14-0194-37

antibody from Invitrogen Antibodies

Targeting: CD19

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

14-0194-37

Provider

Invitrogen Antibodies

Product name

CD19 Monoclonal Antibody (6OMP31), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The 6OMP31 monoclonal antibody reacts with human, mouse, and rat CD19. CD19 is a transmembrane glycoprotein belonging to the immunoglobulin superfamily. CD19 was first identified as the B4 antigen of human B lymphocytes using the anti-B4 monoclonal. It is expressed by B cells during all stages of development, except on terminally differentiated plasma cells, and on follicular dendritic cells. CD19 acts primarily as a B cell co-receptor and together with CD21, CD81, Leu13, and MHC class II forms a multi-molecular complex that associates with the BCR. CD19 plays a role in the development, activation, differentiation, and proliferation of B cells. CD19 deficiency leads to an overall impaired humoral immune response. Mutations in CD19 have been associated with severe immunodeficiency syndromes. Although it is unclear whether CD19 contributes directly to B cell carcinogenesis, it is expressed on most B cell tumors, such as acute lymphoblastic leukemias and B cell lymphomas. The use of CD19 monoclonal antibodies for therapies against lymphoma, leukemia and autoimmune disorders are currently being explored. Applications Reported: This 6OMP31 antibody has been reported for use in western blotting, immunohistochemical staining of frozen tissue sections, and immunohistochemical staining of formalin-fixed paraffin embedded tissue sections. Applications Tested: This 6OMP31 antibody has been tested by immunohistochemistry of formalin-fixed paraffin embedded tissue using low or high pH antigen retrieval and can be used at less than or equal to 0.5 µg/mL. The 6OMP31 antibody has been tested by immunohistochemstry of frozen tissue and can be used at less than or equal to 0.5 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.

Reactivity

Human, Mouse, Rat

Host

Rat

Isotype

IgG

Antibody clone number

6OMP31

Vial size

2 mg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

Knockdown of CD146 promotes endothelial-to-mesenchymal transition via Wnt/β-catenin pathway.

Zhang ZY, Zhai C, Yang XY, Li HB, Wu LL, Li L
PloS one 2022;17(8):e0273542

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Fecal microbiota transplantation ameliorates atherosclerosis in mice with C1q/TNF-related protein 9 genetic deficiency.

Kim ES, Yoon BH, Lee SM, Choi M, Kim EH, Lee BW, Kim SY, Pack CG, Sung YH, Baek IJ, Jung CH, Kim TB, Jeong JY, Ha CH
Experimental & molecular medicine 2022 Feb;54(2):103-114

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Distribution of Immune Cells Including Macrophages in the Human Cochlea.

Liu W, Danckwardt-Lillieström N, Schrott-Fischer A, Glueckert R, Rask-Andersen H
Frontiers in neurology 2021;12:781702

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Innate-like self-reactive B cells infiltrate human renal allografts during transplant rejection.

Asano Y, Daccache J, Jain D, Ko K, Kinloch A, Veselits M, Wolfgeher D, Chang A, Josephson M, Cunningham P, Tambur A, Khan AA, Pillai S, Chong AS, Clark MR
Nature communications 2021 Jul 16;12(1):4372

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Single-cell evaluation reveals shifts in the tumor-immune niches that shape and maintain aggressive lesions in the breast.

Sinha VC, Rinkenbaugh AL, Xu M, Zhou X, Zhang X, Jeter-Jones S, Shao J, Qi Y, Zebala JA, Maeda DY, McAllister F, Piwnica-Worms H
Nature communications 2021 Aug 18;12(1):5024

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Kynurenine plays an immunosuppressive role in 2,4,6-trinitrobenzene sulfate-induced colitis in mice.

Tashita C, Hoshi M, Hirata A, Nakamoto K, Ando T, Hattori T, Yamamoto Y, Tezuka H, Tomita H, Hara A, Saito K
World journal of gastroenterology 2020 Mar 7;26(9):918-932

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Microsomal Prostaglandin E Synthase-1 Facilitates an Intercellular Interaction between CD4⁺ T Cells through IL-1β Autocrine Function in Experimental Autoimmune Encephalomyelitis.

Takemiya T, Takeuchi C, Kawakami M
International journal of molecular sciences 2017 Dec 19;18(12)

