BLIMP1/PRDM1 Antibody (3H2-E8) 0.1 ml Plasma Cell Marker

Click image to view larger
Western Blot: Blimp-1 Antibody (3H2-E8) [NB600-235] - Blimp-1 expression by CD19+ (thick black line) or CD8+ (thin, shaded line). (Tedder Lab; Duke Univ)
Immunocytochemistry/Immunofluorescence: Blimp-1 Antibody (3H2-E8) [NB600-235] - Double IF for Blimp-1 (green) and Ki-67 (proliferation, red) with DAPI counterstain, at 10x magnification (scale bar is 10um).  Positive surface epithelium and centrocytes are labelled.   Photo courtesy of Dr. Giorgio Cattoretti, Institute for Cancer Genetics, Columbia University, NY.
Flow Cytometry: Blimp-1 Antibody (3H2-E8) [NB600-235] - Blimp-1 expression by IL-10+ (black line), IL-10- (dotted line) or CD8+ (thin, shaded line) cells (Tedder Lab; Duke Univ)
Western Blot: Blimp-1 Antibody (3H2-E8) [NB600-235] - Detection of Blimp-1 in murine plasmacytoma cell lysate (P3X) using NB 600-235. 1881: murine pre-B cell lysate (negative control). Photo courtesy of DA Savitsky, Columbia University.
Flow Cytometry: Blimp-1 Antibody (3H2-E8) [NB600-235] - IL-10+Blimp-1-, IL-10+Blimp-1+, IL-10-Blimp-1+ and IL-10-Blimp-1- cells, % total living single splenic CD19+ cells of 3 total mice (Tedder Lab; Duke Univ).
Download Datasheet
See All Related

Ordering Information: NB600-235

  • Catalog Number
    NB600-235
  • Displayed Format
    Unconjugated
  • Other Formats
  • Size(s)
  • Availability
  • Price
    Shipping
  

BLIMP1/PRDM1 Antibody (3H2-E8) Summary

Species Human, Mouse, Porcine
Tested Applications WB, ChIP, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP
Clonality
Monoclonal
Host
Mouse
Gene
PRDM1
Purity
Protein G purified
Guarantee Plus
All of our products are backed by our 100% guarantee to work for stated and predicted species and applications.
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

BLIMP1/PRDM1 Antibody (3H2-E8) Details

Clone
3H2-E8
Isotype
IgG1
Marker
Plasma Cell Marker
Immunogen
A fragment of mouse Blimp-1 corresponding to residues 199-409. [UniProt# Q60636]
Localization
Nuclear

Species Reactivity

Human, mouse and pig.
Reviewed Species Read 3 rated 3.3
using NB600-235 in the following species:

Publications Read Publications using NB600-235 in the following species:


Applications/Dilutions

Dilutions
  • Western Blot 1:1000
  • Chromatin Immunoprecipitation 1ug antibody / 10 ug protein
  • ELISA 1:100-1:2000
  • Flow Cytometry 8 ug/ml where cells are used at 2*10^6/mL
  • Immunocytochemistry/Immunofluorescence 1:50-1:200
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200
  • Immunoprecipitation
Application Notes
This Blimp-1 (3H2-E8) antibody is useful for ELISA, Immunocytochemistry/Immunofluorescence, Western Blot, Immunohistochemistry on paraffin-embedded sections, Flow Cytometry, Immunoprecipitation and Chromatin Immunoprecipitation applications. Immunohistochemistry-Frozen was reported in scientific literature. By WB, this antibody recognizes a band at ~98 kDa and may recognize one at ~80 kDa (the beta form). Antigen retrieval is recommended (EDTA buffer, microwave) prior to IHC on paraffin tissues. This antibody demonstrates nuclear staining. For IHC and ICC/IF a dilution of 1:200 is recommended with tyramide amplification.
Positive Controls
Other Available
Formats
7C Labeled NB600-235V2
Alexa Fluor (R) 405 Labeled NB600-235AF405
Alexa Fluor (R) 488 Labeled NB600-235AF488
Alexa Fluor (R) 647 Labeled NB600-235AF647
Alexa Fluor (R) 700 Labeled NB600-235AF700
Allophycocyanin Labeled NB600-235APC
Biotin Labeled NB600-235B
DyLight 350 Labeled NB600-235UV
DyLight 405 Labeled NB600-235V
DyLight 405LS Labeled NB600-235V3
DyLight 488 Labeled NB600-235G
DyLight 550 Labeled NB600-235R
DyLight 650 Labeled NB600-235C
DyLight 680 Labeled NB600-235FR
DyLight 755 Labeled NB600-235IR
FITC Labeled NB600-235F
HRP Labeled NB600-235H
PE Labeled NB600-235PE
PerCP Labeled NB600-235PCP
Reviewed Applications
Read 3 Review rated 3.3
using
NB600-235 in the following applications:

Publications
Read Publications using
NB600-235 in the following applications:

Contact Information

Product PDFs

Frequently Purchased Together

Reviews

Earn rewards points by submitting your review for BLIMP1/PRDM1 Antibody (NB600-235).

