Product Info Summary
SKU: | A00223-1 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-BRG1/SMARCA4 Antibody Picoband®
SKU/Catalog Number
A00223-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-BRG1/SMARCA4 Antibody Picoband® catalog # A00223-1. Tested in ELISA, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-BRG1/SMARCA4 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00223-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human BRG1 recombinant protein (Position: Q555-E763).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00223-1 is reactive to SMARCA4 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
220 kDa
Calculated molecular weight
184.646kDa
Background of Brg1
Transcription activator BRG1 also known as ATP-dependent helicase SMARCA4 is a protein that in humans is encoded by the SMARCA4 gene. The protein encoded by this gene is a member of the SWI/SNF family of proteins and is similar to the brahma protein of Drosophila. Members of this family have helicase and ATPase activities and are thought to regulate transcription of certain genes by altering the chromatin structure around those genes. The encoded protein is part of the large ATP-dependent chromatin remodeling complex SNF/SWI, which is required for transcriptional activation of genes normally repressed by chromatin. In addition, this protein can bind BRCA1, as well as regulate the expression of the tumorigenic protein CD44. Mutations in this gene cause rhabdoid tumor predisposition syndrome type 2. Multiple transcript variants encoding different isoforms have been found for this gene.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00223-1 is guaranteed for ELISA, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Immunocytochemistry/Immunofluorescence, 2μg/ml
ELISA, 0.1-0.5μg/ml
Positive Control
WB: human Hela whole cell, human placenta tissue, human MCF-7 whole cell, human A549 whole cell, human A431 whole cell, human SGC-7901 whole cell, human 22RV1 whole cell, rat small intestine tissue, mouse small intestine tissue
IHC: human intestinal cancer tissue, mouse small intestine tissue, rat small intestine tissue
ICC/IF: U20S cell, U20S cell
FCM: U20S cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of BRG1 using anti-BRG1 antibody (A00223-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human placenta tissue lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: human A431 whole cell lysates,
Lane 6: human SGC-7901 whole cell lysates,
Lane 7: human 22RV1 whole cell lysates,
Lane 8: rat small intestine tissue lysates,
Lane 9: mouse small intestine tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-BRG1 antigen affinity purified polyclonal antibody (Catalog # A00223-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for BRG1 at approximately 220KD. The expected band size for BRG1 is at 185KD.
Click image to see more details
Figure 2. IHC analysis of BRG1 using anti-BRG1 antibody (A00223-1).
BRG1 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-BRG1 Antibody (A00223-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of BRG1 using anti-BRG1 antibody (A00223-1).
BRG1 was detected in paraffin-embedded section of mouse small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-BRG1 Antibody (A00223-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of BRG1 using anti-BRG1 antibody (A00223-1).
BRG1 was detected in paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-BRG1 Antibody (A00223-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. IF analysis of BRG1 using anti-BRG1 antibody (A00223-1)
BRG1 was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-BRG1 Antibody (A00223-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 6. IF analysis of BRG1 using anti-BRG1 antibody (A00223-1)
BRG1 was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-BRG1 Antibody (A00223-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 7. Flow Cytometry analysis of U20S cells using anti-BRG1 antibody (A00223-1).
Overlay histogram showing U20S cells stained with A00223-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-BRG1 Antibody (A00223-1,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For SMARCA4 (Source: Uniprot.org, NCBI)
Gene Name
SMARCA4
Full Name
Transcription activator BRG1
Weight
184.646kDa
Superfamily
SNF2/RAD54 helicase family
Alternative Names
Transcription activator BRG1; TP-dependent helicase SMARCA4; BRG1-associated factor 190A; BAF190A; Mitotic growth and transcription activator; Protein BRG-1; Protein brahma homolog 1; SNF2-beta; SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 4; SMARCA4; BAF190A; BRG1; SNF2B; SNF2L4 SMARCA4 BAF190, BAF190A, BRG1, CSS4, MRD16, RTPS2, SNF2, SNF2-beta, SNF2L4, SNF2LB, SWI2, hSNF2b SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4 transcription activator BRG1|ATP-dependent helicase SMARCA4|BRG1-associated factor 190A|BRM/SWI2-related gene 1|SNF2-like 4|brahma protein-like 1|global transcription activator homologous sequence|homeotic gene regulator|mitotic growth and transcription activator|nuclear protein GRB1|protein BRG-1|protein brahma homolog 1|sucrose nonfermenting-like 4
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on SMARCA4, check out the SMARCA4 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for SMARCA4: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-BRG1/SMARCA4 Antibody Picoband® (A00223-1)
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Customer Q&As
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16 Customer Q&As for Anti-BRG1/SMARCA4 Antibody Picoband®
Question
Our lab used your anti-BRG1/SMARCA4 antibody for IHC-P on lung in the past. I am using rat, and We intend to use the antibody for ICC next. I am interested in examining lung as well as embryonic kidney in our next experiment. Do you have any suggestion on which antibody would work the best for ICC?
Verified Customer
Verified customer
Asked: 2020-02-25
Answer
I have checked the website and datasheets of our anti-BRG1/SMARCA4 antibody and it seems that A00223-1 has been tested on rat in both IHC-P and ICC. Thus A00223-1 should work for your application. Our Boster satisfaction guarantee will cover this product for ICC in rat even if the specific tissue type has not been validated. We do have a comprehensive range of products for ICC detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2020-02-25
Question
I am looking for using your anti-BRG1/SMARCA4 antibody for positive regulation of transcription studies. Has this antibody been tested with western blotting on 22rv1 whole cell lysates? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2019-12-18
Answer
I appreciate your inquiry. This A00223-1 anti-BRG1/SMARCA4 antibody is tested on human placenta tissue, hela whole cell lysates, a549 whole cell lysates, a431 whole cell lysates, 22rv1 whole cell lysates, rat small intestine tissue lysates, intestinal cancer tissue, mouse small intestine tissue lysates, u20s cells. It is guaranteed to work for ELISA, IF, IHC-P, ICC, WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2019-12-18
Question
We have seen staining in rat fetal brain. What should we do? Is anti-BRG1/SMARCA4 antibody supposed to stain fetal brain positively?
