BAT3/BAG6 Antibody

Western blot shows lysates of A431 human epithelial carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, BaF3 mouse pro-B cell line, CH-1 mouse B cell lymphoma cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat BAT3/BAG6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6438) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for BAT3/BAG6 at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

BAT3/BAG6 was detected in immersion fixed immature human dendritic cells using Sheep Anti-Human/Mouse/Rat BAT3/BAG6 Affinity-purified Polyclonal Antibody (Catalog # AF6438) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counter-stained with DAPI (blue). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

H2O2 induces BAG3 reduction.A Immunoblot results indicated H2O2 treatment caused time and dosage-dependent decreases in neuronal BAG3. B qPCR demonstrated H2O2 treatment causes significant decreases in neuronal BAG3 RNA. C In addition to validating H2O2-induced reductions in BAG3 protein, immunocytochemistry suggested neurons exposed to H2O2 treatment may develop minor dystrophies in their branching processes. D Immunoblotting revealed H2O2 reduced other BAG family members (BAG2, BAG5, BAG6), and exacerbated the reduction in BAG3 observed in response to Tat or Nef. ***p < 0.001. Scale bar equals 50 μM; ×40 magnification. Scale bar equals 20 μM; ×63 magnification. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33771978), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

BAT3/BAG6 Antibody Summary

• Immunogen: E. coli-derived recombinant human BAT3/BAG6
  Met1-Gln190
  Accession # P46379
• Specificity: Detects human, mouse, and rat BAT3/BAG6 in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: BAG6
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for BAT3/BAG6 Antibody

• BAG6
• BAT3
• BAT3HLA-B associated transcript-3
• BCL2-associated athanogene 6BAG-6
• D6S52E
• D6S52EHLA-B-associated transcript 3
• G3BAG family molecular chaperone regulator 6
• HLA-B associated transcript 3
• large proline-rich protein BAT3
• Protein G3
• Protein Scythe
• Scythe

Background

BAT3 (HLA-B-associated transcript 3; also BAG6, BAG family molecular chaperone regulator 6, Protein G3 and Scythe) is a multifunctional molecule that is not easily placed into known protein families. Although its predicted MW is 119 kDa, it runs anomalously in SDS-PAGE at 150-170 kDa. BAT3 is found in diverse cell types such as dendritic cells, hepatocytes, renal epithelial cells and tumor cells. It is found extracellularly where it is complexed to Hsp70, and activates the NKp30 receptor expressed on NK cells. Intracellularly, it binds to TGF-beta receptors I and II where, upon receptor ligation, it translocates to the nucleus and activates TGF-beta target genes. Human BAT3 is 1132 amino acids (aa) in length. It contains one Ub-like domain (aa 17-77), two Pro-rich regions (aa 195-273 and 394-681), an NLS (aa 1030-1052) plus a BAG domain (aa 1053-1101). BAT3 is phosphorylated on multiple Ser/Thr residues. Caspase 3 will cleave BAT3 after Asp1001, generating a 20 kDa C-terminal fragment. There are also multiple splice variants. Two of them share an absence of aa 185-190 that may be accompanied by a 36 aa insertion after Gly489. Over aa 1-190, human BAT3 shares 93% aa identity with mouse BAT3.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for BAT3/BAG6 Antibody (AF6438)(2)

Publications using AF6438

• R Chimienti, T Baccega, S Torchio, F Manenti, S Pellegrini, A Cospito, A Amabile, MT Lombardo, P Monti, V Sordi, A Lombardo, M Malnati, L Piemonti Engineering of immune checkpoints B7-H3 and CD155 enhances immune compatibility of MHC-I-/- iPSCs for beta cell replacement Cell Reports, 2022-09-27;40(13):111423. 2022-09-27 [PMID: 36170817] (ICC, Human)
• Duggan MR, Mohseni Ahooyi T, Parikh V, Khalili K Neuromodulation of BAG co-chaperones by HIV-1 viral proteins and H2O2: implications for HIV-associated neurological disorders Cell death discovery 3/26/2021 [PMID: 33771978]

Bioinformatics

• Gene Symbol: BAG6
• Uniprot:
  • P46379(Human)