Validation Data Gallery

Filter: AllWBIHCIF-PIF/ICCFC (Intra)Cytometric bead array

WB analysis using 66863-1-Ig (same clone as 66863-1-PBS)
Various lysates were subjected to SDS PAGE followed by western blot with 66863-1-Ig (Phospho-Histone H3 (Ser10) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control. This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
×
Various lysates were subjected to SDS PAGE followed by western blot with 66863-1-Ig (Phospho-Histone H3 (Ser10) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control. This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
WB analysis using 66863-1-Ig (same clone as 66863-1-PBS)
Non-treated and Calyculin A treated cells were subjected to SDS PAGE followed by western blot with 66863-1-Ig (Phospho-Histone H3 (Ser10) antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control. This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
×
Non-treated and Calyculin A treated cells were subjected to SDS PAGE followed by western blot with 66863-1-Ig (Phospho-Histone H3 (Ser10) antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control. This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IHC staining of Jurkat using 66863-1-Ig (same clone as 66863-1-PBS)
Immunohistochemical analysis of paraffin-embedded Jurkat cells slide using 66863-1-Ig (Phospho-Histone H3 (Ser10) antibody) at dilution of 1:4000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IHC staining of human renal cell carcinoma using 66863-1-Ig (same clone as 66863-1-PBS)
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma tissue slide using 66863-1-Ig (Phospho-Histone H3 (Ser10) antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IHC staining of mouse kidney using 66863-1-Ig (same clone as 66863-1-PBS)
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 66863-1-Ig (Phospho-Histone H3 (Ser10) antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IHC staining of human lung cancer using 66863-1-Ig (same clone as 66863-1-PBS)
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66863-1-Ig (Phospho-Histone H3 (Ser10) antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IF Staining of mouse testis using 66863-1-Ig (same clone as 66863-1-PBS)
Immunofluorescent analysis of (4% PFA) fixed mouse testis tissue using Phospho-Histone H3 (Ser10) antibody (66863-1-Ig, Clone: 4C7G2 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IF Staining of human breast cancer using 66863-1-Ig (same clone as 66863-1-PBS)
Immunofluorescent analysis of (4% PFA) fixed human breast cancer tissue using 66863-1-Ig (PHH3 antibody) at dilution of 1:100 and CoraLite488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IF Staining of C2C12 using 66863-1-Ig (same clone as 66863-1-PBS)
Immunofluorescent analysis of (4% PFA) fixed C2C12 cells using Phospho-Histone H3 (Ser10) antibody (66863-1-Ig, Clone: 4C7G2 ) at dilution of 1:1200 and CoraLite®594-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), Alpha Tubulin antibody (11224-1-AP, green). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IF Staining of HeLa using 66863-1-Ig (same clone as 66863-1-PBS)
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Phospho-Histone H3 (Ser10) antibody (66863-1-Ig, Clone: 4C7G2 ) at dilution of 1:1500 and CoraLite®594-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), Alpha Tubulin antibody (11224-1-AP, green). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IF Staining of MCF-7 using 66863-1-Ig (same clone as 66863-1-PBS)
Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using Phospho-Histone H3 (Ser10) antibody (66863-1-Ig, Clone: 4C7G2 ) at dilution of 1:1500 and CoraLite®594-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), Alpha Tubulin antibody (11224-1-AP, green). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
IF Staining of HeLa using 66863-1-Ig (same clone as 66863-1-PBS)
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Phospho-Histone H3 (Ser10) antibody (66863-1-Ig, Clone: 4C7G2 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1), Beta Tubulin antibody (80713-1-RR, Clone: 2O13, red). This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
FC experiment of HeLa using 66863-1-Ig (same clone as 66863-1-PBS)
1x10^6 nocodazole treated HeLa cells were intracellularly stained with 0.25 ug Phospho-Histone H3 (Ser10) Monoclonal antibody (66863-1-Ig, Clone:4C7G2) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1), and 0.25 ug Mouse IgG1 isotype control Mouse McAb (66360-1-Ig, Clone: 1F8D3). Cells were fixed with 4% PFA and permeabilized with 90% MeOH. This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
×
1x10^6 nocodazole treated HeLa cells were intracellularly stained with 0.25 ug Phospho-Histone H3 (Ser10) Monoclonal antibody (66863-1-Ig, Clone:4C7G2) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1), and 0.25 ug Mouse IgG1 isotype control Mouse McAb (66360-1-Ig, Clone: 1F8D3). Cells were fixed with 4% PFA and permeabilized with 90% MeOH. This data was developed using the same antibody clone with 66863-1-PBS in a different storage buffer formulation.
Cytometric bead array in cell lysate using MP50178-1
Cytometric bead array in cell lysate using MP50178-1, Phospho-Histone H3 (Ser10) Monoclonal Matched Antibody Pair, PBS Only. Capture antibody: 68345-1-PBS. Detection antibody: 66863-1-PBS. Cell lysate: Non-treated HeLa and Calyculin A treated HeLa (30μg/well). Non-related target OAT Monoclonal Matched Antibody Pair (MP50109-1P) was served as control.

