M1309-1

Histone H3 Mouse Monoclonal Antibody [A11-D7]

cat.: M1309-1

Product Type:	Mouse monoclonal IgG2b, primary antibodies
Species reactivity:	Human, Mouse, Rat, Zebrafish
Applications:	WB, IF-Cell, IHC-P, IF-Tissue
Clonality:	Monoclonal
Clone number:	A11-D7

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 15 kDa
Isotype:	IgG2b
Immunogen:	Synthetic peptide within N-terminal human Histone H3.
Positive control:	HeLa cell lysate, A549 cell lysate, HT-29 cell lysate, HEK-293 cell lysate, C2C12 cell lysate, L-929 cell lysate, C6 cell lysate, zebrafish tissue lysates, HepG2 cell lysate, A431 cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Hela, F9, human skin tissue, human liver tissue, human testis tissue, mouse brain tissue, mouse testis tissue, rat testis tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P IF-Tissue	1:5,000-1:10,000 1:200-1:500 1:1,000 1:200
Uniprot #:	SwissProt: P68431 Human \| P84243 Human \| Q16695 Human \| Q6NXT2 Human \| Q71DI3 Human \| P68433 Mouse \| P84228 Mouse \| Q6LED0 Rat
Alternative names:	H3 histone family member E pseudogene H3 histone family member A H3/A H31_HUMAN H3F3 H3FA Hist1h3a HIST1H3B HIST1H3C HIST1H3D HIST1H3E HIST1H3F HIST1H3G HIST1H3H HIST1H3I HIST1H3J HIST3H3 histone 1 H3a Histone cluster 1 H3a Histone H3 3 pseudogene 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

Images

Fig1: Western blot analysis of Histone H3 on different lysates with Mouse anti-Histone H3 antibody (M1309-1) at 1/10,000 dilution and competitor's antibody at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A549 cell lysate
Lane 3: HT-29 cell lysate
Lane 4: HEK-293 cell lysate
Lane 5: C2C12 cell lysate
Lane 6: L-929 cell lysate
Lane 7: C6 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 18 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (M1309-1) at 1/10,000 dilution and competitor's antibody at 1/5,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of Histone H3 on zebrafish tissue lysates with Mouse anti-Histone H3 antibody (M1309-1) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 1 minute;

10% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (M1309-1) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature.

	Fig3: Western blot analysis of Histone H3 on different lysates with Mouse anti-Histone H3 antibody (M1309-1) at 1/5,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: HEK-293 cell lysate (15 µg/Lane) Lane 3: HepG2 cell lysate (15 µg/Lane) Lane 4: A431 cell lysate (15 µg/Lane) Lane 5: MCF7 cell lysate (15 µg/Lane) Lane 6: NIH/3T3 cell lysate (15 µg/Lane) Lane 7: C2C12 cell lysate (15 µg/Lane) Lane 8: C6 cell lysate (15 µg/Lane) Lane 9: PC-12 cell lysate (15 µg/Lane) Predicted band size: 15 kDa Observed band size: 15 kDa Exposure time: 17 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (M1309-1) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:50,000 dilution was used for 1 hour at room temperature.
	Fig4: ICC staining of Histone H3 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (M1309-1, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig5: ICC staining of Histone H3 in F9 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (M1309-1, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

	Fig6: Immunohistochemical analysis of paraffin-embedded human skin tissue with Mouse anti-Histone H3 antibody (M1309-1) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1309-1) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-Histone H3 antibody (M1309-1) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1309-1) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig8: Immunohistochemical analysis of paraffin-embedded human testis tissue with Mouse anti-Histone H3 antibody (M1309-1) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1309-1) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig9: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Mouse anti-Histone H3 antibody (M1309-1) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1309-1) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig10: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Mouse anti-Histone H3 antibody (M1309-1) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1309-1) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig11: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Mouse anti-Histone H3 antibody (M1309-1) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1309-1) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.