Publications

BLIMP1/PRDM1 Antibody (NB600-235) has been mentioned in at least 36 publications.
Earn rewards if you have published using BLIMP1/PRDM1 Antibody (NB600-235).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
Tris-glycine, 150 mM NaCl
Unit Size
0.1 ml
Concentration
1 mg/ml
Preservative
0.1% Sodium Azide
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

Gene Symbol PRDM1
Entrez
Uniprot

Alternate Names for BLIMP1/PRDM1 Antibody (3H2-E8)

  • Beta-interferon gene positive regulatory domain I-binding factor
  • beta-interferon gene positive-regulatory domain I binding factor
  • BLIMP-1
  • BLIMP1MGC118925
  • B-lymphocyte-induced maturation protein 1
  • MGC118922
  • MGC118923
  • Positive regulatory domain I-binding factor 1
  • PR domain containing 1, with ZNF domain
  • PR domain zinc finger protein 1
  • PR domain-containing protein 1
  • PRDI-BF1MGC118924
  • PRDI-binding factor 1
  • PRDI-binding factor-1
  • PR-domain zinc finger protein 1
Show More

Related Products by Gene

Background

Blimp-1 (B lymphocyte-induced maturation protein) is a nuclear zinc-finger containing transcriptional repressor which primarily controls the terminal differentiation of mature B cells to plasma cells, and also involves in macrophage as well as T-cells population homeostasis/differentiation. It is also called positive regulatory domain I-binding factor-1 (PRDI-BF1) or PR (PRDI-BF1-RIZ) domain zinc finger protein 1 (PRDM1) and its first human homolog, PRDI-BF1, was identified by its ability to bind to the PRDI element on the IFN-beta promoter leading to inhibition of virus-mediated IFN-beta production. Blimp-1 contains N-terminal PR/SET domain and five C2H2 zinc fingers located near its C-terminus that mediate DNA binding, nuclear import and recruitment of histone modifying enzymes, and these activities enable Blimp-1 for its ability to control cell-fate decisions in the embryo and govern tissue homeostasis in multiple cell types in the adult organism. Blimp-1 recruits chromatin-modifying enzymes including HDACs and methyltransferases, and some of its target genes include c-Myc, CIITA, Pax5, Spi-B, and Id3. Blimp-1 sumoylation at Lys-816 by PIAS1 augments transcriptional repressor activity, and is critical for plasma cell differentiation. Deletion mutation of Blimp-1 is frequently observed in several tumors including lymphoid malignancies and has been proposed to be a candidate of tumor suppressor gene.

Reviews for BLIMP1/PRDM1 Antibody (NB600-235) (3) 3.33

Read what people are saying who have used BLIMP1/PRDM1 Antibody (NB600-235).

We have 3 reviews tested in 1 species: Other.
We have 3 reviews tested in 3: Immunohistochemistry-Paraffin, Immunocytochemistry/Immunofluorescence, Immunofluorescence.
Average Rating: 3.3
(Based on 3 reviews)

Have you used BLIMP1/PRDM1 Antibody (NB600-235)? Submit your review and earn rewards points which can be used for merchandise & discounts.

Sort 3 reviews for BLIMP1/PRDM1 Antibody (NB600-235) by:
Filter by Ratings
Very Good
(1)
Average
(2)
All Ratings
Filter by Applications
IHC-P
(1)
ICC/IF
(2)
All Applications
Filter by Species
Other
(3)
All Species

Images Ratings Applications Species Reviewed By Date Details
  Very Good
IHC-P Other 05/31/2010
Show

Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedPorcine embryo
SpeciesOther
Other SpeciesPorcine
CommentsI stained paraffin embedded porcine embryonic tissue to verify the usefulness of Blimp1 as a germline marker. The staining was highly specific (nuclear), but only successful when using chromogenic development (used the HRP-based AEC-red system). Citrate buffer was superior to EDTA for antigen retrieval and 1:500 dilution in 0.05M Tris-HCl, pH 6.0 with 1% BSA was superior to higher concentrations and higher pH or PBS-based solution.  Detection Method: AEC red. Negative Control: no primary and isotype control. Deparaffinization: 2x10min xylene, 2x 5min 99% etOH, 3min 96% etOH, 3min 70% etOH, 3 min water. Antigen Retrieval: 0.01M citrate buffer, 15min at boiling.

