Overview

  • Product name

    Anti-Steroidogenic Factor 1/SF-1 antibody
    See all Steroidogenic Factor 1/SF-1 primary antibodies
  • Description

    Rabbit polyclonal to Steroidogenic Factor 1/SF-1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Horse, Cow, Pig
  • Immunogen

    Synthetic peptide corresponding to Human Steroidogenic Factor 1/SF-1 aa 400 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab74765, ab74766)

  • Positive control

    • Recombinant Human Steroidogenic Factor 1/SF-1 protein (ab114599) can be used as a positive control in WB. This antibody gave a positive signal in the following human tissue lysates: Testis; Ovary; Thymus.
  • General notes

    Previously labelled as Steroidogenic Factor 1. 

Properties

Applications

Our Abpromise guarantee covers the use of ab65815 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 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 52 kDa).
IHC-P Use at an assay dependent concentration.

Target

  • Function

    Transcriptional activator. Seems to be essential for sexual differentiation and formation of the primary steroidogenic tissues. Binds to the Ad4 site found in the promoter region of steroidogenic P450 genes such as CYP11A, CYP11B and CYP21B. Also regulates the AMH/Muellerian inhibiting substance gene as well as the AHCH and STAR genes. 5'-YCAAGGYC-3' and 5'-RRAGGTCA-3' are the consensus sequences for the recognition by NR5A1. The SFPQ-NONO-NR5A1 complex binds to the CYP17 promoter and regulates basal and cAMP-dependent transcriptional avtivity. Binds phosphatidylcholine (By similarity). Binds phospholipids with a phosphatidylinositol (PI) headgroup, in particular PI(3,4)P2 and PI(3,4,5)P3.
  • Involvement in disease

    Defects in NR5A1 are a cause of 46,XY disorder of sex development (46,XY DSD) [MIM:612965]; also known as XY sex reversal with or without adrenal failure. A congenital condition in which development of chromosomal, gonadal, or antomic sex is atypical. 46,XY DSD is a disorder of gonadal (testicular) development, which may be complete or partial. The complete form includes streak gonads, normal mullerian structures, and normal female external genitalia. The partial form includes ambiguous external genitalia and partial development of mullerian and wolffian structures.
    Defects in NR5A1 are a cause of adrenocortical insufficiency without ovarian defect (ACIWOD) [MIM:184757]. ACIWOD is characterized by severe 'slackness' muscular hypotonia. There is decreased sodium, increased potassium and elevated ACTH.
    Defects in NR5A1 are the cause of premature ovarian failure type 7 (POF7) [MIM:612964]. An ovarian disorder defined as the cessation of ovarian function under the age of 40 years. It is characterized by oligomenorrhea or amenorrhea, in the presence of elevated levels of serum gonadotropins and low estradiol.
  • Sequence similarities

    Belongs to the nuclear hormone receptor family. NR5 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • Post-translational
    modifications

    Acetylation stimulates the transcriptional activity.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • AD4BP antibody
    • Adrenal 4 binding protein antibody
    • Adrenal 4-binding protein antibody
    • ELP antibody
    • FTZ 1 antibody
    • FTZ1 antibody
    • FTZF 1 antibody
    • FTZF1 antibody
    • Fushi tarazu factor (Drosophila) homolog 1 antibody
    • Fushi tarazu factor homolog 1 antibody
    • hSF 1 antibody
    • NR5A1 antibody
    • Nuclear receptor AdBP4 antibody
    • Nuclear receptor subfamily 5 group A member 1 antibody
    • POF7 antibody
    • SF 1 antibody
    • SF-1 antibody
    • SF1 antibody
    • SPGF8 antibody
    • SRXY3 antibody
    • Steroid hormone receptor Ad4BP antibody
    • Steroidogenic factor 1 antibody
    • Steroidogenic factor 1 nuclear receptor antibody
    • STF 1 antibody
    • STF-1 antibody
    • STF1 antibody
    • STF1_HUMAN antibody
    see all

Images

  • All lanes : Anti-Steroidogenic Factor 1/SF-1 antibody (ab65815) at 1 µg/ml

    Lane 1 : Human testis tissue lysate - total protein
    Lane 2 : Human ovary tissue lysate - total protein

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 52 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 30 kDa (possible non-specific binding), 70 kDa (possible non-specific binding)


    Exposure time: 16 minutes


    Steroidogenic Factor 1/SF-1 contains a potential phosphorylation site, as well as a number of acetylation sites(SwissProt), which may explain its migration at a higher molecular weight than predicted. The 55 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to this target.

