Anti-Histone H3 (citrulline R2 + R8 + R17) 抗体

• WB

Lab

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (AB5103)

Loading Control : GAPDH

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab5103 overnight at 4°C. Antibody binding was detected using Goat anti Rabbit IR680 secondary at a 1 : 10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

All lanes:

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103) at 0.2 µg/mL

Lane 1:

HL60 whole cell lysate (negative control) at 20 µg

Lane 2:

HL60 whole cell lysate + DMSO (solvent control) at 20 µg

Lane 3:

HL60 whole cell lysate + DMSO + Calcium Ionophore (positive control) at 20 µg

Secondary

All lanes:

Goat anti Rabbit IR680 at 1/10000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (AB5103)

This immunoprecipitation image is a comparison between ab5103 and the alternative recombinant multiclonal antibody ab281584.

Left side - Recombinant multiclonal to Histone H3 (citrulline R2 + R8 + R17) - ab281584
All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] ab281584 at 1/1000 dilution.
Lane 1 : HEK-293T (human embryonic kidney) transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours, whole cell lysate 10 μg.
Lane 2 : ab281584 IP in HEK-293T transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab281584 in HEK-293T transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours whole cell lysate

Right side : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)
All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody ab5103 at 1/1000 dilution.
Lane 1 : HEK-293T (human embryonic kidney) transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours, whole cell lysate 10 μg.
Lane 2 : ab5103 IP in HEK-293T transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab5103 in HEK-293T transfected with PADI4 expression vector containing a GFP-tag treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours whole cell lysate

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Predicted band size : 15kDa

Why choose a recombinant antibody?
Research with confidence – consistent and reproducible results with every batch
Long-term and scalable supply – powered by recombinant technology for fast production
Success from the first experiment – confirmed specificity through extensive validation
Ethical standards compliant – production is animal-free

All lanes:

Immunoprecipitation - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)

Predicted band size: 15 kDa

false

• WB

Lab

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (AB5103)

Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above .

Blots were developled with Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody

All lanes:

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103) at 1 µg/mL

Lane 1:

HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Ionophore treated Whole Cell Lysate at 10 µg with with 5% BSA

Lane 2:

HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Ionophore treated Whole Cell Lysate at 10 µg with with 5% milk

Lane 3:

HL60 (Human Caucasian promyelocytic leukaemia) DMSO and Calcium Ionophore treated Whole Cell Lysate at 10 µg with with 3% milk

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

true

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (AB5103)

This western blot image is a comparison between ab5103 and the alternative recombinant multiclonal antibody ab281584.

Left side - Recombinant multiclonal to Histone H3 (citrulline R2 + R8 + R17) - ab281584
All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] ab281584 at 1/1000 dilution.
Lane 1 : Untreated HEK-293T (human embryonic kidney) transfected with PADI4 expression vector containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293T transfected with PADI4 expression vector containing a myc-His-tag® treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours, whole cell lysate
Lane 3 : Untreated NIH/3T3 (mouse embryonic fibroblast) transfected with PADI4 expression vector containing a myc-His-tag®, whole cell lysate
Lane 4 : NIH/3T3 transfected with PADI4 expression vector containing a myc-His-tag® treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours, whole cell lysate

Right side : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)
All lanes : Anti-Histone H3 (citrulline R2 + R8 + R17) antibody ab5103 at 1/1000 dilution.
Lane 1 : Untreated HEK-293T (human embryonic kidney) transfected with PADI4 expression vector containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293T transfected with PADI4 expression vector containing a myc-His-tag® treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours, whole cell lysate
Lane 3 : Untreated NIH/3T3 (mouse embryonic fibroblast) transfected with PADI4 expression vector containing a myc-His-tag®, whole cell lysate
Lane 4 : NIH/3T3 transfected with PADI4 expression vector containing a myc-His-tag® treated with 10 mM calcium chloride and 10 μM lonomycin for 4 hours, 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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Why choose a recombinant antibody?
Research with confidence – consistent and reproducible results with every batch
Long-term and scalable supply – powered by recombinant technology for fast production
Success from the first experiment – confirmed specificity through extensive validation
Ethical standards compliant – production is animal-free

All lanes:

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103)

Predicted band size: 15 kDa

false

• WB

Unknown

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (AB5103)

Blocking buffer : 2% BSA

Gel type : MES

All lanes:

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103) at 1 µg/mL

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

Rat brain tissue lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

Exposure time: 1min

• WB

Unknown

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (AB5103)

Blocking buffer : 2% BSA

Gel type : MES

All lanes:

Western blot - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (ab5103) at 1 µg/mL

Lane 1:

NIH/3T3 (Mouse embryo fibroblast cell line) nuclear lysate at 10 µg

Lane 2:

PC12 (Rat adrenal pheochromocytoma cell line) nuclear lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

Exposure time: 1min

• PepArr

Lab

Peptide Array - Anti-Histone H3 (citrulline R2 + R8 + R17) antibody (AB5103)

All batches of ab5103 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - citrulline 2 + 8 + 17 peptide (ab32876), indicating that this antibody specifically recognises the Histone H3 - citrulline 2 + 8 + 17 modifications.

ab32876 - Histone H3 - citrulline 2 + 8 + 17

ab17566 - Histone H3 - unmodified