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product-image-AAA19187_WB8.jpg WB (Western Blot) (Figure 8. Western blot analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Electrophoresis was performed on a 12% 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 brain tissue lysate,Lane 2: rat heart tissue lysate,Lane 3: rat kidney tissue lysate,Lane 4: rat testis tissue lysate,Lane 5: mouse brain tissue lysate,Lane 6: mouse heart tissue lysate,Lane 7: mouse kidney tissue lysate,Lane 8: mouse testis tissue lysate,Lane 9: mouse Neuro-2a whole cell lysate.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-Cytochrome C 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.)

Mouse Cytochrome C Monoclonal Antibody | anti-CYCS antibody

Anti-Cytochrome C Antibody (monoclonal, 15F10)

Gene Names
CYCS; CYC; HCS; THC4
Reactivity
Human, Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Immunocytochemistry, Flow Cytometry, Functional Assay
Purity
Immunogen Affinity Purified
Synonyms
Cytochrome C, Antibody; Anti-Cytochrome C Antibody (monoclonal, 15F10); Cytochrome c; CYCS; CYC; Cytochrome c, somatic; anti-CYCS antibody
Ordering
For Research Use Only!
Host
Mouse
Reactivity
Human, Mouse, Rat
Clonality
Monoclonal
Isotype
IgG1
Clone Number
15F10
Specificity
No cross reactivity with other proteins.
Purity/Purification
Immunogen Affinity Purified
Form/Format
Lyophilized
Sequence Length
105
Applicable Applications for anti-CYCS antibody
Western Blot (WB), Immunohistochemistry (IHC) Frozen/Paraffin, Immunocytochemistry (ICC), Flow Cytometry (FC/FACS)
Application Notes
WB: 0.1-0.5 mug/ml
IHC-P: 0.5-1 mug/ml
IHC-F: 0.5-1 mug/ml
ICC: 0.5-1 mug/ml
FC/FACS: 1-3ug/1x106 cells
Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections.
Immunogen
E Coli-derived human Cytochrome C recombinant protein (Position: G2-E105). Human Cytochrome C shares 91% amino acid (aa) sequence identity with both mouse and rat Cytochrome C.
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Relevant Detection Systems
It is recommended to use an Enhanced Chemiluminescent Kit with anti-Rabbit IgG for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit for IHC-P, IHC-F and ICC.
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.

WB (Western Blot)

(Figure 8. Western blot analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Electrophoresis was performed on a 12% 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 brain tissue lysate,Lane 2: rat heart tissue lysate,Lane 3: rat kidney tissue lysate,Lane 4: rat testis tissue lysate,Lane 5: mouse brain tissue lysate,Lane 6: mouse heart tissue lysate,Lane 7: mouse kidney tissue lysate,Lane 8: mouse testis tissue lysate,Lane 9: mouse Neuro-2a whole cell lysate.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-Cytochrome C 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.)

product-image-AAA19187_WB8.jpg WB (Western Blot) (Figure 8. Western blot analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Electrophoresis was performed on a 12% 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 brain tissue lysate,Lane 2: rat heart tissue lysate,Lane 3: rat kidney tissue lysate,Lane 4: rat testis tissue lysate,Lane 5: mouse brain tissue lysate,Lane 6: mouse heart tissue lysate,Lane 7: mouse kidney tissue lysate,Lane 8: mouse testis tissue lysate,Lane 9: mouse Neuro-2a whole cell lysate.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-Cytochrome C 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.)

WB (Western Blot)

(Figure 7. Western blot analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Electrophoresis was performed on a 12% 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: human Hela whole cell lysate,Lane 2: human HepG2 whole cell lysateLane 3: human K562 whole cell lysate,Lane 4: human Caco-2 whole cell lysate.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 mouse anti-Cytochrome C antigen affinity purified monoclonal 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-mouse 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.)

product-image-AAA19187_WB7.jpg WB (Western Blot) (Figure 7. Western blot analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Electrophoresis was performed on a 12% 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: human Hela whole cell lysate,Lane 2: human HepG2 whole cell lysateLane 3: human K562 whole cell lysate,Lane 4: human Caco-2 whole cell lysate.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 mouse anti-Cytochrome C antigen affinity purified monoclonal 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-mouse 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.)

IHC (Immunohistchemistry)

(Figure 6. IHC analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Cytochrome C 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 2ug/ml mouse anti-Cytochrome C Antibody (M03529-5) overnight at 4 degree C. Biotinylated goat anti-mouse 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.)

product-image-AAA19187_IHC6.jpg IHC (Immunohistchemistry) (Figure 6. IHC analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Cytochrome C 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 2ug/ml mouse anti-Cytochrome C Antibody (M03529-5) overnight at 4 degree C. Biotinylated goat anti-mouse 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.)

