Invitrogen

Blimp-1 Monoclonal Antibody (6D3), eBioscience™

Product Image Gallery

Blimp-1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry on formalin-fixed paraffin embedded human tonsil tissue with citrate buffer antigen retrieval, using 10 µg/mL of Anti-Human/Mouse Blimp1 Purified or 10 µg/mL of Rat IgG2a kappa Isotype Control Purified (Product # 14-4321-82) followed by Anti-Rat IgG Biotin, and DAB visualization.

Blimp-1 Antibody in Western Blot (WB) — Immunoblotting of reduced cell lysates prepared from A20 cell line using 5 µg/mL of Anti-Human/Mouse Blimp1 Purified. Bands were visualized using Anti-Rat IgG HRP.

Blimp-1 Antibody in Immunocytochemistry (ICC/IF) — Figure 2 Similar gene expression profile and germ cell-specific protein level of hPGCLCs generated via 0.35% MC method and U96 method. (A) Quantitative analysis of the expression of specific genes in EpCAM-/INTEGRINalpha6-high cells (day 4) via U96 method and 0.35% MC method, respectively. The error bars represent mean +- SD with three biological replicates (white, iMeLC; green, U96; red, MC). (B) Immunofluorescence analysis of OCT4, TFAP2C, BLIMP1, and SOX17 on day 4 embryoids from 0.35% MC method and U96 method. The nuclei were stained with Hoechst (left). Scale bars, 20 mum. (C) The proportion of Sox17 (+) cells at day 4 EBs via 0.35% MC method and U96 method (right); gray, U96; dark green, 0.35% MC; ** P < 0.01.

Blimp-1 Antibody in Immunocytochemistry, Immunohistochemistry (ICC/IF, IHC) — Extended Data Fig.8 Effect of NANOG on hPGC induction, characterization of NANOS3-tdTomato reporter hPSC containing inducible SOX17, BLIMP1 with or without TFAP2C, and similarity between Cytokine- and SOX17/BLIMP1-induced hPGC a. Represents overexpression of Dex-inducible NANOG transgenes in NT reporter Comp-hPSC. b. Day4 embryoids following induction of NANOG (by Dex), with or without cytokines as indicated. c. FACS patterns after induction of hPGCs; NT/AP +ve cells (%) shown in Extended Data Fig. 8b. d. Represents overexpression of Dex-inducible SOX17, Dox-inducible BLIMP1, Shield1(S1)-inducible TFAP2C transgenes in NT reporter Comp-hPSC. e. Immunostaining of NT reporter Comp-hPSC +iSBT 1 day after induction of SOX17, BLIMP1 and TFAP2C by addition of Dex, Dox or S1. Scale bar: 50 um. f. Immunostaining of NT reporter Comp-hPSC +iSB 1 day after induction of SOX17 or BLIMP1 by addition of Dex or Dox. Scale bar: 50 um. g . Immunostaining of day 2 embyroid induced with or without Dex to induce nuclear localization of SOX17. Notably, accumulation of SOX17 signal is observed in +Dex condition. h. Changes in gene expression (RT-qPCR) during hPGC induction: Comp-hPSCs control (AP +ve cells); NT +ve hPGCs induced by SOX17/BLIMP1 or Cytokines; NT -ve cells in cells exposed to Cytokines. i. Unsupervised hierarchical clustering (UHC) of gene expression. j. Gene set enrichment analysis (GSEA) of 123 hPGC-specific genes ( Supplementary Table 1 ) on the transcriptome o

Blimp-1 Antibody in Immunocytochemistry, Immunohistochemistry (Frozen) (ICC/IF, IHC (F)) — Figure 3 CD38 Expression in Human Germ-Cell-Related Cells and Epigenetic Changes in hPGCLCs (A) FACS analysis of NANOS3-mCherry and CD38 on WIS2-NANOS3-mCherry cell line cultured in 4i medium and on day 4 and 5 embryoids following hPGCLC induction. Ratios of CD38 low and high expression in the NANOS3-mCherry-positive cells are indicated. (B) FACS histogram of CD38 low and high populations in TCam-2. (C) FACS analysis of CD38 and TNAP on genital ridges isolated from a week 6 human embryo. (D) Expression analysis by RT-qPCR for FACS-sorted TNAP-positive 4i hESCs (TNAP+ hESC) and CD38 low or high/NANOS3-mCherry day 5 hPGCLCs. Relative expression levels are shown with normalization to beta-ACTIN . Error bars indicate mean +- SD from two independent biological replicates. (E and F) Immunofluorescence analysis for 5hmC (E) and TET1 (F) on day 4 embryoids cryosection. OCT4 or BLIMP1 were used to identify hPGCLCs (highlighted). Scale bars, 50 mum. (G) Quantification of immunofluorescence intensity of various epigenetic marks/modifiers in hPGCLCs and somatic neighbors in day 1-4 embryoids (see also Figures S3 A-S3C). For UHRF1, only KI-67-positive (proliferating) cells were used for quantification. Numbers below each box denotes number of cells analyzed. Black central line represents the median, boxes and whiskers represent the 25 th and 75 th , and 2.5 th and 97.5 th percentiles, respectively. Wilcoxon signed-rank test was used to test for statistical significance. #p < 0.05; * p < 0

