Rabbit Recombinant Monoclonal H3 citrulline R17 antibody. Suitable for IP, ELISA, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat, Recombinant fragment - Human samples. Cited in 11 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | ELISA | WB | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|
Human | Tested | Expected | Tested | Tested | Tested |
Mouse | Expected | Expected | Tested | Expected | Expected |
Rat | Expected | Expected | Tested | Expected | Expected |
Recombinant fragment - Human | Not recommended | Tested | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes - |
Species Rat | Dilution info 1/1000 | Notes - |
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
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Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
H3FA, HIST1H3A, H3C2, H3FL, HIST1H3B, H3C3, H3FC HIST1H3C, H3C4, H3FB, HIST1H3D, H3C6, H3FD, HIST1H3E, H3C7, H3FI, HIST1H3F, H3C8, H3FH, HIST1H3G, H3C10, H3FK, HIST1H3H, H3C11, H3FF, HIST1H3I, H3C12, H3FJ, HIST1H3J, H3FC, HIST1H3C, H3C1, Histone H3.1, Histone H3/a, Histone H3/b, Histone H3/c, Histone H3/d, Histone H3/f, Histone H3/h, Histone H3/i, Histone H3/j, Histone H3/k, Histone H3/l, H3R17cit
Rabbit Recombinant Monoclonal H3 citrulline R17 antibody. Suitable for IP, ELISA, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat, Recombinant fragment - Human samples. Cited in 11 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR20358-120
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
ELISA analysis of Human Histone H3 (citrulline R17) recombinant protein at 100 ng/ml with ab219407. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
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) (Goat Anti-Rabbit IgG H&L (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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
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 (Ionomycin Ca2+ Salt, Ca2+ ionophore 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) (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 (Ionomycin Ca2+ Salt, Ca2+ ionophore 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 (Ionomycin Ca2+ Salt, Ca2+ ionophore ab120116) for 2 hours, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab219407 in HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin (Ionomycin Ca2+ Salt, Ca2+ ionophore ab120116) for 2 hours, whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407)
Predicted band size: 15 kDa
Exposure time: 1s
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).
All lanes: Western blot - 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 (Ionomycin Ca2+ Salt, Ca2+ ionophore ab120116) for 2 hour at 10 µg
Lane 2: HEK-293T (Human epithelial cell line from embryonic kidney) transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10µM Ionomycin (Ionomycin Ca2+ Salt, Ca2+ ionophore ab120116) for 2 hours, whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 15 kDa
Observed band size: 15 kDa
Exposure time: 3s
Blocking/Dilution buffer: 5% BSA/TBST.
Exposure time: Lane 1: 1 second; Lane 2: 3 minutes.
Lane 1: Western blot - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407) at 1/1000 dilution
Lane 2: Western blot - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407) at 1/200 dilution
Lane 1: E12 mouse embryo lysate at 10 µg
Lane 2: E12 rat embryo lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (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.
All lanes: Western blot - 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 at 10 µg
Lane 2: NIH/3T3 (Mouse embryonic fibroblast cell line) transfected with PADI4 (WT) then treated with 10mM CaCl2 for 2 hours, whole cell lysate at 10 µg
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 (Ionomycin Ca2+ Salt, Ca2+ ionophore ab120116) for 2 hours, whole cell lysate at 10 µg
Lane 4: C6 (Rat glial tumor cell line) transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10µM Ionomycin (Ionomycin Ca2+ Salt, Ca2+ ionophore ab120116) for 2 hours, whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 15 kDa
Observed band size: 15 kDa
Exposure time: 1s
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.
Intracellular 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 (Ionomycin Ca2+ Salt, Ca2+ ionophore 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 (Ionomycin Ca2+ Salt, Ca2+ ionophore ab120116) for 2 hours.
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