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  1. Link

    cd276-antibody-sp206-ab227670.pdf

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Immunology Immune Interventions Transplantation
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD276 antibody [SP206] (ab227670)

  • Datasheet
  • SDS
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Western blot - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Western blot - Anti-CD276 antibody [SP206] (ab227670)
  • Western blot - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
  • Anti-CD276 antibody [SP206] (ab227670)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP206] to CD276
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Protein
Recombinant human CD276 protein (Fc Chimera Active) (ab216204)
Knockout
Product image
Human CD276 knockout HeLa cell line (ab261756)

View more associated products

Overview

  • Product name

    Anti-CD276 antibody [SP206]
    See all CD276 primary antibodies
  • Description

    Rabbit monoclonal [SP206] to CD276
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, LNCaP and HEK-293T cell lysate. IHC-P: Human tonsil, testis, stomach, stomach adenocarcinoma, lung, lung adenocarcinoma, lung SSCa, colon, colon adenocarcinoma, breast and breast ductal carcinoma tissues.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.1% Sodium azide
    Constituents: 1% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    SP206
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Immune Interventions
    • Transplantation
    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Cancer
    • Tumor biomarkers
    • Other
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-CD276 antibody [SP206], prediluted (ab228178)
    • Anti-CD276 antibody [SP206] - BSA and Azide free (ab240407)
    • Anti-CD276 antibody [SP206] - Low endotoxin, Azide free (ab246794)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CD276 knockout HeLa cell line (ab261756)
    • Human CD276 knockout HEK-293T cell line (ab266658)
  • KO cell lysates

    • Human CD276 knockout HEK-293T cell lysate (ab257097)
    • Human CD276 knockout HeLa cell lysate (ab257098)
  • Recombinant Protein

    • Recombinant human CD276 protein (Fc Chimera Active) (ab216204)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab227670 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
1/400. Predicted molecular weight: 57 kDa.

Primary antibody incubation for 1 hour at room temperature.

IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Primary antibody incubation for 10 minutes at room temperature.

Notes
WB
1/400. Predicted molecular weight: 57 kDa.

Primary antibody incubation for 1 hour at room temperature.

IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Primary antibody incubation for 10 minutes at room temperature.

Target

  • Function

    May participate in the regulation of T-cell-mediated immune response. May play a protective role in tumor cells by inhibiting natural-killer mediated cell lysis as well as a role of marker for detection of neuroblastoma cells. May be involved in the development of acute and chronic transplant rejection and in the regulation of lymphocytic activity at mucosal surfaces. Could also play a key role in providing the placenta and fetus with a suitable immunological environment throughout pregnancy. Both isoform 1 and isoform 2 appear to be redundant in their ability to modulate CD4 T-cell responses. Isoform 2 is shown to enhance the induction of cytotoxic T-cells and selectively stimulates interferon gamma production in the presence of T-cell receptor signaling.
  • Tissue specificity

    Ubiquitous but not detectable in peripheral blood lymphocytes or granulocytes. Weakly expressed in resting monocytes. Expressed in dendritic cells derived from monocytes. Expressed in epithelial cells of sinonasal tissue. Expressed in extravillous trophoblast cells and Hofbauer cells of the first trimester placenta and term placenta.
  • Sequence similarities

    Belongs to the immunoglobulin superfamily. BTN/MOG family.
    Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 2 Ig-like V-type (immunoglobulin-like) domains.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession Q5ZPR3 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 80381 Human
    • Entrez Gene: 102657 Mouse
    • Entrez Gene: 315716 Rat
    • Omim: 605715 Human
    • SwissProt: Q5ZPR3 Human
    • SwissProt: Q8VE98 Mouse
    • SwissProt: Q7TPB4 Rat
    • Unigene: 77873 Human
    • Unigene: 5356 Mouse
    • Unigene: 76129 Rat
    see all
  • Alternative names

    • 4Ig B7 H3 antibody
    • 4Ig-B7-H3 antibody
    • AU016588 antibody
    • B7 H3 antibody
    • B7 homolog 3 antibody
    • B7-H3 antibody
    • B7H3 antibody
    • B7RP-2 antibody
    • CD_antigen=CD276 antibody
    • CD276 antibody
    • CD276 antigen antibody
    • CD276 molecule antibody
    • CD276_HUMAN antibody
    • Costimulatory molecule antibody
    • Flags: Precursor antibody
    • PSEC0249 antibody
    • UNQ309/PRO352 antibody
    see all

