WB
Western blot analysis of SHMT2 / SHMT on different lysates with Rabbit anti-SHMT2 / SHMT antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate, Lane 2: Saos-2 cell lysate, Lane 3: HepG2 cell lysate, Lane 4: LNCaP cell lysate, Lane 5: Jurkat cell lysate, Lane 6: COS-1 cell lysate, Lane 7: Hepa1-6 cell lysate, Lane 8: MC/9 cell lysate, Lane 9: Mouse liver tissue lysate, Lane 10: Mouse kidney tissue lysate, Lane 11: Mouse skeletal muscle tissue lysate (low expression), Lane 12: Rat liver tissue lysate, Lane 13: Rat kidney tissue lysate, Lane 14: Rat skeletal muscle tissue lysate (low expression) Lysates/proteins at 20 µg/Lane. Exposure time: 3 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 at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-SHMT2 / SHMT antibody at 1/1,000 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 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-SHMT2 / SHMT antibody at 1/1,000 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 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.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-SHMT2 / SHMT antibody at 1/1,000 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 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.ICC/IF
Immunocytochemistry analysis of Hepa1-6 cells labeling SHMT2 / SHMT with Rabbit anti-SHMT2 / SHMT antibody at 1/200 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-SHMT2 / SHMT antibody at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.FC
Flow cytometric analysis of Hepa1-6 cells labeling SHMT2 / SHMT. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
SHMT2 / SHMT was immunoprecipitated from 0.2 mg LNCaP cell lysate with Rabbit anti-SHMT2 / SHMT antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-SHMT2 / SHMT antibody 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: LNCaP cell lysate (input), Lane 2: Rabbit anti-SHMT2 / SHMT antibody IP in LNCaP cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-SHMT2 / SHMT antibody in LNCaP cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 2 seconds.| Product Name | SHMT2 / SHMT Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human SHMT2 aa 1-504. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000 IHC:1:1000 ICC/IF:1:200-1:500 FC:1:500-1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | SHMT2 |
|---|---|
| Gene Synonyms | GLYA SHMT mSHMT NEDCASB HEL-S-51e |
| Gene Full Name | serine hydroxymethyltransferase 2 |
| Gene Summary | This gene encodes the mitochondrial form of a pyridoxal phosphate-dependent enzyme that catalyzes the reversible reaction of serine and tetrahydrofolate to glycine and 5,10-methylene tetrahydrofolate. The encoded product is primarily responsible for glycine synthesis. The activity of the encoded protein has been suggested to be the primary source of intracellular glycine. The gene which encodes the cytosolic form of this enzyme is located on chromosome 17. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Oct 2009] |
| Alternative Names | Serine hydroxymethyltransferase mitochondrial EC:2.1.2.1 SHMT Glycine hydroxymethyltransferase Serine methylase SHMT2 |
| Molecular Weight(MW) | 56kDa |
| Cellular Localization | Mitochondrion, Mitochondrion matrix, mitochondrion nucleoid, Mitochondrion inner membrane, Cytoplasm, Nucleus. |
WB
Western blot analysis of SHMT2 / SHMT on different lysates with Rabbit anti-SHMT2 / SHMT antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate, Lane 2: Saos-2 cell lysate, Lane 3: HepG2 cell lysate, Lane 4: LNCaP cell lysate, Lane 5: Jurkat cell lysate, Lane 6: COS-1 cell lysate, Lane 7: Hepa1-6 cell lysate, Lane 8: MC/9 cell lysate, Lane 9: Mouse liver tissue lysate, Lane 10: Mouse kidney tissue lysate, Lane 11: Mouse skeletal muscle tissue lysate (low expression), Lane 12: Rat liver tissue lysate, Lane 13: Rat kidney tissue lysate, Lane 14: Rat skeletal muscle tissue lysate (low expression) Lysates/proteins at 20 µg/Lane. Exposure time: 3 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 at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-SHMT2 / SHMT antibody at 1/1,000 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-SHMT2 / SHMT antibody at 1/1,000 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-SHMT2 / SHMT antibody at 1/1,000 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 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.
ICC/IF
Immunocytochemistry analysis of Hepa1-6 cells labeling SHMT2 / SHMT with Rabbit anti-SHMT2 / SHMT antibody at 1/200 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-SHMT2 / SHMT antibody at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.
FC
Flow cytometric analysis of Hepa1-6 cells labeling SHMT2 / SHMT. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
SHMT2 / SHMT was immunoprecipitated from 0.2 mg LNCaP cell lysate with Rabbit anti-SHMT2 / SHMT antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-SHMT2 / SHMT antibody 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: LNCaP cell lysate (input), Lane 2: Rabbit anti-SHMT2 / SHMT antibody IP in LNCaP cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-SHMT2 / SHMT antibody in LNCaP cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 2 seconds.| Application Notes | WB:1:5000 IHC:1:1000 ICC/IF:1:200-1:500 FC:1:500-1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