Blocking

Blocking DetailsBlocking Buffer: 2% BSA, Blocking Time: 10 minutes, Blocking Temp: room temperature

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:500, Incubation Dilution Buffer: 0.05M Tris-HCL, pH 6.0, 1 % BSA, Incubation Time: 1hr, Temp: RT

Details

Detection NotesPlease see comments
Fixation DetailsPFA fixed and paraffin embedded, Deparaffinization: 2x 10min xylene, 2x 5min 99% etOH, 3min 96% etOH, 3min 70% etOH, 3min water

Comments

CommentsI stained paraffin embedded porcine embryonic tissue to verify the usefulness of Blimp1 as a germline marker. The staining was highly specific (nuclear), but only successful when using chromogenic development (used the HRP-based AEC-red system). Citrate buffer was superior to EDTA for antigen retrieval and 1:500 dilution in 0.05M Tris-HCl, pH 6.0 with 1% BSA was superior to higher concentrations and higher pH or PBS-based solution.  Detection Method: AEC red. Negative Control: no primary and isotype control. Deparaffinization: 2x10min xylene, 2x 5min 99% etOH, 3min 96% etOH, 3min 70% etOH, 3 min water. Antigen Retrieval: 0.01M citrate buffer, 15min at boiling.
  Average
ICC/IF Other 02/17/2009
Show

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedBlimp1
SpeciesOther
Other SpeciesOther
LotSS

Blocking

Blocking DetailsBlocking Buffer: BSA, Blocking Time: 1 hour, Blocking Temp: 37 degrees Celsius

Primary Anitbody

Dilution RatioPrimary Ab Incubation Dilution Buffer: 0.5% BSA, Primary Ab Dilution Ratio: 1/200, Incubation Time:1.5 hr, Incubation Temp:37C
  Average
ICC/IF Other 02/17/2009
Show

Summary

ApplicationImmunofluorescence
Sample TestedBlimp1
SpeciesOther
Other SpeciesOther
LotSS

Blocking

Blocking DetailsBlocking Buffer: BSA, Blocking Time: 1 hour, Blocking Temp: 37 degrees Celsius

Primary Anitbody

Dilution RatioPrimary Ab Incubation Dilution Buffer: 0.5% BSA, Primary Ab Dilution Ratio: 1/200, Incubation Time:1.5 hr, Incubation Temp:37C

Publications for BLIMP1/PRDM1 Antibody (NB600-235) (36)

Read what people are writing who have used BLIMP1/PRDM1 Antibody (NB600-235).

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 7 applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP.