Images

  • Western blot - Anti-CD276 antibody [SP206] (ab227670)
    Western blot - Anti-CD276 antibody [SP206] (ab227670)
    All lanes : Anti-CD276 antibody [SP206] (ab227670) at 1/400 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : CD276 knockout HeLa cell lysate
    Lane 3 : LNCaP cell lysate
    Lane 4 : Raji cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 57 kDa
    Observed band size: 90-110 kDa why is the actual band size different from the predicted?



    Lanes 1- 4: Merged signal (red and green). Green - ab227670 observed at 90-110 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab227670 was shown to react with CD276 antigen in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261756 (knockout cell lysate ab257098) was used. Wild-type HeLa and CD276 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab227670 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 400 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling CD276 with ab227670 at 1/100 dilution (2.29 µg/ml). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Mainly membranous staining on the human lung carcinoma, performed on a Leica Biosystems BOND™ RX instrument.
    The section was incubated with ab227670 for 10 mins at room temperature.

  • Western blot - Anti-CD276 antibody [SP206] (ab227670)
    Western blot - Anti-CD276 antibody [SP206] (ab227670)
    All lanes : Anti-CD276 antibody [SP206] (ab227670) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : CD276 knockout HEK293T cell lysate
    Lane 3 : LNCaP cell lysate
    Lane 4 : Raji cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 57 kDa
    Observed band size: 90-110 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab227670 observed at 90-110 kDa. Red - loading control ab8245 observed at 36 kDa.

    ab227670 Anti-CD276 antibody [SP206] was shown to specifically react with CD276 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266658 (knockout cell lysate ab257097) was used. Wild-type and CD276 knockout samples were subjected to SDS-PAGE. ab227670 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-CD276 antibody [SP206] (ab227670)
    Western blot - Anti-CD276 antibody [SP206] (ab227670)
    Anti-CD276 antibody [SP206] (ab227670) at 1/400 dilution + HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate

    Predicted band size: 57 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast tissue sections labeling CD276 with ab227670 at 1/100 dilution (2.29 µg/ml). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Mainly membranous staining on the human breast, performed on a Leica Biosystems BOND™ RX instrument.
    The section was incubated with ab227670 for 10 mins at room temperature.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human lung SCCa tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human tonsil tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human testis tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human stomach adenocarcinoma tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human lung tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human lung adenocarcinoma tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human colon tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human breast tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human breast ductal carcinoma tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [SP206] (ab227670)

    Formalin-fixed, paraffin-embedded human stomach tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.

  • Anti-CD276 antibody [SP206] (ab227670)
    Anti-CD276 antibody [SP206] (ab227670)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (1)

Publishing research using ab227670? Please let us know so that we can cite the reference in this datasheet.

ab227670 has been referenced in 1 publication.

  • Zhao J  et al. B7-H3 is regulated by BRD4 and promotes TLR4 expression in pancreatic ductal adenocarcinoma. Int J Biochem Cell Biol 108:84-91 (2019). PubMed: 30664982

Customer reviews and Q&As

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Flow Cytometry abreview for Anti-CD276 antibody [SP206]

Excellent
Abreviews
Abreviews
abreview image
Application
Flow Cytometry
Sample
Mouse Cell (HM1 (mouse ovarian cancer cell line))
Permeabilization
Yes - BD Cytofix/Cytoperm Fixation/Permeabilization Solution kit Cat No. 554714
Gating Strategy
Live tumor cells gated by FSC, SSC and negatively selected with DAPI
Specification
HM1 (mouse ovarian cancer cell line)
Preparation
Cell harvesting/tissue preparation method: MEMa with 10% FBS + antibiotics
Sample buffer: PBS with 2% FBS
Fixation
BD Cytofix/Cytoperm Fixation/Permeabilization Solution kit Cat No. 554714
Read More

泰斗 宮本

Verified customer

Submitted May 21 2019

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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