Phospho-Histone H3 (Ser10) Monoclonal antibody, PBS Only (Detector)

Phospho-Histone H3 (Ser10) Monoclonal Antibody for WB, IHC, IF/ICC, IF-P, FC (Intra), Cytometric bead array, Indirect ELISA

Cat No. 66863-1-PBS

Clone No.4C7G2

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat, pig

Applications

WB, IHC, IF/ICC, IF-P, FC (Intra), Cytometric bead array, Indirect ELISA

Synonyms

Histone H3 (Ser10), p Histone H3, p Histone H3 (Ser10), p Histone H3 Ser10, p-Histone H3

Formulation: PBS Only

PBS and Azide

PBS Only

Conjugate: Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information

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MSDS

Tested Applications

Recommended dilution

Application	Dilution
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Product Information

66863-1-PBS targets Phospho-Histone H3 (Ser10) as part of a matched antibody pair:

MP50178-1: 68345-1-PBS capture and 66863-1-PBS detection (validated in Cytometric bead array)

Unconjugated mouse monoclonal antibody pair in PBS only (BSA and azide free) storage buffer at a concentration of 1 mg/mL, ready for conjugation.

This conjugation ready format makes antibodies ideal for use in many applications including: ELISAs, multiplex assays requiring matched pairs, mass cytometry, and multiplex imaging applications.Antibody use should be optimized by the end user for each application and assay.

Tested Reactivity	human, mouse, rat, pig
Host / Isotype	Mouse / IgG1
Class	Monoclonal
Type	Antibody
Immunogen	Peptide 相同性解析による交差性が予測される生物種
Full Name	histone cluster 1, H3a
Calculated molecular weight	15 kDa
Observed molecular weight	15-17 kDa
GenBank accession number	NM_003529
Gene Symbol	HIST1H3A
Gene ID (NCBI)	8350
RRID	AB_2882201
Conjugate	Unconjugated
Form	Liquid
Purification Method	Protein A purification
UNIPROT ID	P68431
Storage Buffer	PBS only , pH 7.3
Storage Conditions	Store at -80°C.

Background Information

Phospho-histone-H3 (PHH3) is a core histone protein, which in its phosphorylated state forms the principal constituents of eukaryotic chromatin, with histone H3 being phosphorylated at serine (Ser) 10 or Ser28 as well as its phosphorylation of Ser10 being strongly correlated with the late G2 to M-phase transition in mammalian mitotic cells. On the basis of previous research, a few cell line- and animal model-based researches have displayed an increase in phosphorylation of histone H3 at Ser10 (H3S10ph), the only histone marker that is involved in carcinogenesis and cellular transformation. Histone H3 phosphorylation on serine-10 is specific to mitosis and phosphorylated histone H3 (PHH3) proliferation markers (as counts defined per area or as indices defined per cell numbers) are increasingly being used to evaluate proliferation in various tumors.

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis using 66863-1-Ig (same clone as 66863-1-PBS)

WB analysis using 66863-1-Ig (same clone as 66863-1-PBS)

IHC staining of Jurkat using 66863-1-Ig (same clone as 66863-1-PBS)

IHC staining of human renal cell carcinoma using 66863-1-Ig (same clone as 66863-1-PBS)

IHC staining of mouse kidney using 66863-1-Ig (same clone as 66863-1-PBS)

IHC staining of human lung cancer using 66863-1-Ig (same clone as 66863-1-PBS)

IF Staining of mouse testis using 66863-1-Ig (same clone as 66863-1-PBS)

IF Staining of human breast cancer using 66863-1-Ig (same clone as 66863-1-PBS)

IF Staining of C2C12 using 66863-1-Ig (same clone as 66863-1-PBS)

IF Staining of HeLa using 66863-1-Ig (same clone as 66863-1-PBS)

IF Staining of MCF-7 using 66863-1-Ig (same clone as 66863-1-PBS)