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Cytochrome C Recombinant Rabbit Monoclonal Antibody [SC69-08]

Usd: 385

Specification

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Catalog# ET1610-60

Cytochrome C Recombinant Rabbit Monoclonal Antibody [SC69-08]

Application

  • WB

  • IF-Tissue

  • IHC-P

  • IP

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Cytochrome C Recombinant Rabbit Monoclonal Antibody [SC69-08]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human Cytochrome C aa 20-60.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Tissue, IHC-P, IP, IF-Cell

Molecular Weight

Predicted band size: 12 kDa

Positive Control

HeLa cell lysate, C6 cell lysate, Mouse brain tissue lysate, Mouse kidney tissue lysate, Rat brain tissue lysate, Rat kidney tissue lysate, Jurkat cell lysate, Human kidney tissue lysate, C6, human kidney tissue, mouse liver tissue, mouse kidney tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

SC69-08

RRID

AB_2940742

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Tissue

  • 1:50-1:100

  • IHC-P

  • 1:1,000-1:5,000

  • IP

  • Use at an assay dependent concentration.

  • IF-Cell

  • 1:100

Target

Function

Cytochrome c is a well characterized mobile electron transport protein that is essential to energy conversion in all aerobic organisms. In mammalian cells, this highly conserved protein is normally localized to the mitochondrial intermembrane space. More recent studies have identifed cytosolic cytochrome c as a factor necessary for activation of apoptosis. During apoptosis, cytochrome c is translocated from the mitochondrial membrane to the cytosol, where it is required for activation of caspase-3 (CPP32). Overexpression of Bcl-2 has been shown to prevent the translocation of cytochrome c, thereby blocking the apoptotic process. Overexpression of Bax has been shown to induce the release of cytochrome c and to induce cell death. The release of cytochrome c from the mitochondria is thought to trigger an apoptotic cascade, whereby Apaf-1 binds to Apaf-3 (caspase-9) in a cytochrome c-dependent manner, leading to caspase-9 cleavage of caspase-3.

Background References

1. Liu Z et al. Mark4 promotes oxidative stress and inflammation via binding to PPAR and activating NF-kB pathway in mice adipocytes. Sci Rep 6:21382 (2016).

2. Qi Y et al. PGC-1a Silencing Compounds the Perturbation of Mitochondrial Function Caused by Mutant SOD1 in Skeletal Muscle of ALS Mouse Model. Front Aging Neurosci 7:204 (2015).

Sequence Similarity

Belongs to the cytochrome c family.

Post-translational Modification

Binds 1 heme group per subunit.; Phosphorylation at Tyr-49 and Tyr-98 both reduce by half the turnover in the reaction with cytochrome c oxidase, down-regulating mitochondrial respiration.

Subcellular Location

Mitochondrion intermembrane space.

UNIPROT

SwissProt: P99999 Human

SwissProt: P62897 Mouse

SwissProt: P62898 Rat

Synonyms

CYC antibody

CYC_HUMAN antibody

CYCS antibody

Cytochrome c antibody

Cytochrome c somatic antibody

HCS antibody

THC4 antibody

Images

  • Western blot analysis of Cytochrome C on different lysates with Rabbit anti-Cytochrome C antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (10 µg/Lane)<br />Lane 2: C6 cell lysate (10 µg/Lane)<br />Lane 3: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 4: Mouse kidney tissue lysate (20 µg/Lane)<br />Lane 5: Rat brain tissue lysate (20 µg/Lane)<br />Lane 6: Rat kidney tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 12 kDa<br />Observed band size: 12 kDa<br /><br />Exposure time: 4 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Cytochrome C on different lysates with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (10 µg/Lane)
    Lane 2: C6 cell lysate (10 µg/Lane)
    Lane 3: Mouse brain tissue lysate (20 µg/Lane)
    Lane 4: Mouse kidney tissue lysate (20 µg/Lane)
    Lane 5: Rat brain tissue lysate (20 µg/Lane)
    Lane 6: Rat kidney tissue lysate (20 µg/Lane)

    Predicted band size: 12 kDa
    Observed band size: 12 kDa

    Exposure time: 4 seconds; ECL: K1801;

    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 (ET1610-60) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Cytochrome C on different lysates with Rabbit anti-Cytochrome C antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (15 µg/Lane)<br />Lane 2: Jurkat cell lysate (15 µg/Lane)<br />Lane 3: C6 cell lysate (15 µg/Lane)<br />Lane 4: Human kidney tissue lysate (20 µg/Lane)<br />Lane 5: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 6: Rat brain tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 12 kDa<br />Observed band size: 12 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Cytochrome C on different lysates with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (15 µg/Lane)
    Lane 2: Jurkat cell lysate (15 µg/Lane)
    Lane 3: C6 cell lysate (15 µg/Lane)
    Lane 4: Human kidney tissue lysate (20 µg/Lane)
    Lane 5: Mouse brain tissue lysate (20 µg/Lane)
    Lane 6: Rat brain tissue lysate (20 µg/Lane)

    Predicted band size: 12 kDa
    Observed band size: 12 kDa

    Exposure time: 3 minutes; ECL: K1801;
    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 (ET1610-60) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of C6 cells labeling Cytochrome C with Rabbit anti-Cytochrome C antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Cytochrome C antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of C6 cells labeling Cytochrome C with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Cytochrome C antibody (ET1610-60) 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 (M1305-2, 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.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Cytochrome C antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/5,000 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.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/5,000 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 (ET1610-60) at 1/5,000 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.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Cytochrome C antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/5,000 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.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/5,000 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 (ET1610-60) at 1/5,000 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.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Cytochrome C antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Cytochrome C antibody (ET1610-60) 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 (ET1610-60) 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.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Cytochrome C antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) 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.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Cytochrome C antibody (ET1610-60) 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 (ET1610-60) 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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