Human/Mouse/Rat B7-H4 Antibody

Catalog #: MAB21542 Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB21542-100
MAB21542-SP

Detection of Human., Mouse, and Rat B7‑H4 by Western Blot.

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Human/Mouse/Rat B7-H4 Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects mouse B7-H4 in direct ELISAs. Detects human, mouse, and rat B7-H4 in Western blots.

Source

Recombinant Monoclonal Rabbit IgG Clone # 2582H

Purification

Protein A or G purified from cell culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse B7-H4
Phe29-Pro258
Accession # Q7TSP5

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Simple Western

10 µg/mL

SK‑BR‑3 human breast cancer cell line

Flow Cytometry

0.25 µg/10⁶ cells

HEK293 Human Cell Line Transfected with Mouse B7-H4 and eGFP

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

3-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human., Mouse, and Rat B7‑H4 by Western Blot. Western blot shows lysates of SK-BR-3 human breast cancer cell line, mouse uterus tissue, and rat placenta tissue. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human/Mouse/Rat B7-H4 Monoclonal Antibody (Catalog # MAB21542) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for B7-H4 at approximately 60-85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Flow Cytometry

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Detection of B7-H4 in HEK293 Human Cell Line Transfected with Mouse B7-H4 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) mouse B7-H4 or (B) irrelevant protein and eGFP was stained with Rabbit Anti-Human/Mouse B7-H4 Monoclonal Antibody (Catalog # MAB21542) followed by APC-conjugated Goat anti-Rabbit IgG Secondary Antibody (F0111). Quadrant markers were set based on Rabbit IgG control antibody staining (MAB1050). Staining was performed using our Staining Membrane-associated Proteins protocol.

Immunocytochemistry

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B7‑H4 in RAW 264.7 Mouse Cell Line. B7-H4 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Rat Anti-Mouse B7-H4 Monoclonal Antibody (Catalog # MAB21542) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western

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Detection of Human B7‑H4 by Simple Western^TM. Simple Western lane view shows lysates of SK‑BR‑3 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for B7‑H4 at approximately 128 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human/Mouse/Rat B7‑H4 Monoclonal Antibody (Catalog # MAB21542). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: B7-H4

B7-H4, also known as B7x and B7S1, belongs to the B7 family of costimulatory proteins and contains two extracellular Ig-like domains. B7-H4 is expressed on T cells, B cells, monocytes/macrophages, dendritic cells, and some tumor cells. B7-H4 signaling inhibits T cell activation, proliferation and maturation.

Long Name

B7 Homolog 4

Entrez Gene IDs

79679 (Human); 242122 (Mouse); 295322 (Rat)

Alternate Names

B7h.5; B7H4; B7-H4; B7H4T-cell costimulatory molecule B7x; B7S1; B7S1VCTN1; B7x; B7XPRO1291; FLJ22418; Immune costimulatory protein B7-H4; Protein B7S1; T cell costimulatory molecule B7x; V-set domain containing T cell activation inhibitor 1; V-set domain-containing T-cell activation inhibitor 1; Vtcn1