Verified Customer
Verified customer
Asked: 2019-09-20
Answer
According to literature fetal brain does express SMARCA4. According to Uniprot.org, SMARCA4 is expressed in tendon of biceps brachii, fetal brain, lung, cervix carcinoma, embryonic kidney, leukemic t-cell, cervix carcinoma erythroleukemia, liver, among other tissues. Regarding which tissues have SMARCA4 expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, Pubmed ID: 16964243, 17081983, 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Embryonic kidney, Pubmed ID: 17525332
Fetal brain, Pubmed ID: 8208605
Leukemic T-cell, Pubmed ID: 19690332
Liver, Pubmed ID: 24275569
Lung, Pubmed ID: 18386774
Boster Scientific Support
Answered: 2019-09-20
Question
Do you have a BSA free version of anti-BRG1/SMARCA4 antibody A00223-1 available?
Verified Customer
Verified customer
Asked: 2019-09-16
Answer
I appreciate your recent telephone inquiry. I can confirm that some lots of this anti-BRG1/SMARCA4 antibody A00223-1 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2019-09-16
Question
I would like to test anti-BRG1/SMARCA4 antibody A00223-1 on rat cervix carcinoma for research purposes, then I may be interested in using anti-BRG1/SMARCA4 antibody A00223-1 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2019-08-13
Answer
The products we sell, including anti-BRG1/SMARCA4 antibody A00223-1, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2019-08-13
Question
Will anti-BRG1/SMARCA4 antibody A00223-1 work for IF with cervix carcinoma?
Verified Customer
Verified customer
Asked: 2019-06-20
Answer
According to the expression profile of cervix carcinoma, SMARCA4 is highly expressed in cervix carcinoma. So, it is likely that anti-BRG1/SMARCA4 antibody A00223-1 will work for IF with cervix carcinoma.
Boster Scientific Support
Answered: 2019-06-20
Question
We are currently using anti-BRG1/SMARCA4 antibody A00223-1 for human tissue, and we are happy with the IF results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on primate tissues as well?
P. Krishna
Verified customer
Asked: 2019-05-10
Answer
The anti-BRG1/SMARCA4 antibody (A00223-1) has not been validated for cross reactivity specifically with primate tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in primate you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-05-10
Question
Will A00223-1 anti-BRG1/SMARCA4 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2019-02-21
Answer
You can see on the product datasheet, A00223-1 anti-BRG1/SMARCA4 antibody as been tested on IF. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2019-02-21
Question
My boss were happy with the WB result of your anti-BRG1/SMARCA4 antibody. However we have been able to see positive staining in cervix carcinoma erythroleukemia nucleus using this antibody. Is that expected? Could you tell me where is SMARCA4 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2018-08-21
Answer
From literature, cervix carcinoma erythroleukemia does express SMARCA4. Generally SMARCA4 expresses in nucleus. Regarding which tissues have SMARCA4 expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, Pubmed ID: 16964243, 17081983, 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Embryonic kidney, Pubmed ID: 17525332
Fetal brain, Pubmed ID: 8208605
Leukemic T-cell, Pubmed ID: 19690332
Liver, Pubmed ID: 24275569
Lung, Pubmed ID: 18386774
Boster Scientific Support
Answered: 2018-08-21
Question
Is this A00223-1 anti-BRG1/SMARCA4 antibody reactive to the isotypes of SMARCA4?
Verified Customer
Verified customer
Asked: 2018-08-21
Answer
The immunogen of A00223-1 anti-BRG1/SMARCA4 antibody is E. coli-derived human BRG1 recombinant protein (Position: Q555-E763). Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2018-08-21
Question
Is a blocking peptide available for product anti-BRG1/SMARCA4 antibody (A00223-1)?
Verified Customer
Verified customer
Asked: 2018-08-21
Answer
We do provide the blocking peptide for product anti-BRG1/SMARCA4 antibody (A00223-1). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2018-08-21
Question
See attached the WB image, lot number and protocol we used for cervix carcinoma using anti-BRG1/SMARCA4 antibody A00223-1. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2018-06-13
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2018-06-13
Question
I see that the anti-BRG1/SMARCA4 antibody A00223-1 works with IF, what is the protocol used to produce the result images on the product page?
J. Carter
Verified customer
Asked: 2017-02-21
Answer
You can find protocols for IF on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2017-02-21
Question
I was wanting to use your anti-BRG1/SMARCA4 antibody for IF for rat cervix carcinoma on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for rat cervix carcinoma identification?
L. Roberts
Verified customer
Asked: 2016-06-20
Answer
It shows on the product datasheet, A00223-1 anti-BRG1/SMARCA4 antibody has been validated for ELISA, IF, IHC-P, ICC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in rat cervix carcinoma in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2016-06-20
Question
I appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for cervix carcinoma using anti-BRG1/SMARCA4 antibody A00223-1. Let me know if you need anything else.
R. Miller
Verified customer
Asked: 2016-03-09
Answer
Thanks for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2016-03-09
Question
My question regarding product A00223-1, anti-BRG1/SMARCA4 antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
P. Bhatt
Verified customer
Asked: 2013-02-28
Answer
We suggest not storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free A00223-1 anti-BRG1/SMARCA4 antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2013-02-28