Have you published using BLIMP1/PRDM1 Antibody (NB600-235)? Submit your review and earn rewards points which can be used for merchandise & discounts.
Filter By Application
WB
(14)
Flow
(4)
ICC/IF
(3)
IHC
(1)
IHC-Fr
(1)
IHC-P
(7)
IP
(2)
All Applications
Filter By Species
Human
(14)
Mouse
(11)
All Species
Publication Applications Species
John, S A et al. Ets-1 regulates plasma cell differentiation by interfering with the activity of the transcription factor Blimp-1. J Biol Chem;283(2):951-62. 2008 Jan 11. [PMID: 17977828] (WB, Mouse) WB Mouse
Nishikawa K, Nakashima T, Hayashi M et al. Blimp1-mediated repression of negative regulators is required for osteoclast differentiation. Proc Natl Acad Sci U S A 2010 Feb 16. [PMID: 20133620]
Maseda D, Smith SH, Dilillo DJ et al. Regulatory B10 Cells Differentiate into Antibody-Secreting Cells After Transient IL-10 Production In Vivo. Journal of Immunology 2011 Dec 23. [PMID: 22198952]
Severa, M et al. Toll-like receptor-dependent and -independent viperin gene expression and counter-regulation by PRDI-binding factor-1/BLIMP1. J Biol Chem;281(36):26188-95. 2006 Sep 8. [PMID: 16849320]
Gong, D, Malek, T R. Cytokine-dependent Blimp-1 expression in activated T cells inhibits IL-2 production. J Immunol;178(1):242-52. 2007 Jan 1. [PMID: 17182561]
Liu, Y et al. Rituximab plus CHOP (R-CHOP) overcomes PRDM1-associated resistance to chemotherapy in patients with diffuse large B-cell lymphoma. Blood;110(1):339-44. 2007 Jul 1. [PMID: 17379744] (WB, IHC-P, Human) IHC-P, WB Human
Garcia-Bates TM, Baglole CJ, Bernard MP et al. Peroxisome proliferator-activated receptor gamma ligands enhance human B cell antibody production and differentiation. J Immunol;183(11):6903-12. 2009 Dec 1. [PMID: 19915048]
Julaton VTA, Reijo Pera RA. NANOS3 function in human germ cell development. Hum Mol Genet;20(11):2238-50. 2011 Jun 1. [PMID: 21421998] (ICC/IF, Human) ICC/IF Human
Shimshon L, Michaeli A, Hadar R et al. SUMOylation of Blimp-1 promotes its proteasomal degradation. FEBS Lett;585(15):2405-9. 2011 Aug 4. [PMID: 21722636] (IP, Mouse) IP Mouse
Gao Y, Kazama H, Yonehara S. Bim regulates B-cell receptor-mediated apoptosis in the presence of CD40 signaling in CD40-pre-activated splenic B cells differentiating into plasma cells. Int Immunol. 2012 Feb 1. [PMID: 22301689]
Altin JA, Goodnow CC, Cook MC. IL-10+CTLA-4+ Th2 Inhibitory Cells Form in a Foxp3-Independent, IL-2-Dependent Manner from Th2 Effectors during Chronic Inflammation. J Immunol;188(11):5478-88. 2012 Jun 1. [PMID: 22547705] (FLOW, Mouse) Flow Mouse
Cattoretti, G et al. PRDM1/Blimp-1 is expressed in human B-lymphocytes committed to the plasma cell lineage. J Pathol;206(1):76-86. 2005 May. [PMID: 15772984] (IHC-P, WB, Human) IHC-P, WB Human
Robertson EJ, Charatsi I, Joyner CJ et al. Blimp1 regulates development of the posterior forelimb, caudal pharyngeal arches, heart and sensory vibrissae in mice. Development;134(24):4335-45. 2007 Dec. [PMID: 18039967] (ICC/IF, Mouse) ICC/IF Mouse
Courts C, Montesinos-Rongen M, Brunn A, Bug S, Siemer D, Hans V, Blumcke I, Klapper W, Schaller C, Wiestler OD, Kuppers R, Siebert R, Deckert M. Recurrent inactivation of the PRDM1 gene in primary central nervous system lymphoma. J Neuropathol Exp Neurol;67(7):720-7. 2008 Jul. [PMID: 18596541] (IHC-P, IHC-Fr, Human) IHC-Fr, IHC-P Human
Chang, DH et al. The dynamic expression pattern of B lymphocyte induced maturation protein-1 (Blimp-1) during mouse embryonic development. Mech Dev 1179(1-2): 305-309. 2002 [PMID: 12204275] (IHC-P, Mouse) IHC-P Mouse
Wang X, Belguise K, O'Neill CF et al. RelB NF-{kappa}B Represses Estrogen Receptor {alpha} Expression via Induction of the Zinc Finger Protein Blimp1. Mol Cell Biol;29(14):3832-3844. 2009 [PMID: 19433448]
Matsushita T, Tedder TF. Identifying regulatory B cells (B10 cells) that produce IL-10 in mice. Methods Mol Biol;677:99-111. 2011 [PMID: 20941605]
