KMT3A / SETD2 Recombinant Rabbit Monoclonal Antibody [PSH20-36]
cat.: HA751752
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, IP
Clonality: Monoclonal
Clone number: PSH20-36
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 288 kDa
Isotype: IgG
Immunogen: Recombinant protein within human SETD2 aa 2,051-2,564.
Positive control: Jurkat cell lysate, Daudi cell lysate, HeLa cell lysate, MEF cell lysate, C6 cell lysate, PC-12 cell lysate, MEF, C6, human small intestine tissue, human testis tissue, mouse small intestine tissue, mouse testis tissue, rat small intestine tissue, rat testis tissue.
Subcellular location: Nucleus, Chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  IP
1:5,000
1:100
1:200
1-2μg/sample
Uniprot #: SwissProt: Q9BYW2 Human | E9Q5F9 Mouse
Entrez Gene: 316013 Rat
Alternative names: EC 2.1.1.43 FLJ16420 FLJ22472 FLJ23184 FLJ45883 HBP231 HIF 1 HIF-1 HIF1 HIP-1 Histone lysine N methyltransferase SETD2 Histone-lysine N-methyltransferase SETD2 hSET2 HSPC069 Huntingtin interacting protein 1 Huntingtin interacting protein Huntingtin interacting protein B Huntingtin interacting protein HYPB Huntingtin yeast partner B Huntingtin-binding protein, 231-KD Huntingtin-interacting protein 1 Huntingtin-interacting protein B HYPB KIAA1732 KMT3A Lysine N methyltransferase 3A Lysine N-methyltransferase 3A p231HBP SET domain containing 2 SET domain-containing protein 2 SET2 SETD2 SETD2_HUMAN
Images
HA751752_1.jpg Fig1: Western blot analysis of KMT3A / SETD2 on different lysates with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/5,000 dilution.

Lane 1: Jurkat cell lysate
Lane 2: Daudi cell lysate
Lane 3: HeLa cell lysate
Lane 4: MEF cell lysate
Lane 5: C6 cell lysate
Lane 6: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 288 kDa
Observed band size: 460 kDa

Exposure time: 30 seconds; ECL: K1801;

3-8% 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 (HA751752) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA751752_2.jpg Fig2: Immunocytochemistry analysis of MEF cells labeling KMT3A / SETD2 with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA751752_3.jpg Fig3: Immunocytochemistry analysis of C6 cells labeling KMT3A / SETD2 with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA751752_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751752) at 1/200 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.
HA751752_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751752) at 1/200 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.
HA751752_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751752) at 1/200 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.
HA751752_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751752) at 1/200 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.
HA751752_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751752) at 1/200 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.
HA751752_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-KMT3A / SETD2 antibody (HA751752) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751752) at 1/200 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.
HA751752_10.jpg Fig10: KMT3A / SETD2 was immunoprecipitated from 0.2 mg Daudi cell lysate with HA751752 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751752 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: Daudi cell lysate (input)
Lane 2: HA751752 IP in Daudi cell lysate
Lane 3: Rabbit IgG instead of HA751752 in Daudi cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 46 seconds; ECL: K1802
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.