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Receptor Activity Modifying
Proteins (RAMPs) and Calcitonin Receptor Like Receptor (CRLR)
The
calcitonin family of bioactive peptides
comprises of calcitonin,
amylin, two calcitonin-gene related peptides (CGRP1,
and CGRP2) and adrenomedullin
(ADM). Calcitonin is
32 aa peptide found in the parafollicular "C" cells of the thyroid in mammals.
It is also found in a number of non-mammals. It regulated the mineral (calcium
and phosphate) balance. Calcitonin causes hypercalcemia by acting as an
inhibitor of osteoclast induced bone resorption. CGRP
is a 37-aa peptide produced by tissue specific processing of the calcitonin
gene. Calcitonin is the major product in the thyroid, whereas CGRP is the major
product in neural tissues. CGRP is a potent cardiovascular agent. It has
structural similarity with amylin. CGRP is found in two isoforms (CGRP-I and
CGRP-II) that differs only by 3 amino acids.
Amylin is a 37-aa peptide produced in the
pancreatic beta-cell secretory granules and is co-released with insulin. Amylin
also has CGRP-like effects on bone metabolism. Amylin has specific binding sites
in the CNS and it may regulate gastric emptying and influence carbohydrate
metabolism.
Adrenomedullin (ADM) is a 52-aa hypotensive
peptide. It has structural similarity with CGRP and amylin. ADM is produced in
peripheral tissues, adrenal medulla, lung, and kidney. ADM has specific
receptors on astrocytes and it is unregulated in ischaemia. In general,
calcitonin family of peptides has N-terminal ring structures of 6-7 aa involving
a disulfide and an amidated C-terminal end.
The
calcitonin family peptides probably act through G-protein coupled membrane
receptors. The gene for calcitonin receptors has been cloned. It is homologous
to GPCRs in family "B" which typically recognizes regulatory peptides (secretin,
glucagons, VIP). Recently, a homolog of calcitonin receptor,
CRLR (calcitonin-receptor-like receptor human 461
aa; rat/mouse 463 aa) was identified. CRLR has 55% homology with calcitonin
receptor. Other members of calcitonin peptide family were candidate for CRLR but
CRLR seemed not to be a receptor for CGRP. Two related members of the family "A"
class of GPCR, RDC1 and G10D, were then identified as receptors for CGRP and
ADM, respectively. It is now shown that CRLR can function as either a CGRP
receptor or an ADM receptor, depending upon which members of a new family of
proteins called receptor activity modifying proteins
(RAMP1-3) are expressed. RAMPs1-3 contains an N-terminal signal peptide,
an extracellular N-terminus, a single transmembrane domain near the C-terminus,
and cytoplasmic C-terminus. RAMP1-3 displays 31% identity. RAMPS may be involved
in the transport of CRLR to the plasma membrane. RAMP1 (human, mouse, rat 148
aa) presents the CRLR receptor as a glycoprotein that functions as CGRP
receptor. RAMP1 is expressed in many tissues, including the uterus, bladder,
brain, pancreas, and GI tract. CRLR and RAMP1 are not co-expressed in all
tissues suggesting that their co-expression may define which cells express
functional CGRP receptors. RAMP2 (human 175 aa; rat
182 aa, and mouse 189 aa)-transported receptors are core-glycosylated and
function as ADM receptor. It is expressed in the lung, breast, immune system and
fetal tissues. RAMP3 is most abundant in the kidney
and lung.
