Rabbit Recombinant Monoclonal B7H4 antibody. Suitable for mIHC, IP, Flow Cyt, WB, ICC/IF, IHC-P and reacts with Human, Mouse samples. Cited in 4 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
mIHC | IP | Flow Cyt | WB | ICC/IF | IHC-P | |
---|---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested | Tested |
Mouse | Expected | Expected | Expected | Not recommended | Expected | Tested |
Rat | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/50 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/100 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Negatively regulates T-cell-mediated immune response by inhibiting T-cell activation, proliferation, cytokine production and development of cytotoxicity. When expressed on the cell surface of tumor macrophages, plays an important role, together with regulatory T-cells (Treg), in the suppression of tumor-associated antigen-specific T-cell immunity. Involved in promoting epithelial cell transformation.
B7H4, UNQ659/PRO1291, VTCN1, B7H4, UNQ659/PRO1291, V-set domain-containing T-cell activation inhibitor 1, B7 homolog 4, B7h.5, Immune costimulatory protein B7-H4, Protein B7S1, T-cell costimulatory molecule B7x, B7-H4
Rabbit Recombinant Monoclonal B7H4 antibody. Suitable for mIHC, IP, Flow Cyt, WB, ICC/IF, IHC-P and reacts with Human, Mouse samples. Cited in 4 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR23665-20
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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 a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
B7-H4 also known as VTCN1 or B7x is a member of the B7 family of immunoregulatory proteins. It has an approximate molecular mass of 50 kDa. B7-H4 expression is detected on the surface of various cells including B cells T cells monocytes and some cancer cells. Its expression is often inducible; immune cells can express B7-H4 in response to specific conditions or signals in the environment.
B7-H4 acts as an immune checkpoint molecule that negatively regulates T cell immunity. It does not form a known complex with other proteins but instead interacts directly with T cells. By inhibiting T cell activation and proliferation B7-H4 contributes to the regulation of immune responses helping to maintain self-tolerance and prevent autoimmunity.
B7-H4 is involved in the regulation of immune checkpoints prominently in the pathways related to immune tolerance and suppression. It plays a critical role in the pathways that modulate tumor evasion of the immune system. B7-H4 is related to other immune checkpoint proteins such as PD-L1 which are also pivotal in immune regulation and therapy targets.
B7-H4 is connected to cancer progression and autoimmune conditions. Overexpression of B7-H4 in certain cancers such as ovarian and breast cancer associates with tumor immune evasion allowing tumors to grow unchecked by the immune system. Its expression links with proteins like CTLA-4 in maintaining immune suppression. B7-H4 modifications could serve as a potential target in cancer immunotherapy aiming to enhance the immune system's ability to combat malignancies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Flow cytometric analysis of SK-BR-3 (Human breast adenocarcinoma epithelial cell) cells labelling B7H4 with ab252438 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 647, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) ab150079) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.
B7H4 was immunoprecipitated from 0.35 mg SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab252438 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252438 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: SK-BR-3 whole cell lysate 10 ug
Lane 2: ab252438 IP in SK-BR-3 whole cell lysate 10 ug
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab252438 in SK-BR-3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 127 seconds
The bands at 65kDa represent glycosylated B7H4 (PMID: 15878339).
All lanes: Immunoprecipitation - Anti-B7H4 antibody [EPR23665-20] (ab252438)
Predicted band size: 31 kDa
Immunohistochemical analysis of paraffin-embedded mouse breast carcinoma tissue labeling B7H4 with ab252438 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).Positive staining on mouse breast cancer cells. The section was incubated with ab252438 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded mouse breast tissue labeling B7H4 with ab252438 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).Positive staining on mouse breast. The section was incubated with ab252438 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Blocking and diluting buffer and concentration: 5% NFDM/TBST
B7H4 is N-glycosylated protein. The level of B7H4 glycosylation varies between different cells and tissues. The bands at 65kDa and 45kDa represent glycosylated B7H4. (PMID: 15878339).
Negative control: HT-29 and HeLa. (PMID: 15878339).
Exposure time: 3 minutes
All lanes: Western blot - Anti-B7H4 antibody [EPR23665-20] (ab252438) at 1/1000 dilution
Lane 1: SK-BR-3 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2: MDA-MB-468 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 3: NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell), whole cell lysate at 10 µg
Lane 4: HT-29 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 5: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 31 kDa
Observed band size: 45 kDa, 65 kDa
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T+OE-612 cells labelling B7H4 with ab252438 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic and membranous staining in HEK-293T cell line transfected with HA-tagged B7H4 expression vector. Anti-HA tag antibody [EPR22819-101] ab236632 Anti-HA tag rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling B7H4 with ab252438 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on human breast carcinoma (PMID: 15756008). The section was incubated with ab252438 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling B7H4 with ab252438 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on human ovarian carcinoma (PMID: 27439899). The section was incubated with ab252438 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling B7H4 with ab252438 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on human breast (PMID: 27439899). The section was incubated with ab252438 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Fluorescence multiplex immunohistochemical analysis of the human breast (Formalin/PFA-fixed paraffin-embedded sections).
Panel A: merged staining of anti-B7H4 (ab252438, red; Opal™690), anti-CD10 (Anti-CD10 antibody [EPR22865-73] ab255609, gray; Opal™520) and anti-FABP4 (Anti-FABP4 antibody [EPR3579] ab92501, cyan; Opal™570) on human breast. Panel B: anti-B7H4 stained on glandular lumens. Panel C: anti-CD10 stained on myoepithelial cells. Panel D: anti-FABP4 stained on adipocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody.
The section was incubated in three rounds of staining: in the order of ab252438 at 1/100 dilution (4.69 μg/ml), Anti-CD10 antibody [EPR22865-73] ab255609 at 1/1000 dilution (0.615 μg/ml) and Anti-FABP4 antibody [EPR3579] ab92501 at 1/10000 dilution (0.047 μg/ml) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain.
Flow cytometry overlay histogram showing left SKBR3 positive cells and right negative HeLa stained with ab252438 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interactionfollowed by the antibody (ab252438) (1x 106 in 100μl at 10.0 μg/ml (1/50)) for 30min on ice.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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