Le Gallou S, Caron G, Delaloy C et al. IL-2 Requirement for Human Plasma Cell Generation: Coupling Differentiation and Proliferation by Enhancing MAPK-ERK Signaling J Immunol 2012 Jul 1 [PMID: 22634617] (WB, Human) WB Human
Ramon S, Gao F, Serhan CN, Phipps RP. Specialized Proresolving Mediators Enhance Human B Cell Differentiation to Antibody-Secreting Cells J Immunol 2012 Jun 18 [PMID: 22711890] (WB, Human) WB Human
Takata K, Sato Y, Nakamura N et al. Duodenal follicular lymphoma lacks AID but expresses BACH2 and has memory B-cell characteristics Mod Pathol 2012 Aug 17 [PMID: 22899287] (IHC, Human) IHC Human
Morgan MA, Mould AW, Li L et al. Alternative splicing regulates prdm1/blimp-1 DNA binding activities and corepressor interactions Mol Cell Biol 2012 Sep [PMID: 22733990] (IP, Human) IP Human
Lee SR, Rutan JA, Monteith AJ et al. Receptor Cross-Talk Spatially Restricts p-ERK during TLR4 Stimulation of Autoreactive B Cells J Immunol 2012 Sep 14 [PMID: 22984080] (WB, Mouse) WB Mouse
Lin MH, Chou FC, Yeh LT et al. B lymphocyte-induced maturation protein 1 (BLIMP-1) attenuates autoimmune diabetes in NOD mice by suppressing Th1 and Th17 cells Diabetologia 2012 Oct 7 [PMID: 23052053] (WB, Mouse) WB Mouse
Nagy N, Adori M, Rasul A et al. Soluble factors produced by activated CD4+ T cells modulate EBV latency Proc Natl Acad Sci U S A 2012 Jan 31 [PMID: 22307606] (WB, Human) WB Human
fan CC, Lee LY, Yu MY et al. Upregulated hPuf-A promotes breast cancer tumorigenesis. Tumour Biol 2013 Apr 28 [PMID: 23625657]
Wong LY, Brulois K, Toth Z et al. KSHV K4.2 immediate early gene product regulates immunoglobulin secretion and calcium homeostasis by interacting with and inhibiting pERP1. J Virol. 2013 Aug 28 [PMID: 23986581] (IHC-P, Human) IHC-P Human
Parlato S, Bruni R, Fragapane P et al. IFN-alpha Regulates Blimp-1 Expression via miR-23a and miR-125b in Both Monocytes-Derived DC and pDC. PLoS One. 2013 Aug 16 [PMID: 23977359] (WB, Human) WB Human
Kim EH, Gasper DJ, Lee SH et al. Bach2 regulates homeostasis of Foxp3+ regulatory T cells and protects against fatal lung disease in mice. J. Immunol. 2014 Feb 1 [PMID: 24367030] (FLOW, Mouse) Flow Mouse
Severa M, Islam SA, Waggoner SN et al. The Transcriptional Repressor BLIMP1 Curbs Host Defenses by Suppressing Expression of the Chemokine CCL8. J. Immunol. 2014 Feb 24 [PMID: 24477914] (WB, Mouse) WB Mouse
Horn M, Geisen C, Cermak L et al. DRE-1/FBXO11-Dependent Degradation of BLMP-1/BLIMP-1 Governs C. elegans Developmental Timing and Maturation. Dev. Cell 3/4/2014 [PMID: 24613396] (WB, Human) WB Human
Kim J, Nakasaki M, Todorova D et al. p53 induces skin aging by depleting Blimp1(+) sebaceous gland cells. Cell Death Dis 3/28/2014 [PMID: 24675459] (IHC-P, ICC/IF, FLOW, Mouse) Flow, ICC/IF, IHC-P Mouse
Rasul E, Salamon D, Nagy N et al. The MEC1 and MEC2 Lines Represent Two CLL Subclones in Different Stages of Progression towards Prolymphocytic Leukemia. PLoS ONE. 2014 Aug 28 [PMID: 25162594] (WB, Human) WB Human
Lin MH, Yeh LT, Chen SJ et al. T cell-specific BLIMP-1 deficiency exacerbates experimental autoimmune encephalomyelitis in nonobese diabetic mice by increasing Th1 and Th17 cells. Clin Immunol. 2014 Mar 10 [PMID: 24568746] (WB, Mouse) WB Mouse
Gosal K, Dunlop K, Dhaliwal R et al. Rho-kinase Mediates Right Ventricular Systolic Dysfunction in Rats with Chronic neonatal Pulmonary Hypertension. Am. J. Respir. Cell Mol. Biol. 2014 Oct 22 [PMID: 25337652] (FLOW) Flow
Ramon S, Baker Sf, Sahler Jm et al. The Specialized Proresolving Mediator 17-HDHA Enhances the Antibody-Mediated Immune Response against Influenza Virus: A new Class of Adjuvant? J. Immunol. 2014 Nov 12 [PMID: 25392529] (WB) WB
Boy S, van Heerden M, Pool R, Willem P. Plasmablastic lymphoma versus diffuse large B cell lymphoma with plasmablastic differentiation: proposal for a novel diagnostic scoring system Journal of Hematopathology. 2015 Jan 15 (IHC-P, Human)

