Name: Anti-CD276 antibody [SP206]
Brand: RabMAb
SKU: ab227670
Rating: 5 (1 reviews)

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Western blot - 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

Carrier Free

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.

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

Alternative Versions
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)

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.

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

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) 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)

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)

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) 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)

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)

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)

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)

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)

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)

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)

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)

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)

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)

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)

Formalin-fixed, paraffin-embedded human stomach tissue stained for CD276 with ab227670 at 1/100 dilution in immunohistochemical analysis.
Anti-CD276 antibody [SP206] (ab227670)

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

Datasheet download

Download

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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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

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"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (1)

Customer reviews and Q&As

Flow Cytometry abreview for Anti-CD276 antibody [SP206]