Blimp-1 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Figure S5 Generation of BLIMP1 KO in WIS2-NANOS3-mCherry hESC Line, Related to Figure 5 (A) Targeting strategy of BLIMP1 knockout in hESC with the designated guide RNA (gRNA) and the resulting deleted sequences. (B) Immunofluorescence of SOX17, NANOS3-mCherry and OCT4 (upper panel); and BLIMP1, NANOG and TFAP2C on wild-type (WT) and BLIMP1 KO day 4 embryoids. Scale bars = 70 mum.

Blimp-1 Antibody in Immunohistochemistry (IHC) — Fig. 4. Macrophage-derived MCPIP1 regulates B-lymphocyte expansion in lymphoid organs. (A-D) Spleen sections were stained with anti-mouse IRF4, BLIMP1, CD95 and B220 antibodies and quantified by Adobe Photoshop software as percentage of positively stained high-power field (HPF) from eight mice per group; *** P <0.001. (magnification x200, A,B; x400, C; x100, D) (E) The proliferative activity of primary B lymphocytes stimulated with serum from wild-type and knockout mice. Data are means+-s.e.m. (F) The proliferative activity of A20 B-lymphocyte cell line stimulated with serum from wild-type and knockout mice and CpG. Data are means+-s.e.m. ; *** P <0.001.

Blimp-1 Antibody in Western Blot, Immunohistochemistry (WB, IHC) — Fig 2 The eGFP-tagged Blimp1-fusion protein is efficiently expressed in the developing placenta and embryonic small intestine, but fails to rescue germ cell defects. (A) Immunohistochemical staining of E9.5 placentae demonstrates the eGFP-tagged Blimp1-fusion protein is correctly expressed in the spongiotrophoblast layer. The position of the central maternal artery is outlined. Bar, 200 mum. SpT, spongiotrophoblast; dec, maternal decidua; lab, labyrinth trophoblast. (B) Western blot analysis demonstrates robust expression in E16.5 small intestine. RIPA lysates of STO fibroblasts are included as a negative control. Levels of Blimp1 expression were quantified relative to the first wild type littermate. (C) Immunohistochemical staining demonstrates nuclear expression of both endogenous and eGFP-tagged Blimp1 protein in E16.5 villus epithelium. Bar, 200 mum. (D) The eGFP-tagged Blimp1 knock-in allele fails to reconstitute germ cell formation. Fast red alkaline phosphatase staining at E7.5 demonstrates markedly reduced numbers of PGCs (black arrows). Hematoxylin- and eosin-stained sections confirm that adult homozygous BEG/BEG testes lack spermatocytes. Bar, 200 mum.

Blimp-1 Antibody in Flow Cytometry (Flow) — Extended Data Fig.9 Response of SOX17 KO Comp-hPSC to SOX17 a. Overexpression of Dex-inducible SOX17 (iS) in SOX17 KO Comp-hPSC. b. Gene expression (RT-qPCR) on day 4 of FACS-sorted NANOS3-mCherry (NC)/Alkaline phosphatase (AP) +ve hPGCs by RT-qPCR. c. FACS analysis of d2 embryoids induced from WT Comp-hPSC, SOX17 KO Comp-hPSC and SOX17KO Comp-hPSC rescued with SOX17GR transgene (iS) (% CXCR4/AP+ cells). d. FACS pattern of day 2 embyoid induced from NT reporter Comp-hPSC showing AP/CXCR4 +ve cells expressing NT. e. Represents SOX17 inducible system (iSdd). Expression of SOX17 fused with destabilized domain (DD) can be induced by Doxycycline (Dox); addition of Shield1 (S1) can stabilize SOX17-DD protein. f. Western blots showing SOX17 expression level in day 5 embryoids from SOX17KO+iSdd Comp-hPSC. Embryoids were induced with Cytokines. To induce SOX17, different concentration of Dox and S1 were added. As controls, NC/AP +ve hPGCs and NC/AP -ve cells from WT Comp-hPSC-derived embryoids induced with Cytokines were used. Histone H3 (H3) was used for internal control. g. Immunostaining of day 4 embryoids from SOX17KO+iSdd Comp-hPSC. Embryoids from SOX17 KO and WT Comp-hPSC induced with Cytokines were used as controls. Scale bar: 50 mum. h. Quantification of immunostaining data in Extended Data Fig. 9g. The numbers of OCT4/BLIMP1 +ve hPGCs, FOXA2 +ve endodermal cells and OCT4/BLIMP1 +ve hPGCs expressing FOXA2 were counted from 3 different embryoids. The proportions of the 3 popula

