Name: Anti-CD276 antibody [EPR20115]
Brand: RabMAb
SKU: ab219648
Rating: 4 (1 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20115] to CD276
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Human

Carrier Free

Product name

Anti-CD276 antibody [EPR20115]
See all CD276 primary antibodies
Description

Rabbit monoclonal [EPR20115] to CD276
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-Pmore details
Species reactivity

Reacts with: Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: LNCaP, U-2OS, PC-3. 293T and MDA-MB-435S whole cell lysates; Human lung, skin and placenta lysates. IHC-P: Human tonsil, placenta, hepatocellular carcinoma, lung carcinoma and breast cancer 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.

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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20115
Isotype

IgG
Research areas
- Neuroscience
- Processes

Alternative Versions
- Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human CD276 knockout HEK-293T cell line (ab266658)
KO cell lysates
- Human CD276 knockout HEK-293T cell lysate (ab257097)
Recombinant Protein
- Recombinant human CD276 protein (Fc Chimera Active) (ab216204)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab219648 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		1/1000. Detects a band of approximately 90-110 kDa (predicted molecular weight: 57 kDa).
IHC-P	(1)	1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Notes
WB 1/1000. Detects a band of approximately 90-110 kDa (predicted molecular weight: 57 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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
- Omim: 605715 Human
- SwissProt: Q5ZPR3 Human
- Unigene: 77873 Human
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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous staining on dendritic cells of human tonsil is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-CD276 antibody [EPR20115] (ab219648)

All lanes : Anti-CD276 antibody [EPR20115] (ab219648) 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 - ab219648 observed at 90-110 kDa. Red - loading control ab8245 observed at 36 kDa.

ab219648 Anti-CD276 antibody [EPR20115] 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. ab219648 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Cytoplasmic and membranous staining on vascular endothelial cells and trophoblast cells of human placenta is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-CD276 antibody [EPR20115] (ab219648)

All lanes : Anti-CD276 antibody [EPR20115] (ab219648) at 1/1000 dilution

Lane 1 : LNCaP (Human prostate cancer cell line) whole cell lysate
Lane 2 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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

Exposure time: 3 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

Raji cells were used as a negative control for CD276 (B7-H3) (PMID: 21344157).

Negative control: lane 2 Raji cell lysate
Western blot - Anti-CD276 antibody [EPR20115] (ab219648)

All lanes : Anti-CD276 antibody [EPR20115] (ab219648) at 1/2000 dilution

Lane 1 : U-2OS (Human bone osteosarcoma epithelial cell line) whole cell lysate
Lane 2 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
Lane 3 : 293T (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 4 : MDA-MB-435S (Human ductal carcinoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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

Exposure time: 3 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

CD276 (B7-H3) has a molecular weight of about 90–100 kDa, depending on the glycosylation levels. [PMID:22473715].
Western blot - Anti-CD276 antibody [EPR20115] (ab219648)

All lanes : Anti-CD276 antibody [EPR20115] (ab219648) at 1/1000 dilution

Lane 1 : Human lung lysate
Lane 2 : Human skin lysate
Lane 3 : Human placenta lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

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

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1/2: 30 seconds; Lane 3: 10 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Negative tissue: no staining on human skeletal muscle. [PMID: 18975328].

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous staining on human hepatocellular carcinoma is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous staining on human lung carcinoma is observed [PMID: 25609202].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous and cytoplasmic staining on human breast cancer tumor cells and stromal cells is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-CD276 antibody [EPR20115] (ab219648)

Western blot protocols
Immunohistochemistry protocols

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

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Publishing research using ab219648? Please let us know so that we can cite the reference in this datasheet.

ab219648 has been referenced in 1 publication.

Zhang L et al. Immune Checkpoint-Associated Locations of Diffuse Gliomas Comparing Pediatric With Adult Patients Based on Voxel-Wise Analysis. Front Immunol 12:582594 (2021). PubMed: 33815356

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Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Skin)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6.0

Permeabilization

Specification

Skin

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Oct 22 2018

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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

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD276 antibody [EPR20115]