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B7H4 Recombinant Rabbit Monoclonal Antibody [PSH03-65] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750899

B7H4 Recombinant Rabbit Monoclonal Antibody [PSH03-65] - BSA and Azide free

Application

  • WB

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

B7H4 Recombinant Rabbit Monoclonal Antibody [PSH03-65] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human B7H4 aa 101-150 / 282.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 31 kDa

Positive Control

MDA-MB-468 cell lysate, NIH:OVCAR-3 cell lysate, mouse uterus tissue lysate, rat ovary tissue lysate, SK-Br-3.

Conjugation

unconjugated

Clone Number

PSH03-65

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:1,000

Target

Function

V-set domain-containing T-cell activation inhibitor 1 is a protein that in humans is encoded by the VTCN1 gene. B7H4 belongs to the B7 family (see CD80; MIM 112203) of costimulatory proteins. These proteins are expressed on the surface of antigen-presenting cells and interact with ligands (e.g., CD28; MIM 186760) on T lymphocytes. B7-H4 is an immune checkpoint molecule.

Background References

1. Chen L et al. The B7H4-PDL1 classifier stratifies immuno-phenotype in cervical cancer. Cancer Cell Int. 2022 Jan

2. Peuker K et al. Microbiota-dependent activation of the myeloid calcineurin-NFAT pathway inhibits B7H3- and B7H4-dependent anti-tumor immunity in colorectal cancer. Immunity. 2022 Apr

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: Q7Z7D3 Human

SwissProt: Q7TSP5 Mouse

Synonyms

B7 family member, H4 antibody

B7 H4 antibody

B7 homolog 4 antibody

B7 superfamily member 1 antibody

B7 superfamily, member 1 antibody

B7-H4 antibody

B7h.5 antibody

B7h4 antibody

B7S1 antibody

B7x antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of B7H4 on different lysates with Rabbit anti-B7H4 antibody (<a href="/products/HA750899" style="font-weight: bold;text-decoration: underline;">HA750899</a>) at 1/1,000 dilution.<br /><br />Lane 1: MDA-MB-468 cell lysate (no heat) (15 µg/Lane)<br />Lane 2: HeLa cell lysate (negative) (30 µg/Lane)<br />Lane 3: NIH:OVCAR-3 cell lysate (30 µg/Lane)<br />Lane 4: Mouse uterus tissue lysate (30 µg/Lane)<br />Lane 5: Rat ovary tissue lysate (15 µg/Lane)<br /><br />Notice: no heat means the lysate is not boiled.<br /><br />Predicted band size: 31 kDa<br />Observed band size: 60-75 kDa<br /><br />Exposure time: 35 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750899" style="font-weight: bold;text-decoration: underline;">HA750899</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of B7H4 on different lysates with Rabbit anti-B7H4 antibody (HA750899) at 1/1,000 dilution.

    Lane 1: MDA-MB-468 cell lysate (no heat) (15 µg/Lane)
    Lane 2: HeLa cell lysate (negative) (30 µg/Lane)
    Lane 3: NIH:OVCAR-3 cell lysate (30 µg/Lane)
    Lane 4: Mouse uterus tissue lysate (30 µg/Lane)
    Lane 5: Rat ovary tissue lysate (15 µg/Lane)

    Notice: no heat means the lysate is not boiled.

    Predicted band size: 31 kDa
    Observed band size: 60-75 kDa

    Exposure time: 35 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750899) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of SK-Br-3 (positive) and HeLa (negative) labeling B7H4 with Rabbit anti-B7H4 antibody (<a href="/products/HA750899" style="font-weight: bold;text-decoration: underline;">HA750899</a>) at 1/1,000 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-B7H4 antibody (<a href="/products/HA750899" style="font-weight: bold;text-decoration: underline;">HA750899</a>) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of SK-Br-3 (positive) and HeLa (negative) labeling B7H4 with Rabbit anti-B7H4 antibody (HA750899) at 1/1,000 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-B7H4 antibody (HA750899) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"