MTHFD2 Recombinant Rabbit Monoclonal Antibody [PSH24-73]
cat.: HA724469
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: PSH24-73
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 38 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human MTHFD2 aa 51-100.
Positive control: HEK-293 (Human embryonic kidney cell)cell lysate, MDA-MB-231 (Human breast cancer cell)cell lysate, HeLa (Human cervical adenocarcinoma cell)cell lysate, 786-0 (Human renal clear cell adenocarcinoma cell)cell lysate, MEF (Mouse embryonic fibroblast)cell lysate, Mouse spleen tissue lysate, Mouse testis tissue lysate, Rat spleen tissue lysate, Rat testis tissue lysate.
Subcellular location: Mitochondrion.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:5,000
1:100
1:1,000
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P13995 Human | P18155 Mouse
Entrez Gene: 680308 Rat
Alternative names: Bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase, mitochondrial Descriptions Methenyltetrahydrofolate cyclohydrolase methylene tetrahydrofolate dehydrogenase (NAD+ dependent) methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 2 methylenetetrahydrofolate dehydrogenase (NADP+ dependent) methylenetetrahydrofolate dehydrogenase 2 MGC82516 MTDC_HUMAN MTHFD2 mthfd2 methylene tetrahydrofolate dehydrogenase (NAD+ dependent) NAD-dependent methylene tetrahydrofolate dehydrogenase NAD-dependent methylenetetrahydrofolate dehydrogenase NMDMC
Images
HA724469_1.jpg Fig1: Western blot analysis of MTHFD2 on different lysates with Rabbit anti-MTHFD2 antibody (HA724469) at 1/5,000 dilution.

Lane 1: HEK-293 (Human embryonic kidney cell)cell lysate
Lane 2: MDA-MB-231 (Human breast cancer cell)cell lysate
Lane 3: HeLa (Human cervical adenocarcinoma cell)cell lysate
Lane 4: 786-0 (Human renal clear cell adenocarcinoma cell)cell lysate
Lane 5: MEF (Mouse embryonic fibroblast)cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 8 seconds ; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724469, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 38 kDa
Observed band size: 35 kDa
HA724469_2.jpg Fig2: Western blot analysis of MTHFD2 on different lysates with Rabbit anti-MTHFD2 antibody (HA724469) at 1/5,000 dilution.

Lane 1: Mouse spleen tissue lysate
Lane 2: Mouse testis tissue lysate
Lane 3: Rat spleen tissue lysate
Lane 4: Rat testis tissue lysate

Lysates/proteins at 30 µg/Lane.
Exposure time: 8 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724469, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 38 kDa
Observed band size: 35 kDa
HA724469_3.jpg Fig3: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HEK-293 (Human embryonic kidney cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724469, 1/100, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA724469_4.jpg Fig4: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HEK-293 (Human embryonic kidney cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724469, 1/100, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: The mitochondria dyes was Mito-Tracker (Beyotime, Red). The nuclear counterstain was DAPI (Blue).
HA724469_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Tonsil
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724469_6.jpg Fig6: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Breast cancer
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724469_7.jpg Fig7: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Spleen
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724469_8.jpg Fig8: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Spleen
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724469_9.jpg Fig9: Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Spleen
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724469_10.jpg Fig10: Application: Flow Cytometry (Intra)

Species: Human
Sample: HEK-293 (Human embryonic kidney cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: HA724469(1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.

HA724469_11.jpg Fig11: Application: Flow Cytometry (Intra)

Species: Mouse
Sample: MEF (Mouse embryonic fibroblast)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: HA724469(1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.

HA724469_12.jpg Fig12: Immunoprecipitation (IP)

MTHFD2 was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA724469 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724469 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA724469 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724469 in HeLa cell lysate

Exposure time: 5 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724469, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 38 kDa
Observed band size: 35 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.