Blimp-1 Antibody in ChIP assay (ChIP) — Published figure using Blimp-1 monoclonal antibody (Product # 14-5963-82) in ChIP assay

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1-10 µg/mL

View 7 publications 7 publications

Immunohistochemistry (IHC)

View 8 publications 8 publications

Immunohistochemistry (Paraffin) (IHC (P))

10 µg/mL

View 2 publications 2 publications

Immunohistochemistry (Frozen) (IHC (F))

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

View 4 publications 4 publications

Flow Cytometry (Flow)

View 1 publication 1 publication

ChIP assay (ChIP)

View 2 publications 2 publications

Product Specifications

Species Reactivity

Human, Mouse

Published species

Human, Mouse

Host/Isotype

Rat / IgG2a

Class

Monoclonal

Type

Antibody

Clone

6D3

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_1907437

Product Specific Information

Description: This 6D3 monoclonal antibody reacts with B lymphocyte-induced maturation protein-1 (Blimp-1), a transcriptional repressor expressed in B and T cells. This approximately 90-kDa protein contains five consecutive Kruppel-like zinc finger domains by which it binds DNA. In B cells, Blimp-1 promotes terminal differentiation into antibody-secreting plasma cells during a primary immune response. In addition, Blimp-1 has been implicated in germinal center formation in response to T-dependent and T-independent antigens, as well as in some diffuse large B cell lymphomas. Expressed in CD4+ and CD8+ effector/memory T cells and FoxP3+ regulatory T cells, reports have suggested possible roles for Blimp-1 in T cell homeostasis and suppression of IL-2 production. Moreover, by suppressing Th1 differentiation, Blimp-1 plays a role in T helper cell differentiation. Finally, Blimp-1 has been demonstrated to play a significant role in the terminal differentiation of effector and effector memory CD8+ T cells during viral infection.

Applications Reported: This 6D3 antibody has been reported for use in immunoblotting (WB) and immunohistology staining of paraffin-embedded tissue sections.

Applications Tested: This 6D3 antibody has been tested by western blot of reduced A20 cell lysates. For western blottiing, this antibody can be used at 1-10 µg/mL. Moreover, this antibody was tested by immunohistochemistry of formalin-fixed paraffin embedded human tonsil tissue. Antigen retrieval was performed using 10mM citrate buffer at pH 6.0. For immunohistochemistry, this antibody can be used at 10 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Blimp-1 (B lymphocyte maturation protein-1) is a nuclear protein with five zinc fingers in its carboxy terminus. It is a transcriptional repressor that is both required and sufficient to trigger terminal differentiation of B lymphocytes and monocyte/macrophages. Blimp-1 is expressed in, and may be required for terminally differentiated epithelial cells, retinal neurons and other lineage cells. It has also been implicated in the repression of beta-interferon (IFN-beta) and c-myc gene expression.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: B lymphocyte-induced maturation protein 1; B-lymphocyte-induced maturation protein 1; Beta-interferon gene positive regulatory domain I-binding factor; beta-interferon gene positive-regulatory domain I binding factor; BLIMP 1; BLIMP-1; MGC118922; MGC118923; MGC118924; MGC118925; OTTHUMP00000016918; OTTHUMP00000202173; 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 BF1; PRDI-BF1; PRDI-binding factor-1; RP1-134E15.1

Gene Aliases: b2b1765Clo; Blimp-1; BLIMP1; PRDI-BF1; PRDM1; ZNFPR1A1

UniProt ID: (Human) O75626, (Mouse) Q60636

Entrez Gene ID: (Human) 639, (Mouse) 12142

Molecular Function: C2H2 zinc finger transcription factor