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Items |
Antigen peptide location |
Antibody Host |
* Expected Ab Crossreactivity |
Neat Antisera Cat # 100 ul |
Aff. Pure IgG or Mab
(100 ug) |
* Control Peptide
(100 ug) |
|
RAMP1 |
H, 19 aa ~I,
H, 11 aa, ~CT |
Rb, poly |
H, R, M |
RAMP11-S |
RAMP11-A |
RAMP11-P |
|
RAMP2 |
H, 20 aa ~NT,
H, 9 aa, ~CT |
Rb, poly |
H, R, M |
RAMP21-S |
RAMP21-A |
RAMP21-P |
|
RAMP3 |
H, 13 aa ~I
H, 9 aa, ~CT |
Rb, poly |
H, R, M |
RAMP31-S |
RAMP31-A |
RAMP31-P |
|
RCP |
H, 18aa,
~NT |
Rb, poly |
H, R, M, C |
RCP11-S |
RCP11-A |
RCP11-P |
|
CRLR |
R, 20 aa
~CT |
Rb, poly |
R, M, H |
CRLR11-S |
CRLR11-A |
CRLR11-P |
|
CGRP
(ab # 1)
|
H, 19 aa,
~NT
|
Rb, poly
|
R, M, H
|
CGRP11-S
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CGRP11-A
|
CGRP11-P
|
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CGRP
(ab # 2)
|
H, 14 aa,
~I
|
Rb, poly
|
R, M, H
|
CGRP12-S
|
CGRP12-A
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CGRP12-P
|
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CGRP 1-37 aa (full length)
|
Human CGRP (1-37 aa,
oxidized); Cat # CGRP15-P5; 500 ug
Rat CGRP (1-37 aa, oxidized); Cat # CGRP16-P5; 500 ug
|
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CGRP II (1-37 aa)
|
Human CGRP II (1-37 aa,
oxidized); Cat # CGRP25-P5; 500 ug
Rat CGRP II (1-37 aa, oxidized); Cat # CGRP26-P5; 500 ug
|
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Amylin (ab # 1)
|
H, 1-37aa
|
Rb, poly
|
R, M, H
|
AMYL11-S
|
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AMYL11-P
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Human Amylin, 1-37aa
(Non-oxidized)
|
Amylin (1-37 aa, non-oxidized);
Cat # AMYL14-P; 100 ug
Biotinylated-Amylin (1-37 aa, non-oxidized), Cat #AMYL15-P-100;100ug |
|
Amylin, 1-37aa
(oxidized, Cys2-Cys7)
|
Human Amylin (1-37 aa,
oxidized); Cat # AMYL16-P5; 500 ug
Rat Amylin (1-37 aa, oxidized); Cat # AMYL17-P5; 500 ug |
|
Amylin
(ab # 2)
|
H, 13 aa, ~I &
R 13 aa, ~I
|
Rb, poly
|
R, M, H
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AMYL12-S
|
AMYL12-A
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AMYL12-P
|
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ADML
|
H, 17 aa, ~CT |
Rb, poly
|
R, M, H
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ADML11-S
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ADML11-A
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ADML11-P
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Human ADML 1-52 aa (full
length)
|
Human ADML 1-52 aa purified
(>99%), Cat # ADM15-P; 100 ug
|
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Control IgGs For use as -ve
controls in ELISA, Western, IHC. |
Control Mouse IgG (non
immune), Cat # 20008-1 (1 mg)
Control Rabbit IgG (non immune) , Cat # 20009-1 (1 mg)
|
M= Mouse; R=Rat; H=Human; Ha=Hamster; Rb=Rabbit; B=Bovine; C=Chicken; D=DOG; CT=
near C-terminus; NT=near N-terminus; I=Middle of protein; CL=Cytoplasmic loop;
EC=Extracellular domain; IC=Intracellular domain.
* Expected antibody crossreactivity information is mostly
based upon high (>70%) sequence conservation of antigenic/control peptides in
various species. When antibody crossreactivity has actually been experimentally
confirmed in various species, it will be mentioned in the appropriate data
sheets.
"Neat Antisera or antisera" are the unpurified antiserum and it is suitable for
ELISA and Western.
"Affinity pure" IgG may be more suitable for immunohistochemical (IHC)
applications and to reduce background in most immunological applications
including ELISA and Western.
"Control peptides" can not be used for Western as they are very short peptides.
They are intended for ELISA or antibody blocking studies to establish antibody
specificity.
Western blot +ve protein controls, where available, are semi-pure, pure or
recombinant proteins that are formulated in SDS-PAGE sample buffer (reduced).
They are recommended to be used for Western (load 10 ul/lane) for visualization
with antibodies.
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