Overview

  • Product name
  • Description
    Mouse monoclonal to StAR
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, ELISA, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment: EAMQKALGIL SNQEGWKKES QQDNGDKVMS KVVPDVGKVF RLEVVVDQPM ERLYEELVER MEAMGEWNPN VKEIKVLQKI GKDTFITHEL AAEAAGNLVG , corresponding to amino acids 81-180 of Human StAR

Applications

Our Abpromise guarantee covers the use of ab58013 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
WB Use a concentration of 1 - 5 µg/ml.
ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. PubMed: 21094146

Target

  • Function
    Plays a key role in steroid hormone synthesis by enhancing the metabolism of cholesterol into pregnenolone. Mediates the transfer of cholesterol from the outer mitochondrial membrane to the inner mitochondrial membrane where it is cleaved to pregnenolone.
  • Tissue specificity
    Expressed in gonads, adrenal cortex and kidney.
  • Pathway
    Steroid metabolism; cholesterol metabolism.
  • Involvement in disease
    Defects in STAR are the cause of adrenal hyperplasia type 1 (AH1) [MIM:201710]. The most severe form of adrenal hyperplasia. It is a condition characterized by onset of profound adrenocortical insufficiency shortly after birth, hyperpigmentation reflecting increased production of pro-opiomelanocortin, elevated plasma renin activity as a consequence of reduced aldosterone synthesis, and male pseudohermaphroditism resulting from deficient fetal testicular testosterone synthesis. Affected individuals are phenotypic females irrespective of gonadal sex, and frequently die in infancy if mineralocorticoid and glucocorticoid replacement are not instituted.
  • Sequence similarities
    Contains 1 START domain.
  • Cellular localization
    Mitochondrion.
  • Information by UniProt
  • Database links
  • Alternative names
    • Cholesterol trafficker antibody
    • Luteinizing hormone induced protein antibody
    • mitochondrial antibody
    • Mitochondrial steroid acute regulatory protein antibody
    • StAR antibody
    • StAR related lipid transfer (START) domain containing 1 antibody
    • STAR_HUMAN antibody
    • StARD1 antibody
    • START domain containing 1 antibody
    • START domain containing protein 1 antibody
    • START domain-containing protein 1 antibody
    • Steroid acute regulatory protein antibody
    • Steroidogenic acute regulator antibody
    • Steroidogenic acute regulatory protein antibody
    • Steroidogenic acute regulatory protein mitochondrial antibody
    see all

Images

  • Anti-StAR antibody (ab58013) at 1/1000 dilution + Mouse liver lysates at 50 µg

    Secondary
    HRP-conjugated Goat polyclonal to mouse IgG at 1/2500 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 27,30 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 minutes


    Liver lysates prepared from mouse injected with StarD1 adenovirus expressing recombinant StarD1 protein.

    Primary diluted in TBST + 2.5% milk and incubated with sample for 12 hours at 4°C.

  • Anti-StAR antibody (ab58013) at 1/1000 dilution (in TBST + 2.5% milk for 16 hours at 4°C) + Whole cell lysate of human THP1 cells at 50 µg

    Secondary
    An HRP-conjugated Goat anti-mouse IgG polyclonal at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 30 kDa why is the actual band size different from the predicted?


    Exposure time: 2 minutes


    Blocking Step: 5% Milk for 4 hours at 22°C

    See Abreview

References

This product has been referenced in:
  • Meimaridou E  et al. NNT is a key regulator of adrenal redox homeostasis and steroidogenesis in male mice. J Endocrinol 236:13-28 (2018). Read more (PubMed: 29046340) »
  • Hou X  et al. Pubertal chlorocholine chloride exposure inhibits testicular testosterone synthesis by down-regulating steroidogenic enzymes in adult rats. Toxicol Lett 288:17-24 (2018). WB ; Rat . Read more (PubMed: 29447956) »
See all 10 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary rat cardiac fibroblasts)
Permeabilization
Yes - 1% Triton, 4 minutes
Specification
Primary rat cardiac fibroblasts
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 20 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human adrenal corticocarcinoma H295R cells)
Permeabilization
Yes - 0.5% TX100
Specification
Human adrenal corticocarcinoma H295R cells
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Jun 27 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Human adrenal corticocarcinoma H295R cells)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
25 µg
Specification
Human adrenal corticocarcinoma H295R cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Jun 27 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (mouse testis)
Permeabilization
Yes - 0.5% TX100
Specification
mouse testis
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Dec 02 2016

Answer

Thank you for contacting us.

This is not a phopspho specific antibody which means this antibody would detect total StAR protein i.e. phospho as well as non-phospho form. However it will not be able to distinguish between the two forms.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

ab58013 StAR antibody 4) Description of the problem (high background, no band, wrong band size...) No band and wrong band 5) Sample preparation: Name of the tissue/cell line: MLTC-1 Sample species: mouse Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysates Lysis buffer (Beyotime) RIPA lysis buffer Protease inhibitors: PMSF Phosphatase inhibitors NO Reducing agent NO Boiling for ≥5 min? yes/no YES Protein loaded ug/lane or cells/lane 80ug Positive control no Negative control no 6) Percentage of gel Type of membrane PVDF Protein transfer verified yes Blocking agent and concentration 5% nonfat dried milk Blocking time 2h Blocking temperature room temp 7) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution 1:1000 or 1:500 Diluent buffer 5% nonfat dried milk Incubation time >8h Incubation temperature: 4℃ 8) Secondary antibody: anti mouse Species: goat Reacts against: mouse Concentration or dilution 1:1000 Diluent buffer 5% nonfat dried milk Incubation time 2h Incubation temperature: room temp Fluorochrome or enzyme conjugate: no 9) Washing after primary and secondary antibodies: Buffer 1* PBST Number of washes 5 min every time, 4 times 10)Detection method:ALPHA INNOTECH(FLUROR CHEM FC2); acquire image 30 sec and 1 minute 11) How many times have you run this staining? 5 times Do you obtain the same results every time? yes What steps have you altered to try and optimize the use of this antibody? Change Concentration (1:1000, and 1:500)

Read More
Answer

Thank you for contacting Abcam regarding ab58013. I am sorry that you have been experiencing difficulties with this antibody in WB.  I have reviewed the protocol information provided and would like to make some suggestions to improve your results with this antibody. You indicated that you did not run the experiment under reducing conditions.  I would recommend adding either beta-mercaptoethanol or DTT to your samples.  Others who have obtained positive results have run the gel under reduced and denaturing conditions.  Additionally, I recommend using BSA instead of milk as a blocking reagent and antibody diluent.  Test this with the 1:1000 dilution, as 1:500 seems to result in higher background in your samples. I hope this information is helpful.  Please do not hesitate to contact us if you have any additional questions.

Read More

Answer

Indeed there are two entries in the Uniprot database, Q6IBK0 and P49675. Both are for the same protein. However the entry which is best to refer to is the P49675 as this is the entry which is manually annotated and curated (Swissprot). Indeed, Uniprot has entries from Swissprot (manually curated) and from TrEMBL (data automatically extrapolated from the EMBL database). We do have tested this antibody against StAR ab58013 in Western blot and on human samples and do thus guarantee this antibody in this application. I can therefore recommend this antibody for the use in Western blot on human cell line samples.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (THP-1 cells)
Loading amount
50 µg
Specification
THP-1 cells
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Sep 22 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (testis)
Loading amount
35 µg
Specification
testis
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 22°C

Abcam user community

Verified customer

Submitted Feb 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Liver)
Loading amount
50 µg
Specification
Liver
Gel Running Conditions
Reduced Denaturing (12%)
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 Nov 20 2009

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

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