Antibodies, Proteins, Kits and Reagents for Life Science

Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

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 - BSA and Azide free
Suitable for: IHC-P, WB
Knockout validated
Reacts with: Human

Product name

Anti-CD276 antibody [EPR20115] - BSA and Azide free
See all CD276 primary antibodies
Description

Rabbit monoclonal [EPR20115] to CD276 - BSA and Azide free
Host species

Rabbit
Tested applications

Suitable for: IHC-P, WBmore 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

Ab227577 is the carrier-free version of ab219648. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab227577 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

pH: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20115
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219648).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

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?

This data was developed using the same antibody clone in a different buffer formulation (ab219648).

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] - BSA and Azide free (ab227577)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219648).

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] - BSA and Azide free (ab227577)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219648).

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] - BSA and Azide free (ab227577)

This IHC data was generated using the same anti-CD276 antibody clone, EPR20115, in a different buffer formulation (cat# 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

This IHC data was generated using the same anti-CD276 antibody clone, EPR20115, in a different buffer formulation (cat# 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219648).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

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