Anti-ROR gamma antibody [EPR20006] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR gamma antibody [EPR20006] - BSA and Azide free (AB232516)

Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling ROR gamma with ab207082 at 1/3000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on the cortical region of mouse thymus shown at a high magnification (400×) (PMID : 14691482).

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 (ab207082).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

AbReview82622****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR gamma antibody [EPR20006] - BSA and Azide free (AB232516)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of paraformaldehyde-fixed mouse thymus tissue staining with ab232516 at 5µg/ml dilution. Secondary antibody was Alexa Fluor® 555 Anti-Rabbit IgG at 2 µg/ml. Samples were incubated with the primary antibody with TBS + 0.1% Tween, 3% BSA, 5% horse serum for 16 hours at 4°C. Blocking was done using 3% BSA, 5% horse serum for 1 hour at 20 degrees. Heat mediated antigen retrieval with 10mM Tris Base, 1mM EDTA, pH 9.2. Hematoxylin was used for counterstaining.

This image is courtesy of an anonymous Abreview

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ROR gamma antibody [EPR20006] - BSA and Azide free (AB232516)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Mouse thymocytes labeling ROR gamma with ab207082 at 1/50 dilution. Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

Note : Cell surface staining with PE conjugated anti-mouse CD8 and Alexa Fluor^® 647 conjugated anti mouse CD4 antibodies, followed by fixation with 4% PFA for 10 min and intracellular staining with ab207082. The image shows thymocytes in the CD4+CD8+ (red) or the CD4+CD8- (black, negative control) gate.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207082).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-ROR gamma antibody [EPR20006] - BSA and Azide free (AB232516)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207082).

Flow cytometry overlay histogram showing mouse thymocytes stained with ab207082 (right) and isotype control (left).

The cells were incubated in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interactions. The cells were then fixed and permeabilised with CytoFix/CytoPerm, followed by addition of the antibody (ab207082) (1x 10⁶ in 100μl at 1.0μg/ml (1/2,000)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/2000 for 30min at 22°C

Acquisition of >30,000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR gamma antibody [EPR20006] - BSA and Azide free (AB232516)

Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling ROR gamma with ab207082 at 1/3000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on mouse thymus cortex, located in the left and right of the image, with sporadic nuclear staining on medulla, middle region (PMID : 14691482). 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 (ab207082).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROR gamma antibody [EPR20006] - BSA and Azide free (AB232516)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling ROR gamma with ab207082 at 1/3000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Sporadic nuclear staining on mouse spleen (PMID : 14691482).

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 (ab207082).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-ROR gamma antibody [EPR20006] - BSA and Azide free (AB232516)

ROR gamma was immunoprecipitated from 0.35 mg of Mouse thymus lysate with ab207082 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207082 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Mouse thymus lysate, 10 μg (Input).

Lane 2 : ab207082 IP in Mouse thymus lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab207082 inMouse thymus lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207082).

All lanes:

Immunoprecipitation - Anti-ROR gamma antibody [EPR20006] (<a href='/en-us/products/primary-antibodies/ror-gamma-antibody-epr20006-ab207082'>ab207082</a>)

Predicted band size: 58 kDa

Observed band size: 58 kDa

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