IHC (Immunohistochemistry)

(Figure 5. IHC analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Cytochrome C was detected in paraffin-embedded section of human intestinal 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 2ug/ml mouse anti-Cytochrome C Antibody (M03529-5) overnight at 4 degree C. Biotinylated goat anti-mouse 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.)

product-image-AAA19187_IHC5.jpg IHC (Immunohistochemistry) (Figure 5. IHC analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Cytochrome C was detected in paraffin-embedded section of human intestinal 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 2ug/ml mouse anti-Cytochrome C Antibody (M03529-5) overnight at 4 degree C. Biotinylated goat anti-mouse 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.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Cytochrome C 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 2ug/ml mouse anti-Cytochrome C Antibody (M03529-5) overnight at 4 degree C. Biotinylated goat anti-mouse 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.)

product-image-AAA19187_IHC4.jpg IHC (Immunohistochemistry) (Figure 4. IHC analysis of Cytochrome C using anti-Cytochrome C antibody (M03529-5).Cytochrome C 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 2ug/ml mouse anti-Cytochrome C Antibody (M03529-5) overnight at 4 degree C. Biotinylated goat anti-mouse 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.)

FCM (Flow Cytometry)

(Figure 3. Flow Cytometry analysis of K562 cells using anti-Cytochrome C antibody (M03529-5).Overlay histogram showing K562 cells stained with M03529-5 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cytochrome C Antibody (M03529-5, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

product-image-AAA19187_FCM3.jpg FCM (Flow Cytometry) (Figure 3. Flow Cytometry analysis of K562 cells using anti-Cytochrome C antibody (M03529-5).Overlay histogram showing K562 cells stained with M03529-5 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cytochrome C Antibody (M03529-5, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

FCM (Flow Cytometry)

(Figure 3. Flow Cytometry analysis of K562 cells using anti-Cytochrome C antibody (M03529-5).Overlay histogram showing K562 cells stained with M03529-5 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cytochrome C Antibody (M03529-5, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

product-image-AAA19187_FCM2.jpg FCM (Flow Cytometry) (Figure 3. Flow Cytometry analysis of K562 cells using anti-Cytochrome C antibody (M03529-5).Overlay histogram showing K562 cells stained with M03529-5 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cytochrome C Antibody (M03529-5, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

FCM (Flow Cytometry)

(Figure 1. Flow Cytometry analysis of A431 cells using anti-Cytochrome C antibody (M03529-5).Overlay histogram showing A431 cells stained with M03529-5 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cytochrome C Antibody (M03529-5, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight® 488 conjugated goat anti-mouse IgG (BA1126, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

product-image-AAA19187_FCM.jpg FCM (Flow Cytometry) (Figure 1. Flow Cytometry analysis of A431 cells using anti-Cytochrome C antibody (M03529-5).Overlay histogram showing A431 cells stained with M03529-5 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cytochrome C Antibody (M03529-5, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight® 488 conjugated goat anti-mouse IgG (BA1126, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
Related Product Information for anti-CYCS antibody
Description: Mouse IgG monoclonal antibody for Cytochrome C detection. Tested with WB, IHC-P, IHC-F, ICC, FCM in Human; Mouse; Rat.
Background: CYCS is also known as CYC, HCS or THC4. This gene encodes a small heme protein that functions as a central component of the electron transport chain in mitochondria. The encoded protein associates with the inner membrane of the mitochondrion where it accepts electrons from cytochrome b and transfers them to the cytochrome oxidase complex. This protein is also involved in initiation of apoptosis. Mutations in this gene are associated with autosomal dominant nonsyndromic thrombocytopenia. Numerous processed pseudogenes of this gene are found throughout the human genome.
References
1. García-Heredia JM, et al. Specific nitration of tyrosines 46 and 48 makes cytochrome c assemble a non-functional apoptosome. FEBS Lett, 2012 Jan 20. 2. Rajagopal BS, et al. The hydrogen-peroxide-induced radical behaviour in human cytochrome c-phospholipid complexes: implications for the enhanced pro-apoptotic activity of the G41S mutant. Biochem J, 2013 Dec 15. 3. Simon M, et al. Versatility of non-native forms of human cytochrome c: pH and micellar concentration dependence. J Biol Inorg Chem, 2013 Jan.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
11,749 Da
NCBI Official Full Name
cytochrome c
NCBI Official Synonym Full Names
cytochrome c, somatic
NCBI Official Symbol
CYCS
NCBI Official Synonym Symbols
CYC; HCS; THC4
NCBI Protein Information
cytochrome c
UniProt Protein Name
Cytochrome c
UniProt Gene Name
CYCS
UniProt Synonym Gene Names
CYC

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Product Notes

The CYCS cycs (Catalog #AAA19187) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-Cytochrome C Antibody (monoclonal, 15F10) reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's Cytochrome C can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC) Frozen/Paraffin, Immunocytochemistry (ICC), Flow Cytometry (FC/FACS). WB: 0.1-0.5 mug/ml IHC-P: 0.5-1 mug/ml IHC-F: 0.5-1 mug/ml ICC: 0.5-1 mug/ml FC/FACS: 1-3ug/1x106 cells Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. Researchers should empirically determine the suitability of the CYCS cycs for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Cytochrome C, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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