Detects human Collagen IV alpha 1 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant mouse COL13A1, recombinant human (rh) COL25A1 and rhCOL23A1 is observed.
Polyclonal Sheep IgG
Mouse myeloma cell line NS0-derived recombinant human Collagen IV alpha 1 Ser1441-Thr1669 Accession # P02462
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
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.
Detection of Human Collagen IV alpha 1 by Western Blot. Western blot shows lysates of human placenta tissue. PVDF Membrane was probed with 1 µg/mL of Human Collagen IV alpha 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6308) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Collagen IV a1 at approximately 95 kDa (as indicated) ref 1. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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: Collagen IV alpha 1
COL4A1 (collagen 4-alpha 1) is a calculated 185 kDa (ref. 1) member of the type IV collagen family. It is a secreted glycoprotein that is expressed by multiple cell types, including fibroblasts, keratinocytes, and endothelial cells. Two COL4A1 molecules interact with a 170 kDa alpha 2 chain to form a collagen IV triple helix. This helix further interacts with other helices to generate covalent oligomers that form a scaffold in the basement membrane. Mature human COL4A1 is 1642 amino acids (aa) in length (SwissProt #:P02462). It has an N-terminal “7S” proregion (aa 28-172), a central collagenous domain that contains multiple Gly-based repeats (aa 173-1440), and a C‑terminal domain that is proteolytically cleaved to generate a 25-28 kDa NC1 globular segment that has potent antiangiogenic activity (aa 1441-1669). Multiple splice forms exist. One shows a deletion of aa 499-849, a second shows a seven aa substitution for aa 513-1669, and a third shows a seven aa substitution for aa 958‑1669. Over aa 1441-1669, human COL4A1 shares 97% aa sequence identity to mouse COL4A1. AF6308 detects a 95 kDa band, endogenous COL4A1 by Westen blot in human placenta tissue sample, which is consistent with the current literature (ref. 1)
1. Protein Science (2006), 15:2805-2815.
Entrez Gene IDs:
1282 (Human); 12826 (Mouse); 290905 (Rat)
alpha-1 chain; arresten; COL4A1 NC1 domain; COL4A1; collagen alpha-1(IV) chain; collagen IV, alpha-1 polypeptide; collagen, type IV, alpha 1; Collagen-4; EC 3.4.23; EC 188.8.131.52
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