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Purified Mouse Anti-Phospholipase Cβ4
Purified Mouse Anti-Phospholipase Cβ4

Western blot analysis of Phospholipase Cβ4 expression on a rat pituitary lysate (left panel). Rat lysate was stained with Purified Mouse Anti-Phospholipase Cβ4 (Cat. No. 611540) at dilutions of 1: 1:250, 1:500, and 1:1000 (lanes 1, 2, and 3 respectively). Purified Mouse Anti-Phospholipase Cβ4 expression was visualized with HRP Goat Anti-Mouse Ig (Cat. No. 554002) and appears as a 130 kDa band.

Immunofluorescent analysis of Phospholipase Cβ4 expression on C6 cells (Rat glioma; ATCC CCL-107) (right).  Cells were seeded in a 384-well collagen coated microplate at ~ 6,000 cells per well.  After overnight incubation, cells were stained using the Triton-X 100 fix/perm protocol (see Recommended Assay Procedure) and Purified Mouse Anti-Phospholipase Cβ4.  The second step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen).  The image was taken on a BD Pathway™ 855 or 435 Bioimager using a 20x objective.  This antibody also stained SH-SY5Y (Human neuroblastoma; ATCC CRL-2266) and SK-N-SH cells (Human neuroblastoma; ATCC HTB-11) using both the Triton-X 100 and methanol fix/perm protocols (see Recommended Assay Procedure).

Western blot analysis of Phospholipase Cβ4 expression on a rat pituitary lysate (left panel). Rat lysate was stained with Purified Mouse Anti-Phospholipase Cβ4 (Cat. No. 611540) at dilutions of 1: 1:250, 1:500, and 1:1000 (lanes 1, 2, and 3 respectively). Purified Mouse Anti-Phospholipase Cβ4 expression was visualized with HRP Goat Anti-Mouse Ig (Cat. No. 554002) and appears as a 130 kDa band.

Immunofluorescent analysis of Phospholipase Cβ4 expression on C6 cells (Rat glioma; ATCC CCL-107) (right).  Cells were seeded in a 384-well collagen coated microplate at ~ 6,000 cells per well.  After overnight incubation, cells were stained using the Triton-X 100 fix/perm protocol (see Recommended Assay Procedure) and Purified Mouse Anti-Phospholipase Cβ4.  The second step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen).  The image was taken on a BD Pathway™ 855 or 435 Bioimager using a 20x objective.  This antibody also stained SH-SY5Y (Human neuroblastoma; ATCC CRL-2266) and SK-N-SH cells (Human neuroblastoma; ATCC HTB-11) using both the Triton-X 100 and methanol fix/perm protocols (see Recommended Assay Procedure).

Product Details
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BD Transduction Laboratories™
PLCβ4
Rat (QC Testing), Human, Mouse, Fly (Tested in Development)
Mouse IgG1
Human Phospholipase Cβ4 aa. 752-961
Bioimaging, Western blot (Routinely Tested), Immunofluorescence (Tested During Development)
130 kDa
250 µg/ml
AB_398998
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at -20°C.

Recommended Assay Procedures

Bioimaging:

Methanol Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample.

Triton-X 100 Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Triton is a trademark of the Dow Chemical Company.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
611540 Rev. 2
Antibody Details
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56/Phospholipase Cβ4

Phospholipase C (PLC) hydrolyzes inositol phospholipids into diacylglycerol and inositol 1,4,5-trisphosphate (IP3). Multiple distinct PLC isoenzymes have been identified and divided into three structural types: α, β, and γ. This classification is based primarily on the location of the conserved X and Y domains, whose structural integrity is essential for a functional catalytic core. The activation of PLCβ isoenzymes is uniquely regulated by G protein subunits, while PLCγ is activated following phosphorylation by protein tyrosine kinases. The β subfamily of PLC consists of at least four members: β1, β2, β3, and β4. PLCβ4 differs from the other members in that it is not activated by G protein βγ subunits, it is not found in the liver or kidney, and it is inhibited by ribonucleotides. Various isoforms of PLβC4 result from alternative splicing or proteolytic cleavage. PLCβ4 is expressed in retina and brain and knockout mice display ataxia and abnormalities in metabotropic glutamate receptor function in the cerebellum. Thus, PLCβ4 is primarily found in neuronal tissues where it is thought to be important in neurotransmitter signaling pathways.

611540 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
611540 Rev.2
Citations & References
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Development References (4)

  1. Alvarez RA, Ghalayini AJ, Xu P. cDNA sequence and gene locus of the human retinal phosphoinositide-specific phospholipase-C beta 4 (PLCB4). Genomics. 1995; 29(1):53-61. (Biology). View Reference
  2. Kano M, Hashimoto K, Watanabe M. Phospholipase cbeta4 is specifically involved in climbing fiber synapse elimination in the developing cerebellum. Proc Natl Acad Sci U S A. 1998; 95(26):15724-15729. (Biology). View Reference
  3. Kim D, Jun KS, Lee SB. Phospholipase C isozymes selectively couple to specific neurotransmitter receptors. Nature. 1997; 389(6648):290-293. (Biology). View Reference
  4. Lee CW, Park DJ, Lee KH, Kim CG, Rhee SG. Purification, molecular cloning, and sequencing of phospholipase C-beta 4. J Biol Chem. 1993; 268(28):21318-21327. (Biology). View Reference
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611540 Rev. 2

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.