Rabbit FH/Fumarase Polyclonal Antibody | anti-FH antibody

Anti-FH/Fumarase Picoband Antibody

Cat. Num. AAA1750744

• Gene Names: FH; MCL; FMRD; LRCC; HLRCC; MCUL1
• Reactivity: Human, Mouse, Rat; No cross reactivity with other proteins.
• Applications: Immunohistochemistry, Western Blot
• Purity: Immunogen affinity purified
• Synonyms: FH/Fumarase; Polyclonal Antibody; Anti-FH/Fumarase Picoband Antibody; Fumarate hydratase; mitochondrial; Fumarase; 4.2.1.2; FH; anti-FH antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat; No cross reactivity with other proteins.
• Clonality: Polyclonal
• Purity/Purification: Immunogen affinity purified
• Form/Format: Lyophilized
• Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml. (varies by lot)
• Sequence Length: 467

• Applicable Applications for anti-FH antibody: Immunohistochemistry (IHC), Western Blot (WB)
• Application Notes: WB: 0.1-0.5mug/ml; Immunohistochemistry (Paraffin-embedded Section): 0.5-1mug/ml
• Immunogen: A synthetic peptide corresponding to a sequence of human FH (YDKAAKIAKTAHKNGSTLKETAIELGYLTAEQFDEWVKPKDMLGPK).
• Subcellular Localization: Isoform Mitochondrial: Mitochondrion.
• Tissue Specificity: Expressed in red blood cells; underexpressed in red blood cells (cytoplasm) of patients with hereditary non- spherocytic hemolytic anemia of unknown etiology.
• Reconstitution: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Contents: Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
• Preparation and Storage: Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of FH using anti-FH antibody (MBS1750744).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat lymphaden tissue lysates,Lane 2: rat small intestine tissue lysates,Lane 3: rat gaster tissue lysates,Lane 4: mouse kidney tissue lysates,Lane 5: mouse testis tissue lysates,Lane 6: mouse gaster tissue lysates,Lane 7: human K562 whole cell lysates,Lane 8: human U937 whole cell lysates,Lane 9:human HL-60 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-FH antigen affinity purified polyclonal antibody at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for FH at approximately 49KD. The expected band size for FH is at 55KD. )

Immunohistochemistry (IHC)

(Figure 2. IHC analysis of FH using anti-FH antibody (MBS1750744).FH was detected in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-FH Antibody (MBS1750744) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )

Immunohistochemistry (IHC)

(Figure 3. IHC analysis of FH using anti-FH antibody (MBS1750744).FH was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-FH Antibody (MBS1750744) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )

Immunohistochemistry (IHC)

(Figure 4. IHC analysis of FH using anti-FH antibody (MBS1750744).FH was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-FH Antibody (MBS1750744) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )

Immunohistochemistry (IHC)

(Figure 5. IHC analysis of FH using anti-FH antibody (MBS1750744).FH was detected in paraffin-embedded section of mouse small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-FH Antibody (MBS1750744) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )

Immunohistochemistry (IHC)

(Figure 6. IHC analysis of FH using anti-FH antibody (MBS1750744).FH was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-FH Antibody (MBS1750744) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )

Related Product Information for anti-FH antibody

Description: Fumarase (or fumaratehydratase) is an enzyme that catalyzes the reversible hydration/dehydration of fumarate to malate. Fumarase comes in two forms: mitochondrial and cytosolic. The mitochondrial isoenzyme is involved in the Krebs Cycle (also known as the Tricarboxylic Acid Cycle [TCA] or the Citric Acid Cycle), and the cytosolic isoenzyme is involved in the metabolism of amino acids and fumarate. Subcellular localization is established by the presence of a signal sequence on the amino terminus in the mitochondrial form, while subcellular localization in the cytosolic form is established by the absence of the signal sequence found in the mitochondrial variety. This enzyme participates in 2 metabolic pathways: citric acid cycle, reductive citric acid cycle (CO2 fixation), and is also important in renal cell carcinoma. Mutations in this gene have been associated with the development of leiomyomas in the skin and uterus in combination with renal cell carcinoma.
Protein Function: Also acts as a tumor suppressor.

NCBI and Uniprot Product Information

• NCBI GI #: 19743875
• NCBI GeneID: 2271
• NCBI Accession #: NP_000134.2
• NCBI GenBank Nucleotide #: NM_000143.3
• UniProt Accession #: P07954; B1ANK7
• Molecular Weight: 50,213 Da
• NCBI Official Full Name: fumarate hydratase, mitochondrial
• NCBI Official Synonym Full Names: fumarate hydratase
• NCBI Official Symbol: FH
• NCBI Official Synonym Symbols: MCL; FMRD; LRCC; HLRCC; MCUL1
• NCBI Protein Information: fumarate hydratase, mitochondrial
• UniProt Protein Name: Fumarate hydratase, mitochondrial
• UniProt Gene Name: FH
• UniProt Synonym Gene Names: Fumarase

NCBI Description

The protein encoded by this gene is an enzymatic component of the tricarboxylic acid (TCA) cycle, or Krebs cycle, and catalyzes the formation of L-malate from fumarate. It exists in both a cytosolic form and an N-terminal extended form, differing only in the translation start site used. The N-terminal extended form is targeted to the mitochondrion, where the removal of the extension generates the same form as in the cytoplasm. It is similar to some thermostable class II fumarases and functions as a homotetramer. Mutations in this gene can cause fumarase deficiency and lead to progressive encephalopathy. [provided by RefSeq, Jul 2008]

Uniprot Description

Also acts as a tumor suppressor.MiscellaneousThere are 2 substrate-binding sites: the catalytic A site, and the non-catalytic B site that may play a role in the transfer of substrate or product between the active site and the solvent. Alternatively, the B site may bind allosteric effectors ().