Overview

  • Product name

  • Description

    Mouse monoclonal to ARID1B
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment: PPAKRHEGDM YNMQYSSQQQ EMYNQYGGSY SGPDRRPIQG QYPYPYSRER MQGPGQIQTH GIPPQMMGGP LQSSSSEGPQ QNMWAARNDM PYPYQNR, corresponding to amino acids 1364-1461 of Human ARID1B

  • Positive control

    • ICC/IF: HepG2 cells.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 12 Feb 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab57461 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 at an assay dependent concentration. Predicted molecular weight: 236 kDa.
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Target

  • Relevance

    ARID1B (AT-rich interactive domain-containing protein 1B) is a component of SWI/SNF chromatin remodeling complexes involved in transcriptional activation and repression of select genes by chromatin remodeling. It binds DNA non-specifically and alters DNA-nucleosome topology.
  • Cellular localization

    Nuclear
  • Database links

  • Alternative names

    • 6A3 5 antibody
    • ARID 1B antibody
    • ARID domain containing protein 1B antibody
    • AT rich interactive domain 1B (SWI1 like) antibody
    • AT rich interactive domain 1B antibody
    • AT rich interactive domain containing protein 1B antibody
    • BAF 250b antibody
    • BAF-associated factor, 250-KD, B antibody
    • BAF250b antibody
    • BRG1 associated factor 250b antibody
    • BRG1 binding protein antibody
    • BRG1 binding protein ELD/OSA1 antibody
    • BRG1 binding protein hELD/OSA1 antibody
    • BRIGHT antibody
    • DAN 15 antibody
    • DAN15 antibody
    • Eld (eyelid)/Osa protein antibody
    • ELD/OSA1 antibody
    • hELD/OSA1 antibody
    • hOsa 2 antibody
    • hOsa2 antibody
    • KIAA1235 antibody
    • MRD12 antibody
    • OSA 2 antibody
    • Osa homolog 2 antibody
    • OSA2 antibody
    • OTTHUMP00000040115 antibody
    • p250R antibody
    • RP11 419L10.1 antibody
    see all

Images

  • ICC/IF image of ab57461 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab57461, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    This image was generated using the ascites version of the product.

  • ARID1B antibody (ab57461) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human kidney.

    This image was generated using the ascites version of the product.

  • ARID1B antibody (ab57461) at 1ug/lane + HeLa cell lysate at 25ug/lane.

    This image was generated using the ascites version of the product.

  • Overlay histogram showing HEK293 cells stained with ab57461 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab57461, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    This image was generated using the ascites version of the product.

References

This product has been referenced in:

  • Cui Y  et al. Upregulated expression of AT-rich interactive domain-containing protein 1B predicts poor prognosis in patients with triple-negative breast cancer. Oncol Lett 17:3289-3295 (2019). Read more (PubMed: 30867762) »
  • Raab JR  et al. SWI/SNF remains localized to chromatin in the presence of SCHLAP1. Nat Genet 51:26-29 (2019). Read more (PubMed: 30510238) »
See all 13 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for contacting us.

Your credit note ID is 23074.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

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

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA pH8.5
Permeabilization
No
Specification
kidney
Blocking step
Ventana blocking agent as blocking agent for 4 minute(s) · Concentration: 100% · Temperature: 36°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 07 2015

Question

Lot number: GR94192-1
Inquiry: GENERAL INFORMATION Antibody Storage Conditions (temperature/reconstitution etc) As instructed. -20°C and freeze/thaw was avoided Description of the Problem (high background, wrong band size, more bands, no bands etc) Wrong band size. 2 non-specific bands between 52kDa and 30kDa Sample (Species/Cell extract/Purified protein/Recombinant protein, etc) HEK293FT, BE2-M17 neuroblastoma and human fibroblasts lysates. Sample Preparation (Buffer/Protease inhibitors/Heating sample etc.) Lysed immediately with 50mM Tris, pH 6.8, 5% SDS, 50mM EDTA on the culture plate. Sample was then sonicated to shear genomic DNA. Protein concentration was measured and appropriate amount was then mixed with the required volume of 5X SDS-PAGE sample loading buffer Amount of protein loaded 80ug Electrophoresis/Gel Conditions (Reducing or non-reducing gel, % of the gel etc.) Reducing, 10% polyacrylamide Transfer and blocking conditions (Buffer/time period, Blocking agent etc) 10v, overnight wet transfer for 16.5 hrs. Blocking agent, 5% BSA in TBST, for 30min Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step) Abcam/ Host species: mouse target species: human/ diluents: 1% BSA in TBST/ Dilution 1:500/ incubation time: overnight at 4°C/ Wash: 3x 5min wash with TBST Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step) Manufacturer: Dako/ Species: rabbit/ Diluent: 1% BSA in TBST/ Dilution: 1:8000/ Incubation time: 1hr/ Wash: 3x 5min wash with TBST Detection Method (ECL, ECL Plus etc.) ECL Plus Positive and negative controls used (please specify) No available positive control Plausible negative control: Patient fibroblast with only one copy of ARID1B OPTIMIZATION ATTEMPTS (PROBLEM SOLVING) How many times have you tried the Western Blot? 5 Have you run a “No Primary” control? Yes Do you obtain the same results every time? Yes e.g. are the background bands always in the same place? What steps have you altered? Extending incubation time, and trying other cell lysates Comments from Sapphire: Sapphire Order #:PO-15546 Abcam Invoice #: 1350925B Not sure of exact lot # but this is the first time we have ordered this product.

Read More
Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

- Could you confirm if the immunoprecipitation has been done or not?
- Have you used any loading control?

Suggestions for further improvements;

-Could you try the lysates without sonicating?

Thank you very much for your cooperation. I will look forward to hearing from you soon.

Read More
Application
Western blot
Sample
Human Cell lysate - nuclear (JHH4)
Loading amount
10 µg
Specification
JHH4
Treatment
Use nuclear fraction or cytoplasmic fraction
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 20 2011

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