BLIMP1/PRDM1 Antibody (3H2-E8)

Western Blot: BLIMP1/PRDM1 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.

Immunocytochemistry/Immunofluorescence: BLIMP1/PRDM1 Antibody (3H2-E8) [NB600-235] - A431 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were incubated with anti-BLIMP1 (3H2-E8) at 10 ug/ml overnight at 4C and detected with an anti-mouse IgG Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: BLIMP1/PRDM1 Antibody (3H2-E8) [NB600-235] - Analysis using paraffin-embedded human tonsil tissue with BLIMP1/PRDM1 antibody at dilution of 1:50. Detection is completed through VC001 (DAB) and Counterstained by hematoxylin; labeling is predominantly in germinal centers. Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.

Flow Cytometry: BLIMP1/PRDM1 Antibody (3H2-E8) [NB600-235] - An intracellular stain was performed on U266 cells with BLIMP1/PRDM1 Antibody (3H2-E8) NB600-235 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and permeabilized with 0.1% Saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems).

Immunocytochemistry/Immunofluorescence: BLIMP1/PRDM1 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.

Immunocytochemistry/Immunofluorescence: BLIMP1/PRDM1 Antibody (3H2-E8) [NB600-235] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-BLIMP1/PRDM1 (3H2-E8) conjugated to DyLight 550 [NB600-235R] at 20ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow Cytometry: BLIMP1/PRDM1 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).

Flow Cytometry: BLIMP1/PRDM1 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)

Flow Cytometry: BLIMP1/PRDM1 Antibody (3H2-E8) [NB600-235] - Blimp-1 expression by CD19+ (thick black line) or CD8+ (thin, shaded line). (Tedder Lab; Duke Univ).

Flow Cytometry: BLIMP1/PRDM1 Antibody (3H2-E8) [NB600-235] - Analysis using the Biotin conjugate of NB600-235. Staining of Blimp-1 in mouse spleen. Image courtesy of product review by Branislav Krljanac.

• Reactivity: Hu, Mu, Po
• Applications: WB, ChIP, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP
• Clone: 3H2-E8
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

BLIMP1/PRDM1 Antibody (3H2-E8) Summary

• Immunogen: A fragment of mouse Blimp-1 corresponding to residues 199-409. [UniProt# Q60636]
• Localization: Nuclear
• Marker: Plasma Cell Marker
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: PRDM1
• Purity: Protein G purified

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 Control:

  BLIMP1/PRDM1 Lysate (NBL1-14733)

Reactivity Notes

Human, mouse and pig.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine and 0.15M NaCl
• Preservative: 0.1% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

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
• BLIMP1
• BLIMP-1
• BLIMP1MGC118925
• blmp-1
• 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
• PRDM1
• PR-domain zinc finger protein 1
• ZNFPR1A1

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.

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.

Publications for BLIMP1/PRDM1 Antibody (NB600-235)(42)

Publications using NB600-235

• Wu L, Ehlin-Henriksson B, Zhou X et al. EBV provides survival factors to EBV+ DLBCL lines and modulates cytokine induced specific chemotaxis in EBV+ DLBCL Immunology 2017 Jul 12 [PMID: 28699226] (Human)
• McCarthy C, Kenny LC. Therapeutically targeting mitochondrial redox signalling alleviates endothelial dysfunction in preeclampsia Sci Rep Sep 8 2016 [PMID: 27604418] (ICC/IF, Human)
• Jang KJ, Mano H, Aoki K et al. Mitochondrial function provides instructive signals for activation-induced B-cell fates Nat Commun 2015 Apr 10 [PMID: 25857523] (WB, FLOW, Mouse)
• 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)
• 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)
• Joo H, Li D, Dullaers M et al. C-Type Lectin-like Receptor LOX-1 Promotes Dendritic Cell-Mediated Class-Switched B Cell Responses. Immunity 2014 Oct 08 [PMID: 25308333] (FLOW, Human)
  
  Details:
  Using the PE conjugated version of NB600-235, catalog number NB600-235PE.
• 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)
• 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)
• 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)
• 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)
• 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)
• 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)
• 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)
• 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)
• fan CC, Lee LY, Yu MY et al. Upregulated hPuf-A promotes breast cancer tumorigenesis. Tumour Biol 2013 Apr 28 [PMID: 23625657]
• 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)
• 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)
• 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)
• 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)
• 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)
• 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)
• 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).
• 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)
• 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)
• 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]
• Hussaini M, Yeung C, Zhu X et al. PRDM1 expression levels in marginal zone lymphoma and lymphoplasmacytic lymphoma Int J Clin Exp Patho 2017 Aug 15 (IHC-P, Human)
• 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]
• 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)
• 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)
• 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]
• 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]
• Brzezinski JA 4th, Lamba DA, Reh TA. Blimp1 controls photoreceptor versus bipolar cell fate choice during retinal development. Development 2010 Feb [PMID: 20110327]
• 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]
• 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]
• 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)
• 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)
• 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)
• 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)
• 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]
• 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]
• 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)
• 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)

Reviews for BLIMP1/PRDM1 Antibody (NB600-235) (2)

Average Rating: 3.5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Other.

reviewed by: Anonymous IHC-P Other 05/31/2010

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Porcine embryo
• Species: Other
• Comments: I 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 Details: Blocking Buffer: 2% BSA, Blocking Time: 10 minutes, Blocking Temp: room temperature

Primary Anitbody

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

Secondary Antibody

• Secondary Description: Secondary Ab: DAKO LSAB+ kit (SA-HRP link)

Details

• Detection Notes: Please see comments
• Fixation Details: PFA fixed and paraffin embedded, Deparaffinization: 2x 10min xylene, 2x 5min 99% etOH, 3min 96% etOH, 3min 70% etOH, 3min water

Comments

• Comments: I 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.

reviewed by: Anonymous IF Other 02/17/2009

Summary

• Application: Immunofluorescence
• Sample Tested: Blimp1
• Species: Other
• Lot: SS

Blocking

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

Primary Anitbody

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

Secondary Antibody

• Secondary Description: Secondary Ab: Alexa 594

FAQs for BLIMP1/PRDM1 Antibody (NB600-235). (Showing 1 - 4 of 4 FAQs).

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.

Bioinformatics

• Gene Symbol: PRDM1
• Entrez:
  • 639(Human)
  • 12142(Mouse)
  • 309871(Rat)