Skip to main content

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.

Be the first to review this product! Submit a review

Images

Flow Cytometry - Anti-B7H4 antibody [EPR23665-20] (AB252438), expandable thumbnail
  • Immunoprecipitation - Anti-B7H4 antibody [EPR23665-20] (AB252438), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (AB252438), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (AB252438), expandable thumbnail
  • Western blot - Anti-B7H4 antibody [EPR23665-20] (AB252438), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
mIHCIPFlow CytWBICC/IFIHC-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

Tested
Tested

Species

Human

Dilution info

1/100

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

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.

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

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.

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

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.

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

1 product for Alternative Product

Target data

Function

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.

Alternative names

Recommended products

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.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR23665-20

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

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.

11 product images

  • Flow Cytometry - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Flow Cytometry - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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.

  • Immunoprecipitation - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Immunoprecipitation - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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

  • Western blot - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Western blot - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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

    Secondary

    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

  • Immunocytochemistry/ Immunofluorescence - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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

  • Multiplex immunohistochemistry - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Multiplex immunohistochemistry - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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 - Anti-B7H4 antibody [EPR23665-20] (ab252438), expandable thumbnail

    Flow Cytometry - Anti-B7H4 antibody [EPR23665-20] (ab252438)

    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.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com