Overview

  • Product name
  • Description
    Rabbit polyclonal to SNF5
  • Host species
    Rabbit
  • Specificity
    This antibody is specific for SMARCB1.
  • Tested applications
    Suitable for: ICC/IF, ELISA, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Horse, Guinea pig, Cow, Dog
  • Immunogen

    A region within synthetic peptide: RGSLYKRYPS LWRRLATVEE RKKIVASSHG KKTKPNTKDH GYTTLATSVT, corresponding to amino acids 40-89 of Human SNF5

  • Positive control
    • Human liver tissue and fetal thymus lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab42503 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 1/150.
ELISA 1/312500.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use a concentration of 4 - 8 µg/ml.

Target

  • Function
    Core component of the BAF (hSWI/SNF) complex. This ATP-dependent chromatin-remodeling complex plays important roles in cell proliferation and differentiation, in cellular antiviral activities and inhibition of tumor formation. The BAF complex is able to create a stable, altered form of chromatin that constrains fewer negative supercoils than normal. This change in supercoiling would be due to the conversion of up to one-half of the nucleosomes on polynucleosomal arrays into asymmetric structures, termed altosomes, each composed of 2 histones octamers. Stimulates in vitro the remodeling activity of SMARCA4/BRG1/BAF190A. Involved in activation of CSF1 promoter. Belongs to the neural progenitors-specific chromatin remodeling complex (npBAF complex) and the neuron-specific chromatin remodeling complex (nBAF complex). During neural development a switch from a stem/progenitor to a post-mitotic chromatin remodeling mechanism occurs as neurons exit the cell cycle and become committed to their adult state. The transition from proliferating neural stem/progenitor cells to post-mitotic neurons requires a switch in subunit composition of the npBAF and nBAF complexes. As neural progenitors exit mitosis and differentiate into neurons, npBAF complexes which contain ACTL6A/BAF53A and PHF10/BAF45A, are exchanged for homologous alternative ACTL6B/BAF53B and DPF1/BAF45B or DPF3/BAF45C subunits in neuron-specific complexes (nBAF). The npBAF complex is essential for the self-renewal/proliferative capacity of the multipotent neural stem cells. The nBAF complex along with CREST plays a role regulating the activity of genes essential for dendrite growth (By similarity). Plays a key role in cell-cycle control and causes cell cycle arrest in G0/G1. Also involved in vitamin D-coupled transcription regulation via its association with the WINAC complex, a chromatin-remodeling complex recruited by vitamin D receptor (VDR), which is required for the ligand-bound VDR-mediated transrepression of the CYP27B1 gene.
  • Involvement in disease
    Defects in SMARCB1 are a cause of rhabdoid tumor (RDT) [MIM:609322]; also known as malignant rhabdoid tumor (MRT). RDT are a highly malignant group of neoplasms that usually occur in early childhood. SMARCB1/INI1 is also frequently inactivated in epithelioid sarcomas.
    Defects in SMARCB1 are a cause of schwannomatosis (SCHWA) [MIM:162091]; also called congenital cutaneous neurilemmomatosis. Schwannomas are benign tumors of the peripheral nerve sheath that usually occur singly in otherwise normal individuals. Multiple schwannomas in the same individual suggest an underlying tumor-predisposition syndrome. The most common such syndrome is NF2. The hallmark of NF2 is the development of bilateral vestibular-nerve schwannomas; but two-thirds or more of all NF2-affected individuals develop schwannomas in other locations, and dermal schwannomas may precede vestibular tumors in NF2-affected children. There have been several reports of individuals with multiple schwannomas who do not show evidence of vestibular schwannoma. Clinical report suggests that schwannomatosis is a clinical entity distinct from other forms of neurofibromatosis.
  • Sequence similarities
    Belongs to the SNF5 family.
  • Post-translational
    modifications
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • BAF47 antibody
    • BRG1-associated factor 47 antibody
    • hSNF5 antibody
    • INI1 antibody
    • Integrase interactor 1 protein antibody
    • Malignant rhabdoid tumor suppressor antibody
    • RDT antibody
    • RTPS1 antibody
    • Sfh1p antibody
    • SMARCB1 antibody
    • SNF5 homolog antibody
    • SNF5_HUMAN antibody
    • SNF5L1 antibody
    • Snr1 antibody
    • Sucrose nonfermenting yeast homolog like 1 antibody
    • SWI/SNF complex component SNF5 antibody
    • SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily B member 1 antibody
    • SWI10 antibody
    • Transcription factor TYE4 antibody
    • Transcription regulatory protein SNF5 antibody
    • TYE4 antibody
    see all