Details:
BLIMP1/PRDM1 antibody used for IHC-P staining on CD20-negative, ALK-negative, HHV8-negative non-Hodgkin?s human lymphomas with plasmablastic differentiation defined by their morphological features, high proliferation index and positivity for MUM1/IRF4 and PRDM/Blimp1 protein expression. IHC-P assay involved - Heat-induced epitope retrieval with TRIS/EDTA pH9 buffer, quenching of endogenous peroxidase activity with H2O2, 1:200 dilution of primary antibody, detection with Novolink Polymer Detection Kit which followed counterstaining with haematoxylin (Fig 3).
IHC-P Human

Product General Protocols

View specific protocols for BLIMP1/PRDM1 Antibody (NB600-235): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ChIP Video Protocol
ChIP Webinar
ICC/IF Video Protocol

Ask a Scientist

Submit your question on below.
During business hours, we will respond to your email within 24 hours. For any questions submitted on the weekend, a response will be received on Monday.
For immediate assistance during business hours M- F (excluding major holidays), please contact us.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.

FAQs for BLIMP1/PRDM1 (35)

  1. Does your Blimp-1 Antibody (NB600-235) recognize both the alpha and beta form of BLIMP-1
    • By WB, this antibody recognizes a band ~98 kD and may recognize one ~80 kDa (the beta form).
  2. Do you have any images (dot plot chart or histogram) of human cells stained with this ab for flow cytometry?
    • Unfortunately, I do not have any additional images for this product on my records. This is one of our best selling products with great customer feedback and citations in at least 24 peer reviewed publications in journals of high repute. We stand by the quality of our products and all our products are 100% guaranteed. We are always there to help in case you face any trouble in generating your desired outcome out of the use of our products.
  3. Do you have any references which show this works in human for IF?
    • The research publication that has evidence for IF use of the above mentioned antibody in samples of human origin is: Julaton VT, Reijo Pera RA. NANOS3 function in human germ cell development. Hum Mol Genet. 2011 Jun 1;20(11):2238-50. Epub 2011 Mar 19.
  4. May we ask if NB600-235 is suitable for ChIP?
    • NB600-235 have been validated in IP, so since BLIMP-1 is a DNA binding protein, this antibody should be suitable for ChIP. If you would be interested in testing this novel application, please take a look at our Innovator's Reward program.
  5. In the immunohistochemistry paraffin section protocol....the PBS buffer....should it be at certain pH?
    • For IHC application, different labs cites the pH of PBS buffer in the range of 7.1 - 7.6 but most commonly used pH is pH 7.4 (this is what we use in our lab). You may use this PBS (pH 7.4) for making permeablization buffer, antibody diluent buffers and for making wash buffer. For more on the protocol that we use in our lab and for IHC-P troubleshooting suggestions, you may visit: IHC-P Protocol and IHC-P Troubleshooting.
  6. I was wondering what does Format 7C mean as far as anitbodies go?
    • 7C is a fluorophore - it is useful for FLOW. The details of the fluor are (A=425, E=500)
  7. What epitope does this monoclonal antibody recognize?
    • R&D Systems does not epitope map its antibodies. The antibody is generated using the entire immunogen listed on the datasheet. If a peptide immunogen is used, this will be indicated on the datasheet.
  8. What epitope does this polyclonal antibody recognize?
    • Polyclonal antibodies generally recognize multiple epitopes because they are generated using the entire immunogen stated on the product datasheet.
  9. What are R&D Systems matched antibody pairs?
    • R&D Systems matched antibody pairs assist in the development of an ELISA. R&D Systems has demonstrated that the paired antibodies can be used in a sandwich immunoassay to recognize the recombinant protein. We recommend that the investigator have expertise in immunoassay development before attempting to use these products. Each investigator must empirically determine the optimal concentrations for the capture and detection antibodies. The amount of antibodies supplied are not normalized to develop the same number of plates. R&D Systems recombinant cytokines are not calibrated by ELISA and therefore must be mass calibrated before use. The DuoSet ELISA Development Kit product line includes the antibodies, standard, enzyme, and protocol necessary for running an immunoassay, and can often be a better value than purchasing antibody pairs and protein standard.
  10. Can I use your antibodies for a matched pair ELISA with a standard from another company?
    • Yes, but there are potential issues to consider. There can be differences in immunological recognition or the binding between antibody and recombinant protein. This can happen if there is a difference in the folding or sequence of the recombinant protein used as the standard versus the recombinant protein used as the immunogen for the antibody. Protein folding or sequence differences in the antibody-binding region can lead to poor (or no) recognition of the standard.
  11. Can I use your antibodies for a matched pair ELISA with a standard from another company?
    • Yes, but there are potential issues to consider. There can be differences in immunological recognition or the binding between antibody and recombinant protein. This can happen if there is a difference in the folding or sequence of the recombinant protein used as the standard versus the recombinant protein used as the immunogen for the antibody. Protein folding or sequence differences in the antibody-binding region can lead to poor (or no) recognition of the standard.
  12. What grade BSA is recommend for use in ELISA and ELISpot Development Systems?
    • The grade of the BSA used has been found to be a critical component for running a successful ELISA. BSA with a minimum purity of 98% is recommended. R&D Systems recommends using the same BSAs used in development of the DuoSet; Catalog # DY995 Reagent Diluent Concentrate 2, Catalog # DY997 Reagent Diluent Concentrate 1. Millipore Probumin® Diagnostic Grade BSA (Millipore, Catalog # 82-045) is another example.
  13. What grade BSA is recommend for use in ELISA and ELISpot Development Systems?
    • The grade of the BSA used has been found to be a critical component for running a successful ELISA. BSA with a minimum purity of 98% is recommended. R&D Systems recommends using the same BSAs used in development of the DuoSet; Catalog # DY995 Reagent Diluent Concentrate 2, Catalog # DY997 Reagent Diluent Concentrate 1. Millipore Probumin® Diagnostic Grade BSA (Millipore, Catalog # 82-045) is another example.
  14. What is a direct ELISA?