  • COS-1 whole cell lysate added at 20µg.
    Primary antibody used at a 1/1000 dilution.
    Secondary antibody used was a goat anti-rabbit IgG/ HRP at a 1/2500 dilution.

    See Abreview

  • IHC-P image of Steroidogenic Factor 1/SF-1 (ab65815) on Human testis sections. The sections were fixed in Paraformaldehyde and underwent heat mediated antigen retrieval using Novacastra (pH6). The sections were then blocked in 0.3% H2O2 solution for 10 mins at 22°C.

    See Abreview

  • All lanes : Anti-Steroidogenic Factor 1/SF-1 antibody (ab65815) at 1/900 dilution

    Lane 1 : Canine adrenocortical tumour whole cell lysate with ECL Blocking Solution as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4%
    Lane 2 : canine normal adrenal whole cell lysate with ECL Blocking Solution as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4%
    Lane 3 : H295R whole cell lysate with ECL Blocking Solution as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4%

    Secondary
    All lanes : Goat anti-Rabbit IgG, at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 52 kDa

    See Abreview

  • Formalin-fixed, paraffin-embedded dog adrenal cortex tissue stained for Steroidogenic Factor 1/SF-1 using ab65815 at 1/300 dilution in immunohistochemical analysis.

    See Abreview

References

This product has been referenced in:

  • Wu PL  et al. Transcription factor 21 regulates expression of ERß and SF-1 via upstream stimulatory factor-2 in endometriotic tissues. Biochim Biophys Acta Gene Regul Mech 1861:706-717 (2018). Read more (PubMed: 30018006) »
  • Lu Y  et al. Epigenetic modifications promote the expression of the orphan nuclear receptor NR0B1 in human lung adenocarcinoma cells. Oncotarget 7:43162-43176 (2016). Read more (PubMed: 27281610) »
See all 4 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Western blot
Sample
Dog Cell lysate - whole cell (Canine adrenocortical tumor, canine normal adrenal)
Gel Running Conditions
Non-reduced Denaturing (10% Criterion TGX Stain-Free)
Loading amount
20 µg
Specification
Canine adrenocortical tumor, canine normal adrenal
Blocking step
ECL Blockin Solution as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 21°C

Karin Sanders

Verified customer

Submitted Jan 23 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Adrenal cortex)
Antigen retrieval step
None
Permeabilization
No
Specification
Adrenal cortex
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Formaldehyde

Karin Sanders

Verified customer

Submitted Jan 23 2018

Answer

Thank you for confirming this information and for your help and cooperation with this case.

As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order.

Credit ID: 22303

As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

We would also appreciate if you are able to remind the team at Zotal that it is most helpful for us to have the order number within the first email when we are contacted regarding a complaint. This helps us to check the order is within guarantee period, that there were no problems with delivery, and is also crucial for our quality monitoring. We appreciate your consideration.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.

Read More

Question

Here are customer's answers:

1. I fully understand your concerns and it is highly unusual for 4 antibodies not to work for a customer. In this case, I can suggest to consider reviewing the sample preparation in particular. For example, try 1 fixing in 10% PFA for at least 24 hours.

The sample preparation is fine, and I have successfully stained these samples with multiple antibodies, some from abcam. These are rare samples of human tissue, and I cannot play around with the fixation.

2. Antigen retrieval can often require individual optimization for different antibodies with different epitopes. For example, I can recommend to try different time points for antigen retrieval. Try 2, 3, 5, 10 and 20 minutes.

I agree that antigen retrieval may be an issue. However, I did try two very different buffers, and two time points. Once again, I have a limited supply of samples, and cannot test all of these different time points.

3. As the target protein is nuclear, I can recommend to include a separate permeabilization step before staining.

I stained plenty of nuclear antigens on these same slides under the same conditions

4. I would suggest to try a lower concentration of antibody, 1:200 or even 1:500, to try and reduce the background.

Did that (1:500) - no improvement.

5. I can recommend to include 0.2% Tween in wash buffer to help keep the cells permeabilized and to wash away any non specific binding. Secondary antibody working well with other primary antibodies?

Secondary antibody works fine with other primaries. I don't normally have issues with non-specific binding.

6. 0.3% Triton is quite high, and may affect the staining. Try 0.2%.