Images

  • Anti-SNF5 antibody (ab42503) at 2 µg/ml + fetal thymus lysate

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 44 kDa
    Observed band size: 44 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling SNF5 with ab42503 at 4-8µg/ml.

References

ab42503 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. I have checked with my lab colleagues who confirmed the epitopes were retrieved by incubating the sections at 98-100C in a microwave in 0.01 M citrate buffer (pH 6.0) for 15-20 minutes. Primary antibody incubation was performed at RT for 45minutes.

You can also find very useful information and protocol tips in our online protocols:

https://www.abcam.com/index.html?pageconfig=resource&rid=11384

In any case, if after repeating the experiment following the protocol described the antibody does not yet give the expected signal, I would encourage you to contact us back and we will happily offer you more assistance.

All of our products are covered by our Abpromise® guarantee, which ensures that you can trust our products, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 6 months of purchase.

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Answer

Thank you for contacting us and sorry for the delay in getting back to you. The following protocol was used to perform the staining presented on the datasheet of the anti-SNF5 antibody (ab42503):

Tissue fixation and preparation

1. Tissue fixed with formalin then embed in paraffin wax to dehydrate.

2. The tissue was cutin 4 µm sections on charged slides and heated in oven for 45 min at 60°C

3. The slides were then washed 3X in xylene for 5 min at RT

4. Slides were then rehydrated in 2X in 95% alcohol for 3 min at RT, 1X in 80% alcohol for 3 min at RT. The slides were then gently rinsed using distilled H2O for 5 min at RT andendogenous peroxidase quenched for 10 min with 3% H2O2 in 0.1 M PBS and wash 3 times with distilled water, 2 minutes each.

Antigen retrieval:

1.The epitope was retrieved by incubating the sections at 98-100° C in a microwave in 0.01 M citrate buffer (pH 6.0) for 15-20 minutes. The slides were then allowed to cool down atroom temperature for 30-60 min.

2. Slides rinsed with1X in 0.1% TBST for 1 min at RT

Immunostaining:

1.Protein block (10% normal goat serum) was applied for 20 min at RT thendrained.

2. Primary antibody was applied (1/125 diluted in 10% normal goatserum)for 45 min at RT then rinsed 3X for 1 min in 0.1% TBST.

3.Diluted biotinylated secondary antibody was then applied for 45 min at RT, thenrinsed 3X for 1 min in 0.1% TBST.

4. AP-streptavidin applied for 30 min at RT,rinsed 3X for 1 min in 0.1% TBST.

5.Alkaline chromogen substrate applied for 30 min at RT, rinsed 1X in dH2O

6.Counterstain added for 10-30 seconds with a hematoxylin counterstaining solution, rinsed 1X in dH2O

Dehydration

1. Slides washed in 2X in 80% alcohol for 1 min, 2X in 95% alcohol for 1 min, 3X in xylene for 1 min all atRT.

2.Coverslip added with mounting medium

I hope this information has been of help. If you would like for me to have a look at the procedure you have been using and see if there are any suggestions I can make, please could you fill out the questionnaire I have attached to this email.

If the product was purchased in the last six months and is being used according to our Abpromise, and you are not observing the results as expected, we would be happy to replace or refund the antibody.


I look forward to receiving your reply.

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Question

This email has 2 sections:

A recap of the complaint

The respondent’s survey results (section-by-section)
Recap of the complaint
---------------------------------------------------------------------------------------------------------------------


Date(s) submitted: February 22, 2012

Catalog number(s): 42503

Product(s): Rabbit polyclonal to SNF5

CCEID: 3532783
Survey results
---------------------------------------------------------------------------------------------------------------------

Telephone section
Q. Did you contact Abcam's Scientific Support by telephone in reference to this complaint?
A. No
Q. Was it clear from the phone options which number you needed to press to get your issue resolved?
A.

