 (citrulline R17)
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IP	Human
WB	Mouse Rat Recombinant fragment

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293T and C6 transfected with PADI4, then treated with CaCl2 and Ionomycin (ab120116) whole cell lysates; NIH/3T3 transfected with PADI4 then treated with CaCl2 whole cell lysate; E12 rat and mouse embryo lysates. ICC/IF: HEK-293T cells transfected with GFP-tagged PADI4, then treated with CaCl2. Flow Cyt: HEK-293T transfected with PADI4, then treated with CaCl2 and Ionomycin (ab120116). IP: HEK-293T transfected with PADI4 then treated with CaCl2 and Ionomycin (ab120116) 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, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20358-120
Isotype

IgG
Research areas

Western blot - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)

All lanes : Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407) at 1/5000 dilution

Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney) transfected with empty vector with GFP tag (vector control), then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours, whole cell lysate
Lane 2 : HEK-293T (Human epithelial cell line from embryonic kidney) transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours, whole cell lysate

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: 15 kDa

Exposure time: 3 seconds

Blocking/Dilution buffer: 5% BSA/TBST.

Histone H3R17 is citrullinated by PADI4 and CaCl₂ is used as a cofactor according to the literature (PMID: 16567635). Ionomycin is used to improve the modification by PADI4 according to the literature (PMID: 26360112).
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney) cells transfected with GFP only or a GFP-tagged PADI4 expression construct, then treated with 10mM CaCl₂ for 2 hours, labeling Histone H3 (citrulline R17) with ab219407 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing positive staining on HEK-293T cells transfected with a GFP-tagged PADI4 expression construct, then treated with 10mM CaCl₂ for 2 hours.

The nuclear counterstain is DAPI (blue).

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

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Flow Cytometry - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)

Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293T (Human epithelial cell line from embryonic kidney) transfected with empty vector (left panel) or PADI4 (WT, right panel), then treated with 10mM CaCl₂ and 10μM Ionomycin (ab120116) for 2 hours, labeling Histone H3 (citrulline R17) with ab219407 at 1/500 dilution, followed by Goat anti rabbit IgG (Alexa Fluor^® 647) at 1/2000 dilution.

Positive signal is obtained from HEK-293T cells transfected with WT PADI4 treated with 10mM CaCl₂ and 10μM Ionomycin (ab120116) for 2 hours.
Immunoprecipitation - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)

Histone H3 (citrulline R17) was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney) transfected with PADI4 (WT), then treated with 10mM CaCl₂ and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate with ab219407 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab219407 at 1/1000 dilution.

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

Lane 1: HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl₂ and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate 10 µg (Input).
Lane 2: ab219407 IP in HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl₂ and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219407 in HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl₂ and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)

All lanes : Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407) at 1/5000 dilution

Lane 1 : NIH/3T3 (Mouse embryonic fibroblast cell line) transfected with empty vector with GFP tag (vector control), then treated with 10mM CaCl2 for 2 hours, whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) transfected with PADI4 (WT) then treated with 10mM CaCl2 for 2 hours, whole cell lysate
Lane 3 : C6 (Rat glial tumor cell line) transfected with empty vector with GFP tag (vector control) then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours, whole cell lysate
Lane 4 : C6 (Rat glial tumor cell line) transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours, whole cell lysate

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: 15 kDa

Exposure time: 1 second

Blocking/Dilution buffer: 5% BSA/TBST.
Western blot - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)

Lane 1 : Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407) at 1/1000 dilution
Lane 2 : Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407) at 1/200 dilution

Lane 1 : E12 mouse embryo lysate
Lane 2 : E12 rat embryo lysate

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: 15 kDa

Blocking/Dilution buffer: 5% BSA/TBST.

Exposure time: Lane 1: 1 second; Lane 2: 3 minutes.
ELISA - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)

Direct ELISA using ab219407 at 0-1000 ng/ml, followed by Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at 1/2500 dilution.

Antigen concentration: 1000 ng/ml.
Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)

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