As I wrote, I tried reducing and even eliminating triton, to no avail.

I have expended all of the sample slides available to me for finding the right conditions, and still have no results. This has cost great efforts and more resources than it should have. I will need to purchase a different antibody for this antigen from another company, so for now I guess I would like a credit note for a different antibody.

Thanks in advance foryour reply.

Best Regards,

Read More
Answer

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments, and would be pleased to provide the credit note in compensation as requested, providing the product has been purchased within the 6 month guarantee period. In order to arrange this, I would appreciate if you could confirm your order number and date of purchase?

Thank you for your continued cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Question

Dear Tech Support Team,

Please see below the details of our customer's complaint.

It is important to emphasize that this ab was supplied as the replacement for ab65815 that also did not perform well (the correspondence is attached as well).

Antibody code: ab103679

Batch number: GR29484-5

General Information:

Antibody storage conditions Stored at 4C for short period (aliquots in -20C)

Description of the problem No specific staining

Sample Parrafin sections of human and mouse testis, and rat ovaries

Fixation of sample Paraformaldehyde 4% for 24 hours

Antigen retrieval Heat induced, tried both citrate pH 6.0 and TE pH 9.0

Permeabilization step

Blocking conditions Blocked in buffer containing 0.3% Triton, 0.05% Tween-20 and 4% Normal Donkey Serum. Also tried same buffer without Triton. Blocked for 1 hour.

Primary Antibody Diluted antibody 1:50 or 1:100 in blocking buffer, incubated overnight at 4C. Washed with PBS.

Secondary Antibody Biotin conjugated antibody from Jackson immunoresearch. Diluted 1:200 in blocking buffer.

Detection method Cy2 or Cy3 conjugated avidin

Positive and negative controls used Other antibodies stained the same sections with no problem. No negative control as the issue is lack of signal

Optimization attempts (problem solving):

How many times have you tried the IHC? 6

Have you run a "No Primary" control? Yes – no issues.

Do you obtain the same results every time? Yes

What steps have you altered? I have altered the antigen retrieval buffers, and tried removing triton from the blocking buffer. Also tried different dilutions of the antibody

Additional Notes

SF1 is expressed in the steroidogenic cells of the testis (Sertoli) and ovaries (Granulosa). These cells are present in the tissue, and stain positive for other markers (ie Sox9 with abcam ab76997), but there is no staining whatsoever.

Document Attachment: images of the results may be very helpful. If you wish to add it to the complaint form, please attach them to the reply e-mail.. ATTACHED

Please read below customer's additional comments:

Does abcam have any other primary antibody against SF1 good for IHC in human?

This is the third out of 4 antibodies I have tried for this project that is failing. I am beginning to worry that maybe abcams QC isn't what it used to be.

The slides and reagents I use are obviously expensive, and it is frustrating to have to waste so many in an attempt to get any kind of staining. I always add a positive control to my experiments, so I know that both the slides and the assay work.

Kindly advise.

Thank you for your assistance and reply.

Kind Regards,

Read More
Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody and that you have had difficulties with previous products aswell.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab103679 is tested and covered by our 6 month guarantee for use in IHC-P and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results. I would also appreciate if you can confirm some further details:

1. I fully understand your concerns and it is highly unusual for 4 antibodies not to work for a customer. In this case, I can suggest to consider reviewing the sample preparation in particular. For example, try 1 fixing in 10% PFA for at least 24 hours.

2. Antigen retreival can often require individual optimization for different antibodies with different epitopes. For example, I can recommend to try different time points for antigen retrieval. Try 2, 3, 5, 10 and 20 minutes.

3. As the target protein is nuclear, I can recommend to include a separate permeabilization step before staining.

4. I would suggest to try a lower concentration of antibody, 1:200 or even 1:500, to try and reduce the background.

5. I can recommend to include 0.2% Tween in wash buffer to help keep the cells permeabilized and to wash away any non specific binding. Secondary antibody working well with other primary antibodies?

6. 0.3% Triton is quite high, and may affect the staining. Try 0.2%.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (testis)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9.0
Permeabilization
No
Specification
testis
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 05 2012

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Testis)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Novacastra, pH 6.0
Permeabilization
No
Specification
Testis
Blocking step
H2O2 as blocking agent for 10 minute(s) · Concentration: 0.3% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 29 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (COS-1 cells)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
COS-1 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Mr. Chun Fui Lai

Verified customer

Submitted Feb 24 2011

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