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot was performed using Anti-CD19 Monoclonal Antibody (6OMP31), eBioscience™ (Product # 14-0194-80, 14-0194-82) and a 80 kDa band corresponding to B-lymphocyte antigen CD19 was observed across cell lines tested. Membrane enriched extracts (30 µg lysate) of Raji (Lane 1), Ramos (Lane 2), Daudi (Lane 3), SW480 (Lane 4), Jurkat (Lane 5) and MOLT-4 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with F(ab2-Rabbit anti-Rat IgG (H+L Secondary Antibody, HRP (Product # PA1-29927, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockout of CD19 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR807388_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of CD19 was performed by loading 30 µg of Raji Cas9 (Lane 1) and Raji CD19 KO (Lane 2) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with CD19 Monoclonal Antibody (6OMP31), eBioscience™ (Product # 14-0194-82, 1:500 dilution) and F(ab)2-Rabbit anti-Rat IgG (H+L) Secondary Antibody, HRP (Product # PA1-29927, 1:5000 dilution) using the iBright™ FL 1500 (Product # A44115). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to CD19.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of B-lymphocyte antigen CD19 was performed using 70% confluent log phase Ramos cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CD19 Monoclonal Antibody (6OMP31), eBioscience™ (Product # 14-0194-80, 14-0194-82) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A11006), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing membrane localization. Panel e represents no expression of CD19 in Jurkat cells .Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry of formalin-fixed paraffin embedded human tonsil using 0.5 µg/mL Rat IgG2a K Isotype Control Purified (left) or 0.5 µg/mL Anti-CD19 Purified (right) followed by Anti-Rat IgG Biotin, Streptavidin HRP and DAB visualization. Nuclei are counterstained with hematoxylin.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 6 The aggressive niche harbors an expanded and spatially restricted myeloid population. a - c UMAP was used for dimension reduction of single-cell data. Immune fractions from indolent and aggressive lesions were computationally merged ( a ) for cluster analysis and identification ( b ). Proportion of clusters per group shown in c . NK natural killer, pDC plasmacytoid dendritic cell, cDC conventional dendritic cell. d Heatmap of top differentially expressed (DE) genes for each cluster, ordered in decreasing cluster size. Clusters indicated by top horizontal bar, with inferred immune identity noted. Cells derived from indolent or aggressive tumor sample indicated by second horizontal bar. DE genes are listed on the left. e Immunohistochemical staining for major lymphoid (T cells--CD3; B cells--CD19) and myeloid (macrophages--F4/80; neutrophils--MPO) populations associated with indolent and aggressive lesions. Arrows point to selected positively stained cells. Representative images shown from n = 4, 3, 3, 4 animals for CD3, CD19, F4/80, and MPO, respectively. f Immunofluorescent staining for neutrophils using an alternative marker S100A8. Left panel shows low power magnification view of adjacent indolent and aggressive lesions, with S100A8+ infiltration tightly localized with the aggressive niche. Boxes correspond by color to high magnification images shown middle (blue, indolent) and right (yellow, aggressive) panels. Representative images shown from n = 7 animals. g Immun

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 13 (A) CD19-positive B lymphocyte in the modiolus. The cell also expresses MHCII. (B) CD4 and CD8-positive T cells in the peripheral region of the RC. (C) A CD8-positive lymphocyte and an MHCII-expressing cell in Rosenthal's canal. (D) A CD8 cell is seen to physically interact with an IBA1-positive macrophage.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

The accumulation of immune cells results in atherosclerosis progression. a , b Immunofluorescence staining for the lymphocyte marker CD45 (red) and DAPI-stained nuclei (blue) are shown. c , d T cells were stained for CD3. e , f B cells were stained for CD19. g , h Macrophages were stained for CD68. * p < 0.05 for the WT-FMT group and the KO group compared to the WT group, # p < 0.05 for the KO-FMT group compared to the KO group. Two-way comparisons were performed with t tests. Scale bars in right lower corner represent 50 mum.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

S2 Fig CD146 was not expressed in SMCs and certain immune cells in adult mouse hearts. Representative double-immunofluorescence images of CD146 with alpha-SMA (a marker for SMCs), MPO (a marker for neutrophils), CD68 (a marker for monocytes/macrophages), CD19 (a marker for B lymphocytes), and CD4 (a marker for T-helper cells) in left ventricles from adult mice (scale bars indicate 50mum). Green represents CD146; red represents CD4, CD19, CD68, MPO and alpha-SMA, respectively; blue represents nuclei. (TIF) Click here for additional data file.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1 Increased number of F4/80 + KMO + cells in the colon exacerbates TNBS-induced colitis. Mice were treated with either TNBS or vehicle, and colons were collected three days after treatment. A: Histological scores determined based on a histological grading as described in Materials and Methods. B: Representative images of H&E-stained colons of the said mice. C, D: The KMO + cells in the colons of the said mice were counted using immunohistochemical staining for KMO. E: KMO mRNA expression in the colon was determined by quantitative PCR. F: The colons of TNBS-treated mice were stained for F4/80, CD19, CD11c, KMO, and DAPI (nuclei) using immunofluorescence staining. The negative controls for F4/80, CD19, CD11c, and KMO were not stained ( Supplementary Figure 2 and 3 ). a P < 0.05 vs Vehicle group, b P < 0.01 vs Vehicle group, c P < 0.05 vs TNBS-treated KMO +/+ mice group. KMO: Kynurenine 3-monooxygenase; TNBS: Trinitrobenzene sulfonic acid.