    • In a direct ELISA, a plate is coated with the analyte of interest and a labeled detection antibody is used to verify the presence of the analyte.The direct ELISA may use a colorimetric, chemiluminescent, or fluorescent reporter.
  15. What is a direct ELISA?
    • In a direct ELISA, a plate is coated with the analyte of interest and a labeled detection antibody is used to verify the presence of the analyte.The direct ELISA may use a colorimetric, chemiluminescent, or fluorescent reporter.
  16. What is a sandwich ELISA?
    • A sandwich ELISA uses an immobilized capture antibody specific for the analyte of interest in a sample. After the analyte is bound to the immobilized antibody, a labeled secondary antibody specific for the analyte is used for detection. The analyte is "sandwiched" between the two antibodies. The sandwich ELISA is extremely sensitive, and the values obtained are quantitative when compared with a standard curve.
  17. What is a sandwich ELISA?
    • A sandwich ELISA uses an immobilized capture antibody specific for the analyte of interest in a sample. After the analyte is bound to the immobilized antibody, a labeled secondary antibody specific for the analyte is used for detection. The analyte is "sandwiched" between the two antibodies. The sandwich ELISA is extremely sensitive, and the values obtained are quantitative when compared with a standard curve.
  18. What is the difference between AB###, AF###, BAF###, MAB###, and BAM### antibody catalog number designations?
    • AB### designated antibodies are polyclonal antibodies that are purified using Protein A, Protein G or Ion Exchange chromatography. Consequently, the resulting purified antibody is a total IgG fraction and may contain IgG not specific for the analyte of interest. AF### designated antibodies, are antigen affinity-purified and then further purified by ion exchange or size exclusion. Therefore, AF### antibodies are IgG specific only to the antigen. Antibodies that have the designation MAB### are monoclonal antibodies. BAF### and BAM### designated antibodies are the biotinylated versions of the AF### and MAB### designated antibodies, respectively.
  19. What is the difference between AB###, AF###, BAF###, MAB###, and BAM### antibody catalog number designations?
    • AB### designated antibodies are polyclonal antibodies that are purified using Protein A, Protein G or Ion Exchange chromatography. Consequently, the resulting purified antibody is a total IgG fraction and may contain IgG not specific for the analyte of interest. AF### designated antibodies, are antigen affinity-purified and then further purified by ion exchange or size exclusion. Therefore, AF### antibodies are IgG specific only to the antigen. Antibodies that have the designation MAB### are monoclonal antibodies. BAF### and BAM### designated antibodies are the biotinylated versions of the AF### and MAB### designated antibodies, respectively.
  20. What is the rationale for using trehalose to stabilize proteins?
    • Trehalose is an effective sugar for stabilizing proteins against damage caused by freezing. It can also make the protein more resistant to moisture gain when lyophilized, resulting in a product that is less likely to precipitate when reconstituted. In addition, it has been used in approved parenteral therapeutics
  21. What is the rationale for using trehalose to stabilize proteins?
    • Trehalose is an effective sugar for stabilizing proteins against damage caused by freezing. It can also make the protein more resistant to moisture gain when lyophilized, resulting in a product that is less likely to precipitate when reconstituted. In addition, it has been used in approved parenteral therapeutics
  22. Will trehalose affect my conjugation reaction?
    • It is possible that the presence of trehalose will interfere in the successful conjugation of a protein. This will depend on the method used, and the customer should investigate this potential prior to purchasing the product.
  23. Will trehalose affect my conjugation reaction?
    • It is possible that the presence of trehalose will interfere in the successful conjugation of a protein. This will depend on the method used, and the customer should investigate this potential prior to purchasing the product.
  24. Will trehalose affect the performance of the protein or antibody in my specific application?
    • We have seen no adverse effect in our bioassays or other approved applications. However, customers are advised to run a control in their assay to determine if the concentration of trelahose in the protein or antibody formulation has any adverse effects.
  25. Will trehalose affect the performance of the protein or antibody in my specific application?
    • We have seen no adverse effect in our bioassays or other approved applications. However, customers are advised to run a control in their assay to determine if the concentration of trelahose in the protein or antibody formulation has any adverse effects.
  26. Will trehalose included in the formulation affect the animal if it is injected?
    • Trehalose is unlikely to have an effect in vivo. It has been approved as an excipient for use in human injectable drugs.The trehalose used by R&D Systems is derived from Saccharomyces cerevisiae and is determined to be at minimum 98.5% pure by HPAE.
  27. Will trehalose included in the formulation affect the animal if it is injected?
    • Trehalose is unlikely to have an effect in vivo. It has been approved as an excipient for use in human injectable drugs.The trehalose used by R&D Systems is derived from Saccharomyces cerevisiae and is determined to be at minimum 98.5% pure by HPAE.
  28. What are R&D Systems matched antibody pairs?
    • R&D Systems matched antibody pairs assist in the development of an ELISA. R&D Systems has demonstrated that the paired antibodies can be used in a sandwich immunoassay to recognize the recombinant protein. We recommend that the investigator have expertise in immunoassay development before attempting to use these products. Each investigator must empirically determine the optimal concentrations for the capture and detection antibodies. The amount of antibodies supplied are not normalized to develop the same number of plates. R&D Systems recombinant cytokines are not calibrated by ELISA and therefore must be mass calibrated before use. The DuoSet ELISA Development Kit product line includes the antibodies, standard, enzyme, and protocol necessary for running an immunoassay, and can often be a better value than purchasing antibody pairs and protein standard.
  29. What is the recommended protocol for reconstituting and aliquotting lyophilized proteins and antibodies?<br>
    • Following these guidelines for reconstituting lyophilized material will help ensure complete recovery of the protein or antibody.

