Overview

  • Product name
    Anti-Citrate synthetase antibody
    See all Citrate synthetase primary antibodies
  • Description
    Rabbit polyclonal to Citrate synthetase
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IP, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
    Predicted to work with: Cow, Pig, Xenopus laevis
  • Immunogen

    Recombinant fragment within Human Citrate synthetase aa 44-316. The exact sequence is proprietary. NP_938083

  • Positive control
    • WB: HEK-293T, A431, HeLa and HepG2 whole cell lysate (ab7900) and zebrafish skeletal muscle tissue and mouse liver mitochondrial homogenates. ICC/IF: HeLa and U-2 OS cells. IHC-P: Human colon, mouse intestine, mouse muscle and NCI-N87 xenograft tissues. IP: HEK-293T whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab96600 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100 - 1/1000.
IP 1/100 - 1/500.
IHC-P 1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/500 - 1/3000. Predicted molecular weight: 52 kDa.

Target

Images

  • All lanes : Anti-Citrate synthetase antibody (ab96600) at 1/1000 dilution

    Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 52 kDa



    10% SDS-PAGE.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Citrate synthetase (green) with ab96600 at a dilution of 1/200.

    Cells were fixed in 4% paraformaldehyde at room temperature for 15 minutes. Alpha Tubulin (red) was stained with ab184613 at a dilution of 1/1000. Blue: Hoechst 33342 staining.

  • Anti-Citrate synthetase antibody (ab96600) at 1/1000 dilution + Rat muscle tissue extracts at 50 µg

    Predicted band size: 52 kDa



    10% SDS-PAGE.

  • Anti-Citrate synthetase antibody (ab96600) at 1/1000 dilution + Rat brain tissue extracts at 50 µg

    Secondary
    HRP-conjugated anti-rabbit IgG

    Predicted band size: 52 kDa



    10% SDS-PAGE.

  • Anti-Citrate synthetase antibody (ab96600) at 1/1000 dilution + Mouse brain tissue extracts at 50 µg

    Secondary
    HRP-conjugated anti-rabbit IgG

    Predicted band size: 52 kDa



    10% SDS-PAGE.

  • Immunocytochemistry/Immunofluorescence analysis of U-2 OS (Human bone osteosarcoma epithelial cell line) cells labelling Citrate synthetase (green) with ab96600 at a dilution of 1/500.

    Cells were fixed in 4% paraformaldehyde at room temperature for 15 minutes. Alpha Tubulin (red) was stained with ab184613 at a dilution of 1/1000. Blue: Hoechst 33342 staining.

  • IHC image of ab96600 staining in human colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab96600, 5 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunoprecipitation of Citrate synthetase from HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate using 5 μg of ab96600.

    Western blot analysis was performed using ab96600.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse intestine tissue labeling Citrate synthetase with ab96600 at a dilution of 1/500.

    Antigen Retrieval: EDTA based buffer, 15 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse muscle tissue labeling Citrate synthetase with ab96600 at a dilution of 1/500.

    Antigen Retrieval: Citrate buffer, pH 6.0, 15 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of NCI-N87 xenograft tissue labeling Citrate synthetase with ab96600 at a dilution of 1/500.

    Antigen Retrieval: EDTA based buffer, 15 mins.

  • All lanes : Anti-Citrate synthetase antibody (ab96600) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish liver homogenate (20ug)
    Lane 3 : Zebrafish skeletal muscle homogenate (20ug)
    Lane 4 : Mouse liver mitochondrial homogenate (20ug)

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 52 kDa
    Observed band size: 47 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 minutes

References

This product has been referenced in:
  • Jin F  et al. HIF-1a-induced miR-23a~27a~24 cluster promotes colorectal cancer progression via reprogramming metabolism. Cancer Lett 440-441:211-222 (2019). Read more (PubMed: 30393198) »
  • Fritzen AM  et al. Fatty acid type-specific regulation of SIRT1 does not affect insulin sensitivity in human skeletal muscle. FASEB J 33:5510-5519 (2019). Read more (PubMed: 30707625) »
See all 61 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HCT 116 Human colorectal carcinoma)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
HCT 116 Human colorectal carcinoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dr. Dimitra Kalamida

Verified customer

Submitted Mar 28 2019

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Bos taurus Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (4-15% gradient gel)
Loading amount
20 µg
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Julie Kim

Verified customer

Submitted Apr 17 2017

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Sample
Human Cell (MRC5 normal lung fibroblasts)
Specification
MRC5 normal lung fibroblasts
Permeabilization
Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT
Fixative
Paraformaldehyde

Dr. Dimitra Kalamida

Verified customer

Submitted Feb 13 2015

Application
Western blot
Loading amount
0.5 µg
Gel Running Conditions
Reduced Denaturing (4-15% Tris-glycine)
Sample
Pig Recombinant protein (Citrate Synthase from porcine heart (Sigma))
Specification
Citrate Synthase from porcine heart (Sigma)
Blocking step
Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Miss. Rachel Loh

Verified customer

Submitted Jan 15 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (10% 37.5:1)
Sample
Pig Tissue lysate - other (Skeletal muscle mitochondria purified)
Specification
Skeletal muscle mitochondria purified
Blocking step
3% Casein in PBS with 0.1% Tween-20 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 27 2014

Application
Western blot
Loading amount
60 µg
Gel Running Conditions
Reduced Denaturing (10% 37.5:1)
Sample
Cow Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
3% Casein in PBS with 0.1% Tween-20 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 27 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin)
Specification
skin
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 100mM sodium cytrate
Permeabilization
No
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Ms. Hadas Pahima

Verified customer

Submitted Oct 10 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (brain)
Loading amount
30 µg
Specification
brain
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Ilan Sela

Verified customer

Submitted Aug 07 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (C2C12)
Loading amount
20 µg
Specification
C2C12
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 26 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (L6 myotubes)
Loading amount
20 µg
Specification
L6 myotubes
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Feb 23 2011

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

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