Protocol advice section
Q. Did Abcam's Scientific Support give you protocol advice?
A.Yes
Q. How did you feel about being given protocol advice?
A.I requested protocol advice
Q. Did the product work successfully after following the advice?
A.No
Q. Why have you not tried the protocol advice? (Select all that apply.)
A.
A.
A.
A.

Free of charge replacement section
Q. Did Abcam's Scientific Support give you a free of charge replacement?
A. Yes

Q. Did the replacement product work successfully?
A.No
Credit note / refund section
Q. Did Abcam's Scientific Support give you a credit note or a refund?
A.Yes

Q. Would you have preferred a different outcome from receiving a credit note or refund?
A.buy an antibody that works
Overall satisfaction section
Q. How satisfied are you with how the complaint was handled?
A.Neither satisfied nor unsatisfied
Q. Could we have done anything to better resolve the problem?
A.Yes, sell antibody actually working. It's too easy to sell any kind of "shitty" product and then give a refund to the customer...
Q. Would you mind if our scientific support team contacted you to better understand what happened and reach an agreeable solution?
A.No, I would not mind
Q. Please provide a phone number or email address where we can reach you.
A.
Q. Please share with us any final thoughts you may have about Abcam, the complaints process, or our products.
A.The last 3 antibodies i bought, none of them worked. It's too easy to sell any kind of "shitty" product and then give a refund to the customer

Best Regards,

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Answer

Thank you for sharing your views and for submitting your survey's results. It is very unfortunate that this antibody (ab42503:Anti-SNF5 antibody) does not perform as it is expected.
All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.
We have been selling this antibody since 2007 and according to our record; this is the first complaint so we are not aware of any quality issues.
Once again, thank you for your valuable feedback!
If you need any further assistance in the future, please do not hesitate to contact me.

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Answer

Thank you for contacting us.
Your credit note ID is 19856.
I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.
Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.
The credit note ID is for your reference only and does not automatically guarantee the credit.
I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Answer

Thank you for getting back to me promptly and for answering to my further questions and for attaching the images. Your time and effort is much appreciated!
It is clear that ab42503 does not detect SNF5, it may well be that it has been damaged accidenatlyy either during shipment or storage.
The protocol used to test this antibody is a standard IHC procedure (briefly).
- Retrieve epitopes by incubating the sections at 98-100° C in a microwave in 0.01 M citrate buffer (pH 6.0) for 15-20 minutes. Cool down in room temperature for 30-60 min.
- Apply universal protein block for 20 min at RT; drain
- The primary antibody concentration for our IHC image on human liver tissue was a range; between 4-8 ug/mL.
- Detection: ABC based signal enhancing step - biotinylated secondary antibody for 45 min at RT; rinse 3X for 1 min in 0.1% TBST, Apply AP-streptavidin for 30 min at RT; rinse 3X for 1 min in 0.1% TBST
I could certainly offer you either a credit note which you can use in future purchase or a free of charge replacement. Please do let me know how you wish to proceed with your enquiry.
I look forward to hearing from you soon.

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Answer

Thank you for your enquiry regarding ab42503 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.
As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.
Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem.
1) Image:
- Could you attach an image (saved as a jpeg file) to your e-mail so that I can have a look at the signal.
2) Fixation:
- It would help me if you could clarify how long the samples were fixed?
- Has perfusion fixation been carried out?
- What percentage of Tween was used for permeabilization?
3) Antigen retrieval:
- How long the antigen retrieval was performed?
- Which technique was used (microwave, pressure cooker or steamer)?
4) Order Number:
- It would be much appreciated if you could confirm the Abcam order number (or your PON) and the date of purchase.
Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cultured Cells (3T3)
Specification
3T3
Fixative
Formaldehyde
Permeabilization
Yes - 0.5% triton X100
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5%

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Submitted Apr 22 2010

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