      1) Tap or briefly centrifuge the vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.

      2) Use the buffer and stock concentration recommended in the product datasheet. If you want a stock concentration that is higher than the one recommended, contact Technical Service for specific recommendations.

      3) For optimal recovery, reconstitute using room temperature buffer.

      4) After adding the buffer, re-cap the vial and invert gently by hand or place on a slow rocking platform. This will allow the reconstitution buffer to coat all the surfaces inside the vial. Do not mix by vortexing or by pipetting the material up and down.

      5) Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.

      6) Store the reconstituted protein in polypropylene or siliconized tubes. If aliquotting, it is recommended that aliquots be no smaller than 10 μL. In addition, avoid repeated freeze/thaw cycles.

  30. What is the recommended protocol for reconstituting and aliquotting lyophilized proteins and antibodies?<br>
    • Following these guidelines for reconstituting lyophilized material will help ensure complete recovery of the protein or antibody.

      1) Tap or briefly centrifuge the vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.

      2) Use the buffer and stock concentration recommended in the product datasheet. If you want a stock concentration that is higher than the one recommended, contact Technical Service for specific recommendations.

      3) For optimal recovery, reconstitute using room temperature buffer.

      4) After adding the buffer, re-cap the vial and invert gently by hand or place on a slow rocking platform. This will allow the reconstitution buffer to coat all the surfaces inside the vial. Do not mix by vortexing or by pipetting the material up and down.

      5) Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.

      6) Store the reconstituted protein in polypropylene or siliconized tubes. If aliquotting, it is recommended that aliquots be no smaller than 10 μL. In addition, avoid repeated freeze/thaw cycles.

  31. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  32. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  33. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  34. Has a product ever been used for an application that is not listed on the datasheet?
    • If a specific application is not listed on the datasheet, it may mean that this product has not been tested in this application, or it may mean that in-house testing in this application did not meet R&D Systems’specifications. Please check our Citations tab to see if other researchers have published using your application, sample type and/or species. If you would like more information on whether or not an application has been tested, please contact Technical Service at (800) 343-7475.

  35. Has a product ever been used for an application that is not listed on the datasheet?
    • If a specific application is not listed on the datasheet, it may mean that this product has not been tested in this application, or it may mean that in-house testing in this application did not meet R&D Systems’specifications. Please check our Citations tab to see if other researchers have published using your application, sample type and/or species. If you would like more information on whether or not an application has been tested, please contact Technical Service at (800) 343-7475.